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Axio Scope.A1
Operating Manual
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Zeiss Axio Scope.A1 Operating Manual

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Page 1
Operating Manual
Axio Scope.A1
Microscope for Routine and Entry-Level Research
Page 2
Carl Zeiss Copyright / Trademark Axio Scope.A1
A thorough knowledge of these instructions is mandatory in order to operate the machine. Please familiarize yourself with the content and follow the safety instructions by all means.
We reserve the right to make changes in the interest of further technical development. The manual is not subject to updating.
© Passing on and reproduction of this document, utilization and notice of its content are not
permitted unless explicitly allowed. Noncompliance is bound to indemnity.
All rights reserved, including rights created by patent grant or registration of a utility model or design.
Brands and names indicated in this manual may be trademarks of other companies. These brands and names are mentioned only for information and do not express any approval or recommendation by us.
Carl Zeiss MicroImaging GmbH cannot assume any liability for the performance or the use of these brands.
Published by: Carl Zeiss MicroImaging GmbH
PO Box 4041, D - 37030 Goettingen Telephone: +49 551 5060 660 Telefax: +49 551 5060 464 E-mail: micro@zeiss.de
www.zeiss.de
Number of the manual: M60-2-0007 e
Issue date: Version 6 – 13-05-2008
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Axio Scope.A1 Content / Figures Carl Zeiss
CONTENT
Page
1 INTRODUCTION ...................................................................................................99
1.1 Safety Guidelines............................................................................................................ 99
1.2 Warranty........................................................................................................................ 914
1.3 Stand Variants............................................................................................................... 915
1.4 Interface Diagram ......................................................................................................... 117
1.5 Summary of the Controls and Functional Elements................................................... 119
2 TECHNICAL MANUAL ........................................................................................
121
2.1 Intended Use ................................................................................................................. 121
2.2 Systems Overview ........................................................................................................ 122
2.3 Technical Data............................................................................................................... 127
3 STARTUP ............................................................................................................ 129
3.1 Mounting the Standard Components ......................................................................... 129
3.1.1 Unpacking and Mounting the Microscope Stand.............................................................. 129
3.1.2 Mounting the Upper Stand Part on the Stand Column ..................................................... 130
3.1.3 Mounting the Binocular Tube / Photo Tube...................................................................... 131
3.1.4 Inserting Eyepieces or an Auxiliary Microscope ................................................................. 131
3.1.5 Mounting Objectives........................................................................................................ 133
3.1.6 Installing and Uninstalling "Push&Click" Modules in the Reflector Insert .......................... 134
3.1.7 Mounting the Reflector Insert .......................................................................................... 135
3.1.8 Mounting the Mechanical Stage ...................................................................................... 136
3.1.9 Mechanical Stages with Friction Adjustment .................................................................... 139
3.1.10 Mounting the LED Illuminator for Transmitted Light......................................................... 141
3.1.11 Mounting the Condenser Carrier ..................................................................................... 142
3.1.12 Mounting the Condenser................................................................................................. 143
3.1.13 Mounting the Stage Carrier ............................................................................................. 144
3.1.14 Inserting the Halogen Lamp 12 V, 50 W........................................................................... 145
3.1.15 Halogen Lamp HAL 100................................................................................................... 146
3.1.16 Inserting the Adjustment Aid into the Upper Stand Part FL/HBO ....................................... 149
3.1.17 HBO 50 Illuminator .......................................................................................................... 149
3.1.18 HBO 100 Illuminator ........................................................................................................ 152
3.1.19 Illumination System Colibri and External Illumination Fixture HXP 120 .............................. 153
3.2 Connection of the Power Supply................................................................................. 154
3.2.1 Lower Stand Part for LED and HAL 50 Illumination............................................................. 154
3.2.2 Lower Stand Part for HAL 100 Illumination....................................................................... 154
3.2.3 Upper Stand Part for HAL 100/HBO, FL/HBO, FL-LED ........................................................ 155
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Carl Zeiss Content / Figures Axio Scope.A1
3.3
Switching the Microscope and its Separate Illumination Fixtures on and off......... 155
3.4 Mounting Optional Components ................................................................................ 156
3.4.1 Changing the Upper Stand Part and Inserting a Centerpiece for Larger Specimens........... 156
3.4.2 Mounting the Intermediate Plate for Analyzer Slider......................................................... 157
3.4.3 Mounting the Tube Lens Turret........................................................................................ 157
3.4.4 Mounting the Magnification Changer.............................................................................. 158
3.4.5 Changing the Filters in the Reflector Module FL P&C........................................................ 158
3.4.6 Changing the Color Splitter in the Reflector Module FL P&C ............................................ 160
3.4.7 Mounting the Polarizer D or the Filter Holder ................................................................... 161
3.4.8 Mounting and Centering the Overview Fixture................................................................. 162
3.4.9 Changing the LED Module in the Upper Stand Part FL-LED............................................... 163
3.4.10 Inserting a Modulator Disk in the Condenser 0.9/1.25 H .................................................. 164
3.4.11 Changing the Diaphragm PlasDIC .................................................................................... 165
3.4.12 Changing the Diaphragm Ph-DIC on the Achromatic-Aplanatic Condenser
0.9 H D Ph DIC ................................................................................................................
165
3.4.13 Changing the Filter in the Filter Wheel Transmitted Light ................................................. 166
3.5 Default Setting of the Microscope .............................................................................. 167
3.5.1 Adjusting the Eyepiece Distance (Distance of the Pupils) on the Binocular Tube .................. 167
3.5.2 Adjusting the Viewing Height .......................................................................................... 167
3.5.3 Correction of Vision Defects with the Eyepiece Graticules ................................................ 168
4 OPERATION ....................................................................................................... 169
4.1 Illumination and Contrasting Method ........................................................................ 169
4.1.1 Adjusting the Transmitted Light/Bright-Field According to KÖHLER .................................. 169
4.1.2 Adjusting the Transmitted Light/Dark-Field According to KÖHLER .................................... 172
4.1.3 Adjusting the Transmitted Light/Phase-Contrast............................................................... 174
4.1.4 Adjusting the Transmitted Light/Differential Interference Contrast (DIC)........................... 176
4.1.5 Adjusting PlasDIC-Contrast in Transmitted Light .............................................................. 178
4.1.6 Adjusting Transmitted Light/Polarization .......................................................................... 179
4.1.7 Adjusting the Reflected Light/Bright-Field According to KÖHLER ...................................... 183
4.1.8 Adjusting the Reflected Light/Dark-Field........................................................................... 186
4.1.9 Adjusting Reflected Light DIC and Reflected Light C-DIC.................................................. 187
4.1.10 Adjusting Reflected Light TIC ........................................................................................... 188
4.1.11 Adjusting Reflected Light Polarization – Proof of Bireflexion and Reflexion pleochroism.... 191
4.1.12 Adjusting Reflected Light Fluorescence ............................................................................ 193
4.2 Operating and Functional Elements of Optional Components ................................. 195
4.2.1 Lower Stand Part for LED Transmitted Light Illumination .................................................. 195
4.2.2 Lower Stand Part with HAL 50 Illumination ...................................................................... 196
4.2.3 Lower Stand Part for HAL 100 Illumination ...................................................................... 197
4.2.4 Stand Column Vario 380 mm or 560 mm with Gear Box ................................................. 198
4.2.5 Upper Stand Part Transmitted Light ................................................................................. 199
4.2.6 Upper Stand Part FL/HBO ................................................................................................. 199
4.2.7 Upper Stand Part FL-LED ................................................................................................ 1100
4.2.8 Upper Stand Part HAL 100/HBO..................................................................................... 1101
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Axio Scope.A1 Content / Figures Carl Zeiss
4.2.9
Upper Stand Part HAL 100/HBO DIC .............................................................................. 1101
4.2.10 Sideport ISCP T60N Left................................................................................................. 1102
4.2.11 Reflector Slider 2-fold .................................................................................................... 1103
4.2.12 Reflector Turret 4-fold or 6-fold ..................................................................................... 1103
5 MAINTENANCE, CHANGING OF FUSES AND SERVICE ................................... 1109
5.1 Maintenance................................................................................................................ 1109
5.2 Technical Service......................................................................................................... 1110
5.2.1 Check-Up ...................................................................................................................... 1110
5.2.2 Changing the Fuses ....................................................................................................... 1110
5.3 Trouble Shooting ........................................................................................................ 1111
5.4 Service.......................................................................................................................... 1114
6 ANNEX .............................................................................................................1115
6.1 List of Abbreviations .................................................................................................. 1115
6.2 Subject Index............................................................................................................... 1117
6.3 Property Rights ........................................................................................................... 1121
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Carl Zeiss Content / Figures Axio Scope.A1
FIGURES
0Fig. 1-1 Caution sign on lower stand part for HAL 100- and HAL 50-illumination.............................188H188H13
1H1HFig. 1-2 Caution sign on upper stand part FL-LED.............................................................................189H189H13
2H2HFig. 1-3 Interface diagram (upper stand part FL/HBO and lower stand part with
HAL 50 illumination) ...........................................................................................................
190H190H18
3H3HFig. 1-4 Summary of the controls and functional elements ...............................................................191H191H20
4H4HFig. 3-1 Setting up the microscope...................................................................................................192H192H29
5H5HFig. 3-2 Mounting the upper stand part on the stand column .......................................................... 193H193H30
6H6HFig. 3-3 Mounting the binocular tube ..............................................................................................194H194H31
7H7HFig. 3-4 Inserting eyepieces.............................................................................................................. 195H195H31
8H8HFig. 3-5 Inserting an eyepiece graticule ............................................................................................196H196H32
9H9HFig. 3-6 Mounting objectives............................................................................................................197H197H33
10H10HFig. 3-7 Changing the reflector module in the reflector insert ..........................................................198H198H34
11H11HFig. 3-8 Changing the reflector module in the upper stand part FL-LED............................................199H199H34
12H12HFig. 3-9 Mounting the reflector insert ..............................................................................................200H200H35
13H13HFig. 3-10 Changing the stable mechanical stage ................................................................................201H201H36
14H14HFig. 3-11 Changing the rotatable mechanical stage............................................................................202H202H36
15H15HFig. 3-12 Centering the rotatable mechanical stage ...........................................................................203H203H37
16H16HFig. 3-13 Adjusting ergonometric drive .............................................................................................. 204H204H38
17H17HFig. 3-14 Adjusting the frictional momentum.....................................................................................205H205H39
18H18HFig. 3-15 Adjusting the connecting rod position for the ergo drive..................................................... 206H206H40
19H19HFig. 3-16 Mounting the LED illuminator on the condenser..................................................................207H207H41
20H20HFig. 3-17 Connecting the LED illuminator...........................................................................................208H208H41
21H21HFig. 3-18 Mounting the condenser carrier .......................................................................................... 209H209H42
22H22HFig. 3-19 Mounting the condenser..................................................................................................... 210H210H43
23H23HFig. 3-20 Mounting the stage carrier..................................................................................................211H211H44
24H24HFig. 3-21 Removing a halogen lamp HAL 50 ......................................................................................212H212H45
25H25HFig. 3-22 Inserting a halogen lamp 12 V, 50 W .................................................................................. 213H213H45
26H26HFig. 3-23 Mounting the halogen lamp HAL 100 ................................................................................. 214H214H46
27H27HFig. 3-24 Adjusting a halogen lamp HAL 100..................................................................................... 215H215H47
28H28HFig. 3-25 Changing a halogen lamp................................................................................................... 216H216H48
29H29HFig. 3-26 Inserting the adjustment aid................................................................................................217H217H49
30H30HFig. 3-27 Opening the HBO 50...........................................................................................................218H218H49
31H31HFig. 3-28 Changing the burner...........................................................................................................219H219H50
32H32HFig. 3-29 Mounting the HBO 50.........................................................................................................220H220H51
33H33HFig. 3-30 Adjusting the HBO 50 ......................................................................................................... 221H221H51
34H34HFig. 3-31 Mounting the HBO 100.......................................................................................................222H222H52
35H35HFig. 3-32 Transformer HBO 100 W.....................................................................................................223H223H52
36H36HFig. 3-33 Adjustment aid ...................................................................................................................224H224H53
37H37HFig. 3-34 Adjusting the HBO 100 ....................................................................................................... 225H225H53
38H38HFig. 3-35 Lower stand part for HAL 50 illumination (proceed analogically for LED illumination)........... 226H226H54
39H39HFig. 3-36 Lower stand part for HAL 100 illumination ..........................................................................227H227H54
40H40HFig. 3-37 Auxiliary power supply unit HAL 100 (front and back) ......................................................... 228H228H55
41H41HFig. 3-38 Transformer HBO 100 W (front and back) .......................................................................... 229H229H55
42H42HFig. 3-39 Changing the upper stand part, inserting a centerpiece.......................................................230H230H56
43H43HFig. 3-40 Mounting the binocular tube ..............................................................................................231H231H57
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Axio Scope.A1 Content / Figures Carl Zeiss
44H44HFig. 3-41 Mounting the magnification changer ..................................................................................232H232H58
45H45HFig. 3-42 Changing the filters in the reflector module FL P&C.............................................................233H233H58
46H46HFig. 3-43 Mounting the filters and the color splitter ...........................................................................234H234H59
47H47HFig. 3-44 Opening the module ...........................................................................................................235H235H60
48H48HFig. 3-45 Changing the color filter ..................................................................................................... 236H236H60
49H49HFig. 3-46 Labeling the color splitter ....................................................................................................237H237H61
50H50HFig. 3-47 Mounting the polarizer D ....................................................................................................238H238H61
51H51HFig. 3-48 Mounting the overview fixture ............................................................................................239H239H62
52H52HFig. 3-49 Changing the LED module ..................................................................................................240H240H63
53H53HFig. 3-50 Modulator disk in condenser 0.9/1.25 H..............................................................................241H241H64
54H54HFig. 3-51 Mounting the slit-diaphragm...............................................................................................242H242H65
55H55HFig. 3-52 Changing the diaphragm Ph-DIC.........................................................................................243H243H65
56H56HFig. 3-53 Changing the filter in the filter wheel transmitted light........................................................244H244H66
57H57HFig. 3-54 Adjusting the eyepiece distance on the binocular tube ........................................................245H245H67
58H58HFig. 3-55 Adjusting the viewing height on the binocular tube ............................................................246H246H67
59H59HFig. 4-1 Microscope adjustment in transmitted light/bright-field.......................................................247H247H70
60H60HFig. 4-2 Adjusting the vertical stop on the condenser carrier.............................................................248H248H71
61H61HFig. 4-3 Adjusting the vertical stop on the focusing drive..................................................................249H249H71
62H62HFig. 4-4 Centering the dark-field diaphragm on the condenser,
achromatic-aplanatic 0.9 H D Ph DIC...................................................................................
250H250H73
63H63HFig. 4-5 Centering the annular phase diaphragm (light-colored, in the condenser) and the
phase ring (dark-colored, in the object) ...............................................................................
251H251H75
64H64HFig. 4-6 Components for the transmitted light/DIC method..............................................................252H252H77
65H65HFig. 4-7 Components for transmitted light polarization ....................................................................253H253H79
66H66HFig. 4-8 Determination of the polarization direction n
γ'
in a synthetic fiber .......................................
254H254H80
67H67HFig. 4-9 Diagram of the color tables according to Michel-Lévy.......................................................... 255H255H81
68H68HFig. 4-10 External auxiliary power supply unit for HAL 100.................................................................256H256H83
69H69HFig. 4-11 Adjusting the microscope in the reflected light/bright-field..................................................257H257H85
70H70HFig. 4-12 Compensator compartment 6x20 with C-DIC slider 6x20 ....................................................258H258H87
71H71HFig. 4-13 TIC slider 6x20 ....................................................................................................................259H259H88
72H72HFig. 4-14 Interference stripes..............................................................................................................260H260H89
73H73HFig. 4-15 Components for reflected light polarization ........................................................................261H261H92
74H74HFig. 4-16 Components for reflected light fluorescence .......................................................................262H262H94
75H75HFig. 4-17 Lower stand part for LED transmitted light illumination .......................................................263H263H95
76H76HFig. 4-18 Lower stand part with HAL 50 illumination.......................................................................... 264H264H96
77H77HFig. 4-19 Lower stand part for HAL 100 illumination ..........................................................................265H265H97
78H78HFig. 4-20 Stand column Vario.............................................................................................................266H266H98
79H79HFig. 4-21 Upper stand part transmitted light ......................................................................................267H267H99
80H80HFig. 4-22 Upper stand part FL/HBO.....................................................................................................268H268H99
81H81HFig. 4-23 Upper stand part FL-LED....................................................................................................269H269H100
82H82HFig. 4-24 Upper stand part HAL 100/HBO ........................................................................................270H270H101
83H83HFig. 4-25 Upper stand part HAL 100/HBO DIC..................................................................................271H271H101
84H84HFig. 4-26 Sideport ISCP T60N left .....................................................................................................272H272H102
85H85HFig. 4-27 Reflector slider 2-fold ........................................................................................................273H273H103
86H86HFig. 4-28 Reflector turret 6-fold .......................................................................................................274H274H103
87H87HFig. 4-29 Condenser 0.9/1.25 H with modulator disk .......................................................................275H275H104
88H88HFig. 4-30 Nosepiece with compartment for compensators................................................................ 276H276H104
89H89HFig. 4-31 Binocular ergo tube...........................................................................................................277H277H105
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Carl Zeiss Content / Figures Axio Scope.A1
90H90HFig. 4-32 Binocular photo tube ........................................................................................................278H278H106
91H91HFig. 4-33 Adjusting the viewing height on the binocular tube ..........................................................279H279H106
92H92HFig. 4-34 Mechanical stage with object holder ................................................................................. 280H280H107
93H93HFig. 4-35 Overview fixture................................................................................................................281H281H107
94H94HFig. 4-36 Diaphragm slider 14x40 mm ............................................................................................. 282H282H108
95H95HFig. 5-1 Changing the fuses on the stand.......................................................................................283H283H110
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INTRODUCTION
Axio Scope.A1 Safety Guidelines Carl Zeiss
1 INTRODUCTION
1.1 Safety Guidelines
The microscopes Axio Scope.A1 were engineered, manufactured and tested according to DIN EN 61010-1 (IEC 61010-1) and IEC 61010-2-101, safety regulations for metering, control and laboratory equipment.
The microscopes fulfill the requirements as stated in regulation 98/79/EG (In-Vitro-Diagnostika) and carry the label
.
The present manual gives information and safety warnings which must be followed by the operator.
The following warning and instruction icons are used in the manual:
CAUTION Warns the operator of a possible danger.
CAUTION Hot surface!
CAUTION UV radiation!
BE CAREFUL, LED radiation! LED class 3B, max. 30 mW, 365 - 625 nm, Avoid direct exposure to the beam. Avoid contact with the skin!
CAUTION
Pull the plug before handling the instrument!
CAUTION Possible danger for the instrument or the system.
ATTENTION Points out instructions which should be followed carefully.
Page 10
INTRODUCTION
Carl Zeiss Safety Guidelines Axio Scope.A1
10 M60-2-0007 e 05/08
The microscope Axio Scope.A1
and its genuine accessories may only be used for microscopy methods as
indicated in this manual.
The following guidelines are especially important:
The manufacturer admits no liability if the microscope, its components or single parts, are used in any different manner. Liability is also excluded for any service or repair work performed by non-authorized personnel and any rights to claims under guarantee expire.
The microscope may only be plugged into an electrical outlet equipped with a safety contact. The safety aspect must not be disabled by using an extension cord which does not have a ground wire.
Whenever it becomes apparent that any of the safety mechanisms is out of order, the microscope must be switched off and protected from any accidental use. Please contact the Zeiss Service Department or the Carl Zeiss Microscopy Service before switching the microscope on again.
The lower stand parts for LED illumination and those with integrated HAL 50 illumination are equipped with a power supply unit integrated in the stand. This unit adapts to a voltage range of 100 to 240 V ±10 %, 50 / 60 Hz, no further voltage adjustment is necessary on the microscope itself. The auxiliary power supply unit SNT 12 V DC 100 W for HAL 100 covers a voltage range from
100 to 240 V AC ±10 %, 50 to 60 Hz. It adapts automatically to the applied voltage. The transformers for the HBO 100 adapt to a voltage range from 100 to 240 V AC, 50 to 60 Hz. They adapt automatically to the applied voltage. No further voltage adjustment is necessary. However, the transformer for the HBO 50 must be adjusted according to the applied voltage (100, 110, 120, 127 or 230, 240 V). Please do not forget to adjust the power frequency accordingly (50 or 60 Hz). Before you switch on the microscope, please verify that the voltage you are about to use is suitable.
Always pull the plug before you open the microscope or change a fuse!
Please make sure that the fuses are suitable for the applied nominal current. Never use any makeshift fuses and do not short-circuit the fuse holders.
The microscopes Axio Scope.A1 do not have any special safety fixtures to protect from acid, potentially infectious, toxic, radioactive or other specimens potentially hazardous to your health. All legal requirements, especially the national regulations for the prevention of accidents must be followed.
Page 11
INTRODUCTION
Axio Scope.A1 Safety Guidelines Carl Zeiss
M60-2-0007 e 05/08 11
Gas discharge lamps, e.g. HBO 100, emit ultraviolet radiation which may cause eye and skin burns. Never look directly into the beam and avoid any direct skin exposure to the beam. Never operate the microscope without the necessary safety equipment (e.g. special damping filters or the fluorescence protection screen). Gas discharge lamps have a high internal pressure when hot. Never change the lamps while they are still hot and make sure to always use protective gloves and masks when changing them.
Microscope lamps radiate a lot of heat which could damage the heat sensitive fluorescence filters. It is therefore necessary not to remove the heat protection filter when using a fluorescence filter so that it can function properly.
Avoid any contact with the hot lamp case. Before you change a lamp, make sure to pull the plug and wait 15 minutes for the lamp to cool down.
Dirt and dust may affect the microscope’s performance. Protect it by using a dust cover when it is not in use. Always make sure that the instrument is switched off before covering it up.
Covering the ventilation slits may accumulate heat which could damage the instrument and even start a fire. Make sure to keep the ventilation slits open and never put or accidentally drop anything into them.
Only authorized personnel should be allowed to operate the instruments. The personnel must be aware of the possible danger which might occur while using the microscope. The Axio Scope.A1 is a precision tool which can easily be damaged or even destroyed when handled improperly.
Never operate the instrument in an area with potentially explosive atmosphere. Always place it on a stable and heat resistant surface. Specimens must be disposed of appropriately according to valid legal regulations and internal work instructions.
When the lamp module (e.g. Colibri) is equipped with appropriate LED modules, or when a white light source coupled by a fluid light conductor is used, ultraviolet radiation may leak which can cause eye and skin burns. Never look directly into the light and avoid any direct skin exposure to the light. Always use the appropriate safety equipment when operating the microscope.
Page 12
INTRODUCTION
Carl Zeiss Safety Guidelines Axio Scope.A1
12 M60-2-0007 e 05/08
Never look into the light beam of the illumination equipment - with or without optical instruments. Not even if you just want to look at a specimen. Irreparable eye damage may occur!
Do not hold any flammable or easily combustible materials around the light beam.
Please read the safety data sheet on Immersol 518 N
®
, Immersol 518 F® and Immersol W®
carefully.
The immersion fluid Immersol 518 N® is irritant to the skin. Avoid any contact with skin, eyes and clothes. If skin contact occurs, wash off with lots of water and soap.
If eye contact occurs, flush with water immediately for at least 5 minutes. Seek medical assistance if irritation continues.
Dispose of immersion fluid Immersol 518 N
®
appropriately: Do not allow to contaminate
surface water or get into drains.
Defective microscopes do not belong in the household garbage. Follow the appropriate legal regulations for their disposal. Specimens must also be disposed off appropriately according to valid legal regulations and internal work instructions.
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INTRODUCTION
Axio Scope.A1 Safety Guidelines Carl Zeiss
M60-2-0007 e 05/08 13
Caution signs on lower stand part for HAL 100- and HAL 50-illumination and on upper stand part FL-LED
Fig. 1-1 Caution sign on lower stand part for HAL 100- and HAL 50-illumination
Fig. 1-2 Caution sign on upper stand part FL-LED
Page 14
INTRODUCTION
Carl Zeiss Warranty Axio Scope.A1
14 M60-2-0007 e 05/08
1.2 Warranty
The manufacturer guarantees that the instrument is free of any material and workmanship defects upon delivery. If you become aware of any deficiencies, please contact us immediately and take all necessary precautions in order to avoid further damage. Upon notice of deficiencies it is up to the manufacturer’s discretion to correct the deficiencies or to deliver a defect-free instrument. Defects due to ordinary wear and tear (especially on wearing parts) and to improper handling are not covered by our warranty.
The manufacturer is not responsible for damages to the instrument which occur by maloperation, negligence or any other manipulation of the instrument, especially if the damages occur while removing or changing any parts or when using accessories from other manufacturers. This will immediately void the warranty.
No maintenance or repair work, except for the instances mentioned in the manual, may be performed on the microscopes. Only Carl Zeiss service personnel or personnel authorized especially by Carl Zeiss may perform repair work on the microscopes. If your instrument happens to malfunction please contact the Carl Zeiss Microscopy Service Department, Germany, (see p. 114) or the Carl Zeiss agency assigned to your country.
Page 15
INTRODUCTION
Axio Scope.A1 Stand Variants Carl Zeiss
M60-2-0007 e 05/08 15
1.3 Stand Variants
The shipment includes five different upper stand parts, three lower stand parts and two stand columns which can be combined according to the following chart.
Upper stand
part
Lower stand part
Upper stand part Backlight
with nosepiece 6x bright field, M27
Upper stand part FL/HBO
with nosepiece 3x bright field, 3x DIC, M27
Upper stand part FL-LED
with nosepiece 3x bright field, 3x DIC, M27
Upper stand part HAL 100/HBO
with nosepiece 6x bright/dark field, M 27
Upper stand part HAL 100/HBO
with nosepiece 6x bright/dark field, DIC, M27
Lower stand part for LED illumination
Microscope stand
"Axio Scope.A1"
LED,
6x H
430035-9200-000
Microscope stand
"Axio Scope.A1"
LED, FL/HBO,
3x H, 3x DIC
430035-9210-000
Microscope stand
"Axio Scope.A1"
LED, FL-LED,
3x H, 3x DIC
430035-9220-000
Microscope stand
"Axio Scope.A1"
LED,
HAL 100/HBO,
6x HD
430035-9080-000
Microscope stand
"Axio Scope.A1"
LED,
HAL 100/HBO,
6x HD DIC
430035-9090-000
Lower stand part for HAL 50 illumination
Microscope stand
"Axio Scope.A1"
HAL 50,
6x H
430035-9030-000
Microscope stand
"Axio Scope.A1"
HAL 50,
FL/ HBO,
3x H, 3x DIC
430035-9040-000
Microscope stand
"Axio Scope.A1"
HAL 50, FL-LED,
3x H, 3x DIC
430035-9050-000
Microscope stand
"Axio Scope.A1"
HAL 50,
HAL 100/HBO,
6x HD
430035-9100-000
Microscope stand
"Axio Scope.A1"
HAL 50,
HAL 100/HBO,
6x HD DIC
430035-9110-000
Lower stand part for HAL 100 illumination
Microscope stand
"Axio Scope.A1"
HAL 100, 6x H
430035-9130-000
Microscope stand
"Axio Scope.A1"
HAL 100, FL/HBO,
3x H, 3x DIC
430035-9060-000
Microscope stand
"Axio Scope.A1"
HAL 100, FL-LED,
3x H, 3x DIC
430035-9070-000
Microscope stand
"Axio Scope.A1"
HAL 100,
HAL 100/HBO,
6x HD
430035-9140-000
Microscope stand
"Axio Scope.A1"
HAL100,
HAL 100/HBO,
6x HD DIC
430035-9120-000
Page 16
INTRODUCTION
Carl Zeiss Stand Variants Axio Scope.A1
16 M60-2-0007 e 05/08
Upper stand
part
Lower stand part
Upper stand part Backlight
with nosepiece 6x bright field, M27
Upper stand part FL/HBO
with nosepiece 3x bright field, 3x DIC, M27
Upper stand part FL-LED
with nosepiece 3x bright field, 3x DIC, M27
Upper stand part HAL 100/HBO
with nosepiece 6x bright/dark field, M 27
Upper stand part HAL 100/HBO
with nosepiece 6x bright/dark field, DIC, M27
Stand column "Axio Scope" Vario 380 mm
Upper stand part
"Axio Scope" FL/HBO, 3x H, 3x DIC, M27
423730-9030-000
Stand column "Axio Scope" Vario 380 mm
451017-9010-00
Focusing gear box "Axio Scope" Vario, focus lift 15 mm
430036-9000-000
Upper stand part "Axio Scope" FL-LED, 3x H, 3x DIC, M27
423730-9050-000
Stand column "Axio Scope" Vario 380 mm
451017-9010-00
Focusing gear box "Axio Scope" Vario, focus lift 15 mm
430036-9000-000
Upper stand part "Axio Scope" HAL 100/HBO, 6x HD, M27
423730-9060-000
Stand column "Axio Scope" Vario 380 mm
451017-9010-000
Focusing gear box "Axio Scope" Vario, focus lift 15 mm
430036-9000-000
Upper stand part "Axio Scope" HAL 100/HBO, 6x HD DIC, M27
423730-9070-000
Stand column "Axio Scope" Vario 380 mm
451017-9010-000
Focusing gear box "Axio Scope" Vario, focus lift 15 mm
430036-9000-000
Stand column "Axio Scope" Vario 560 mm
Upper stand part
"Axio Scope" FL/HBO, 3x H, 3x DIC, M27
423730-9030-000
Stand column "Axio Scope" Vario 560 mm
451017-9000-000
Focusing gear box "Axio Scope" Vario, focus lift 15 mm
430036-9000-000
Upper stand part "Axio Scope" FL-LED, 3x H, 3x DIC, M27
423730-9050-000
Stand column "Axio Scope" Vario 560 mm
451017-9000-000
Focusing gear box "Axio Scope" Vario, focus lift 15 mm
430036-9000-000
Upper stand part "Axio Scope" HAL 100/HBO, 6x HD, M27
423730-9060-000
Stand column "Axio Scope" Vario 560 mm
451017-9000-000
Focusing gear box "Axio Scope" Vario, focus lift 15 mm
430036-9000-000
Upper stand part "Axio Scope" HAL 100/HBO, 6x HD DIC, M27
423730-9070-000
Stand column "Axio Scope" Vario 560 mm
451017-9000-000
Focusing gear box "Axio Scope" Vario, focus lift 15 mm
430036-9000-000
Page 17
INTRODUCTION
Axio Scope.A1 Interface Diagram Carl Zeiss
M60-2-0007 e 05/08 17
1.4 Interface Diagram
The following figure shows a diagram of the interfaces of the microscope stand. The combination upper stand part FL/HBO and lower stand part with HAL 50 illumination serves as demonstration.
The interfaces vary according to which upper or lower stand part has been chosen.
Legend of
284H284HFig. 1-3:
1 Tube 2 Achromatic illumination adapter 3 Upper stand part 4 Filter slider reflected light 5 Adjustment aid 6 Adapter 7 Reflector insert 8 Lower stand part 9 Filter slider transmitted light 10 Filter in the filter wheel transmitted light 11 Condenser carrier 12 Condenser 13 Stage carrier 14 Stage
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INTRODUCTION
Carl Zeiss Interface Diagram Axio Scope.A1
18 M60-2-0007 e 05/08
Fig. 1-3 Interface diagram (upper stand part FL/HBO and lower stand part with HAL 50 illumination)
Page 19
INTRODUCTION
Axio Scope.A1 Summary of the Controls and Functional Elements Carl Zeiss
M60-2-0007 e 05/08 19
1.5 Summary of the Controls and Functional Elements
Legend of 285H285HFig. 1-4:
1 Eyepieces 2 Binocular tube part 3 Field diaphragm (non removable or as slider) 4 Adjustment aid (on upper stand part FL/HBO only) 5 Aperture diaphragm (non-removable or as slider) or FL attenuator 6 Reflected light lamp 7 Filter slider reflected light 8 Transmitted light lamp 9 Filter slider transmitted light 10 Gear shift for diffusion disk (for lower stand part with HAL 100 illumination only) 11 Focusing drive – fine adjustment (both ways) 12 Focusing drive – rough adjustment (both ways) 13 Light intensity control (operable from both sides) 14 Gear knob for adjusting the mechanical stage in direction X 15 Gear knob for adjusting the mechanical stage in direction Y 16 Gear knob for adjusting the condenser vertically (both sides) 17 Filter wheel six fold (operable from both sides, not available for lower stand part for LED illumination) 18 Centering screw for condenser (both sides) 19 Field diaphragm (not available for lower stand part for LED illumination) 20 Condenser with aperture diaphragm (with optional modulator disk) 21 Nosepiece 22 Reflector insert (changeable) 23 Gear shift for diffusion disk reflected light (not available on all upper stand parts) 24 On/off switch (not available on lower stand parts for HAL 100 illumination) 25 Release tractor for vertical stop on focusing drive
Page 20
INTRODUCTION
Carl Zeiss Summary of the Controls and Functional Elements Axio Scope.A1
20 M60-2-0007 e 05/08
Fig. 1-4 Summary of the controls and functional elements
Page 21
TECHNICAL MANUAL
Axio Scope.A1 Intended Use Carl Zeiss
M60-2-0007 e 05/08 21
2 TECHNICAL MANUAL
2.1 Intended Use
The microscopes Axio Scope.A1 are universal microscopes designed especially for applications in biological and medical fields and for material studies.
Depending on the configuration of the microscope stand, they may be used with overhead or transmitted light only or with a combination of both.
The Axio Scope.A1 may be used in typical biomedical application fields such as:
− Medical exams in laboratories (research), hospitals, doctors‘ offices,
− Science and research (colleges, universities) in medical and biological fields,
− Industrial applications (pharmacology, food technology).
The Axio Scope.A1 may also be used for material studies in:
− Metallographic laboratories,
− Motor vehicle industry and
− Microsystems technology.
Depending on the accessories for each instrument the following microscopy and contrasting methods can be performed:
Transmitted light Reflected light
− Bright field (H) − Bright field (H)
− Dark field (D) − Dark field (D)
− Phase-contrast (Ph) − Differential interference contrast (DIC)
− Differential interference contrast (DIC) − C-DIC / TIC
− PlasDIC − Fluorescence
− Polarization (Pol) − Polarization (Pol)
With a special adapter the binocular phototubes can be connected either with a microscopy camera, a reflex camera or a digital/video camera for documentation purposes. An ISCP camera can be used when working with transmitted light.
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TECHNICAL MANUAL
Carl Zeiss Systems Overview Axio Scope.A1
22 M60-2-0007 e 05/08
2.2 Systems Overview
Page 23
TECHNICAL MANUAL
Axio Scope.A1 Systems Overview Carl Zeiss
M60-2-0007 e 05/08 23
Page 24
TECHNICAL MANUAL
Carl Zeiss Systems Overview Axio Scope.A1
24 M60-2-0007 e 05/08
Page 25
TECHNICAL MANUAL
Axio Scope.A1 Systems Overview Carl Zeiss
M60-2-0007 e 05/08 25
Page 26
TECHNICAL MANUAL
Carl Zeiss Systems Overview Axio Scope.A1
26 M60-2-0007 e 05/08
Page 27
TECHNICAL MANUAL
Axio Scope.A1 Technical Data Carl Zeiss
M60-2-0007 e 05/08 27
2.3 Technical Data
Dimensions (width x depth x height)
Microscope stand Axio Scope.A1 (no DL illumination) ..................... approx. 240 mm x 340 mm x 365 mm
Stand column Vario 380 mm .......................................................... approx. 460 mm x 390 mm x 465 mm
Stand column Vario 560 mm .......................................................... approx. 460 mm x 390 mm x 645 mm
Mass
Microscope stand Axio Scope.A1 (depending on variant and accessories).................... approx. 14 to 20 kg
Stand column Vario ............................................................................................................. approx. 19 kg
Environmental conditions Transport (in packaging):
Permissible environment temperature............................................................................. -40 to +70 °C
Storage:
Permissible environment temperature............................................................................. +10 to +40 °C
Permissible humidity (no condensation).................................................................. max. 75 % at 35 °C
Operation:
Permissible environment temperature.............................................................................. -10 to +40 °C
Permissible relative humidity (no condensation)...................................................... max. 75 % at 35 °C
Height of application........................................................................................................ max. 2000 m
Air pressure.......................................................................................................... 800 hPa to 1060 hPa
Pollution level......................................................................................................................................2
Operational data
Range of application........................................................................................................................indoor
Protection class......................................................................................................................................... I
Protection type .................................................................................................................................. IP 20
...............................................................................................In consideration of CSA and UL regulations
Overvoltage category............................................................................................................................... II
Radio interference suppression ................................................................... according to EN 55011 class B
Immunity......................................................................................................... according to DIN EN 61326
Voltage range.....................................................100 to 240 V ±10 %, no voltage adjustment is necessary no vo
l
Power frequency..........................................................................................................................50/60 Hz
Power input Axio Scope.A1 with internal power supply unit ......................................................... 110 VA
Power input Axio Scope.A1 with auxiliary power supply unit 12 V DC 100 W ............................... 220 VA
Transformer mbq52ac-z for HBO 50
Range of application........................................................................................................................indoor
Protection class......................................................................................................................................... I
Protection type .................................................................................................................................. IP 20
Switchable voltage range....................................................... 100, 110, 120, 127 VAC and 230, 240 VAC
Switchable power frequency................................................................................................. 50 and 60 Hz
Power input with HBO 50 in operation ..................................................................................max. 350 VA
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TECHNICAL MANUAL
Carl Zeiss Technical Data Axio Scope.A1
28 M60-2-0007 e 05/08
Transformer HBO 100 W
Protection class......................................................................................................................................... I
Protection type .................................................................................................................................. IP 20
Voltage ...................................................................................................................100 VAC ... 240 VAC
Power frequency .....................................................................................................................50 ... 60 Hz
Power input with HBO 103 in operation ........................................................................................ 155 VA
Fuses according to IEC 127
Microscope stand Axio Scope.A1 for LED illumination in transmitted light ........... 2x T 3.15 A/H, 5x20 mm
Microscope stand Axio Scope.A1 for HAL 50 illumination in transmitted light...... 2x T 3.15 A/H, 5x20 mm
Transformer mbq52ac-z for HBO 50............................................................. ..........100 V, 127 V: 2x T 4 A
..........................................................................................................................220 V - 240 V: 2x T 2.5 A
Transformer HBO 100 W ............................................................................................T 2.0 A/H, 5x20 mm
Auxiliary power supply unit 12 V DC 100 W .......................................................... 2x T 5.0 A/H, 5x20 mm
Light sources
LED illumination DL
Power input ....................................................................................................................................7 W
Halogen lamp........................................................................................................................ 12 V / 50 W
Light source control ................................................................ infinitely variable from approx. 3 to 12 V
Halogen lamp...................................................................................................................... 12 V / 100 W
Light source control ................................................................ infinitely variable from approx. 3 to 12 V
Mercury vapor short arc lamp ........................................................................................................ HBO 50
Power input for HBO 50................................................................................................................50 W
Mercury vapor short arc lamp ...............................................................................................HBO 103 W/2
Power input for HBO 103 W/2 ....................................................................................................100 W
Illumination system Colibri
Power input ..................................................................................................................................70 W
Axio Scope.A1:
Stand with manual stage focusing
Rough drive .................................................................................................... approx. 4 mm / rotation
Fine drive ......................................................... approx. 0.4 mm / rotation; approx. 4 µm mark distance
Lift range ...................................................................................................................... approx. 25 mm
Vertical stop..........................................................................................................mechanically variable
Condenser 0.9/1.25 H with optional
Modulator disk ........................................................................................for bright field, dark field and
...........................................................................................................phase-contrast 1, 2, 3 or PlasDIC
Manual objective change ..............................................................................via nosepiece, 6-fold H, M27
Manual reflector module change ........................................................................via reflector slider 2-fold,
..................................................................................................................reflector turret 4-fold or 6-fold
Page 29
STARTUP
Axio Scope.A1 Mounting the Standard Components Carl Zeiss
M60-2-0007 e 05/08 29
3 STARTUP
The customer has the choice to either set up or mount the Axio Scope.A1 himself before putting it into operation, or have the Zeiss Customer Service do so for a fee.
Please read the Safety Guidelines carefully before assembling and operating the microscope.
The assembly activities described in the following chapters are mainly based on the assembly of a microscope stand with an upper stand part and a lower stand part. The same activities apply when assembling a microscope stand with an upper stand part and the stand column Vario – special activities for the stand column Vario are described separately.
3.1 Mounting the Standard Components
3.1.1 Unpacking and Mounting the Microscope Stand
• Unpack all the components and check for
completeness according to bill of loading.
• Set up the microscope stand (
286H286HFig. 3-1/1) on a
vibration-free, plane, stable and non-flammable surface.
• We advise to keep the original packaging in
case you want to store the instrument over a longer period of time or need to send it back to the manufacturer. If you want to dispose of the packaging, please do so in an appropriate manner.
Fig. 3-1 Setting up the microscope
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Carl Zeiss Mounting the Standard Components Axio Scope.A1
30 M60-2-0007 e 05/08
3.1.2 Mounting the Upper Stand Part on the Stand Column
If you are going to use a microscope stand consisting of an upper stand part and a stand column, please begin by assembling the upper stand part.
• Remove the magnetically mounted cover (
287H287HFig. 3-2/1) of the upper stand part by inserting a ball head
screwdriver into the assembly opening in front (
288H288HFig. 3-2/3). Lift the cover.
• Using the six hexagon socket screws (
289H289HFig. 3-2/2) included in the package, screw the upper stand part
(
290H290HFig. 3-2/4) onto the gear box (291H291HFig. 3-2/5) of the stand column.
• Replace the cover.
• In order to adjust the microscope vertically, loosen both release tractors (
292H292HFig. 3-2/6) on the stand
column and adjust the height (according to the size of the specimen) with the hand wheel (
293H293HFig. 3-2/7).
Then tighten the release tractors again.
Fig. 3-2 Mounting the upper stand part on the stand column
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3.1.3 Mounting the Binocular Tube / Photo Tube
All binocular tubes listed in the systems overview (chapter
294H294H2.2) can be mounted on the microscope
stand.
• Loosen the hexagon socket screws (
295H295HFig. 3-3/3)
with the ball head screwdriver SW 3. Remove the dust cover caps (
296H296HFig. 3-3/2 and 5) from the
bottom of the tube and the dovetail ring socket on the stand.
• Hold the binocular tube (
297H297HFig. 3-3/1) or the
binocular photo tube at an angle and insert it into the stand socket with the dovetail ring (
298H298HFig. 3-3/4). Then push it into a horizontal
position. Turn the binocular tube into the desired examination position and tighten the hexagon socket screws with the ball head screw driver.
3.1.4 Inserting Eyepieces or an Auxiliary Microscope
• Remove both dust cover caps (
299H299HFig. 3-4/1 and 4)
from the binocular tube.
• Remove both eyepieces (
300H300HFig. 3-4/2) from the
box and insert them into the binocular tube to the stop.
• Instead of an eyepiece you may insert an
auxiliary microscope (
301H301HFig. 3-4/3) into one of the
binocular nozzles in order to examine aperture, phase and dark field diaphragms and to center phase and dark field diaphragms. These diaphragms can be focused with the adjustable crystalline lens.
Fig. 3-3 Mounting the binocular tube
Fig. 3-4 Inserting eyepieces
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How to insert eyepiece graticules
The eyepieces PL 10x/23 Br. foc. are designed to be used with eyepiece graticules.
The slight image shift caused by the added glass is reversed when the zero position is marked by the red dot (
302H302HFig. 3-5/R) and not the white one (303H303HFig.
3-5/W).
The eyepiece graticules (
304H304HFig. 3-5/3) are glued into
screw-in lens parts (
305H305HFig. 3-5/4) and can thus be
changed easily. Complete lens parts with glued-in graticules can be ordered from Zeiss.
How to change a lens part:
• Unscrew the lens part (
306H306HFig. 3-5/4) with the
eyepiece graticule (
307H307HFig. 3-5/3) and remove it.
Then insert the new lens part with the eyepiece graticule.
If you want to add your own eyepiece graticule to the lens part after removing it, make sure that the writing is face down before screwing the lens part back in.
How to insert eye guards
The eyepieces are furnished with protective rubber rings to protect spectacles from being scratched. These rubber rings can be replaced by eye guards.
• Pull the rubber rings (
308H308HFig. 3-5/2) from the eyepieces, then insert the eye guards
(
309H309HFig. 3-5/1) instead.
If you find the rubber rings hard to remove from the eyepiece slots, try using a blunt stick to press them out.
Fig. 3-5 Inserting an eyepiece graticule
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M60-2-0007 e 05/08 33
3.1.5 Mounting Objectives
• Lower the mechanical stage with the stage
carrier to the bottom stop or lower the stage carrier with the dovetail.
• Remove the dust protection caps (
310H310HFig. 3-6/2)
from the appropriate nosepiece openings.
• Take the objective (
311H311HFig. 3-6/3) out of its box and
mount it to the nosepiece (
312H312HFig. 3-6/1), starting
with the smallest zoom factor (set up clockwise).
• Instead of a objective, you may mount an object
marker (
313H313HFig. 3-6/4) to any position of the
nosepiece with the aid of an adapter ring W0,8/M27 (
314H314HFig. 3-6/5). Don’t forget to recap
the object marker when not in use in order to prevent it from drying out.
Always place dust covers over the positions in the nosepiece which are presently not being used.
The adapter ring W0,8/M27 must be used with the objective W0,8.
Fig. 3-6 Mounting objectives
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Carl Zeiss Mounting the Standard Components Axio Scope.A1
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3.1.6 Installing and Uninstalling "Push&Click" Modules in the Reflector Insert
To install or uninstall the modules, the reflector insert (reflector turret 4- or 6-fold, reflector slider 2-fold) must be removed from the upper stand part.
The reflector turret in the upper stand part FL-LED cannot be removed. Install or uninstall the modules from the front after removing the cover cap.
How to install a module:
• Remove the reflector insert (
315H315HFig. 3-7/1) from the
upper stand part (chapter
316H316H3.1.7) and put it aside
with the upper side facing down.
• Insert the module (
317H317HFig. 3-7/3) (with the upper
side facing down) with the aid of its mounting elements on its left and right (
318H318HFig. 3-7/4) at a
slant from the top into the lower spring clips (
319H319HFig. 3-7/2) on the reflector socket.
• Press the module against the upper spring clips of the reflector turret until it engages firmly.
When installing the module into the upper stand part FL-LED, remove the cover cap (
320H320HFig. 3-8/1) by pulling it
forward, then turn the module (
321H321HFig.
3-8/2) by 180° and insert it into the upper spring elements. Engage it firmly by pressing it down.
How to uninstall a module:
• Disengage the module from the upper spring elements by tilting it forward, then lift it off the lower spring elements and remove the module.
• After installing or uninstalling the reflector module, don’t forget to replace the reflector insert or the cover cap.
Fig. 3-7 Changing the reflector module in
the reflector insert
Fig. 3-8 Changing the reflector module in
the upper stand part FL-LED
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M60-2-0007 e 05/08 35
3.1.7 Mounting the Reflector Insert
• First remove the cover cap (
322H322HFig. 3-9/1) by
inserting the ball head screwdriver (SW 3 mm) into the slit (
323H323HFig. 3-9/2). Turn the locking screw
counter clockwise to the stop and pull the cover cap forward.
• Put the reflector insert (
324H324HFig. 3-9/3) with the
reflector modules P&C (e.g. reflector turret 6­fold) into the upper stand part until it stops, hold it and tighten the locking screw again.
When the reflector insert is removed, the reflected light path is automatically blocked so that the glare will not affect the operator.
Fig. 3-9 Mounting the reflector insert
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3.1.8 Mounting the Mechanical Stage
Depending on the type, the mechanical stages can either be stable or they can be rotated and centered. The shift range is 75 mm in X-direction and 50 mm in Y-direction. There are stable stages available with the drive to the right or the lefts. The rotatable mechanical stage has the drive on its right.
To assemble / disassemble a stage, remove the stage carrier, then mount the stage carrier with the mounted stage onto the stand again.
(1) Stable mechanical stage 75x50 R a) How to remove the stage
• Remove the four mounting screws (
325H325HFig. 3-10/5)
on the stage carrier (
326H326HFig. 3-10/6) with the aid of
the ball head screwdriver SW 3 (
327H327HFig. 3-10/4).
• Pull off the stage (
328H328HFig. 3-10/1) from the stage
carrier. This is easier if you turn the stage and its carrier over.
b) How to mount the stage
• Put the stage (
329H329HFig. 3-10/1) onto the stage carrier (330H330HFig. 3-10/6) so that the threaded holes in the bottom
of the stage (
331H331HFig. 3-10/2) are on top of the corresponding holes in the stage carrier (332H332HFig. 3-10/3).
• Insert the four mounting screws (
333H333HFig. 3-10/5) from below through the holes in the stage carrier and
screw them into the bottom of the stage.
• Align the stage in the XY-direction before tightening the screws.
(2) Rotatable mechanical stage 75x50/240° R
Rotatable stages can only be used together with the stage carrier for rotatable stages (430710-9010-000).
a) How to remove the stage
• Loosen the screw caps (
334H334HFig. 3-11/2) of the
spring box with approx. 3 turns.
• Press the stage forward against the spring pin (
335H335HFig. 3-11/1), lift the backside off the stage
carrier (
336H336HFig. 3-11/3) and remove it by lifting it
upward.
• Tighten the screw caps (
337H337HFig. 3-11/2).
Fig. 3-10 Changing the stable mechanical
stage
Fig. 3-11 Changing the rotatable mechanical
stage
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Axio Scope.A1 Mounting the Standard Components Carl Zeiss
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b) How to mount the stage
• Loosen the screw caps (
338H338HFig. 3-11/2) of the spring box with approx. 3 turns.
• Put the stage with the dovetail ring notch onto the spring pin (
339H339HFig. 3-11/1).
• Press the stage forward against the spring pin and lower its back into the stage carrier (
340H340HFig. 3-11/3).
• Tighten the screw caps (
341H341HFig. 3-11/2).
c) How to center the stage
When using objectives with a high magnification, only one chosen objective can be centered exactly at a time.
All stages are delivered precentered, i.e. when the stage is turned, the detail of the object in the focus remains in the center of the image. If the detail shifts out of the center when the stage is turned (
342H342HFig. 3-12/5), then it becomes necessary to recenter
the stage.
• Loosen the clamping screws of the stage (
343H343HFig.
3-12/4) and the screw caps of the stage carriers (
344H344HFig. 3-12/1).
• Determine the maximal shift of the object (
345H345HFig.
3-12/5, arrow point) to the cross line in the eyepiece by turning the stage.
• Shift the object detail towards the center of the cross line by turning the two centering screws on the stage carrier (
346H346HFig. 3-12/2) each with an
hexagon head screwdriver SW 1.5 (
347H347HFig. 3-12/3)
half the length of the arrow. Repeat the procedure if the object detail shifts out of the center again when turning the stage.
• After finishing the centering procedure tighten the screw caps (
348H348HFig. 3-12/1) again.
The stage can be turned by 240° within a shifting range of y ≤ 27 mm. No turning of the stage is possible outside of this range.
(3) Adjusting the drive length on the ergonomics
An axial shift of the gear knobs by a maximum of 15 mm can enhance the drive length of the X and Y shift on the mechanical stages with ergonomic drive.
Fig. 3-12 Centering the rotatable mechanical
stage
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(4) Removing and mounting the additional
sleeves
Both gear knobs for the stages with the order no. 432035-9030-000, 432035-9040-000 and 432035-9050-000 are equipped with additional sleeves for an even more sensitive adjustment of the object position. These sleeves can be removed in those cases when it is important to shift the object more quickly.
• Loosen both clamping screws (
349H349HFig. 3-13/4) on
the lower additional sleeve (
350H350HFig. 3-13/3) and
remove them by pulling them down. Then loosen both clamping screws (
351H351HFig. 3-13/2) on
the upper additional sleeves (
352H352HFig. 3-13/1) and
remove them by pulling them down as well.
• Mount the additional sleeves again on the gear knobs by proceeding in the reverse order. Tighten the clamping screws when you are done.
(5) Adjusting the friction (smoothness of operation) on both gear knobs of the ergonomics
The smoothness of ergonomics operation is factory-adjusted to a mid degree. The operator can change the friction as follows:
a) How to adjust the X-direction
• Remove the additional sleeves (
353H353HFig. 3-13/1 and 3) from the gear knobs by loosening the clamping
screws.
• Shift the X-gear knob (
354H354HFig. 3-13/5) downward and the Y-gear knob (355H355HFig. 3-13/7) upward.
• Hold the X-gear knob (
356H356HFig. 3-13/5) and turn the light colored knurled ring above it (357H357HFig. 3-13/6) to the
right (increased smoothness) or left (decreased smoothness) until you reach the desired degree.
b) How to adjust the Y-direction
• Hold the Y-gear knob (
358H358HFig. 3-13/7) and turn the light colored knurled sleeve above it (359H359HFig. 3-13/8) to
the right (increased smoothness) or the left (decreased smoothness) until you reach the desired degree.
• Replace the additional sleeves and tighten the clamping screws.
Enhance the life span of the stage by regularly removing the particles abraded from the object carriers. Avoid getting any particles in the mechanical guiding system of the X-adjustment.
Fig. 3-13 Adjusting ergonometric drive
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3.1.9 Mechanical Stages with Friction Adjustment
Drive length and friction for the X- and Y-adjustment can be adjusted individually on the standard mechanical stage (432035-9000-000) and the mechanical stage 432035-9070-000.
(1) Adjusting the drive length on the stage
drive
The drive length of the X- and Y-drive can be adjusted by an axial shift of the gear knob (
360H360HFig. 3-14/4 and 1) within a range of approx.
15 mm.
(2) Adjusting the frictional momentum of the
gear knobs for the X-/Y-adjustment of the mechanical stage
The friction momentum of the gear knobs is factory-adjusted to a mid degree which can be changed as follows if necessary:
a) X-drive
• Slide the gear knob for the X-adjustment (
361H361HFig.
3-14/4) all the way down.
• Take the enclosed adjustment pin (
362H362HFig. 3-14/5)
out of the gear knob for the Y-adjustment (
363H363HFig.
3-14/1) and stick it into one of the holes on the lower hole nut (
364H364HFig. 3-14/3).
• Hold the gear knob for the X- adjustment (
365H365HFig. 3-14/4) and turn the hole nut with the adjustment pin
clockwise (smaller friction momentum: –) or counter clockwise (larger friction momentum: +) until you reach the desired friction degree (see
366H366HFig. 3-14).
• The adjustment should not be more than one rotation.
b) Y-drive
• Slide the gear knob for the Y-adjustment (
367H367HFig. 3-14/1) all the way up.
• Stick the enclosed adjustment pin (
368H368HFig. 3-14/5) into the hole on the upper hole nut (369H369HFig. 3-14/2).
• Hold the gear knob for the Y-adjustment (
370H370HFig. 3-14/1) and turn the hole nut with the adjustment pin
clockwise (smaller friction momentum: –) or counter clockwise (larger friction momentum: +) until you reach the desired friction degree.
• The adjustment should not be more than one rotation.
• Replace the adjustment pin in the gear knob for the Y-adjustment (
371H371HFig. 3-14/1).
Fig. 3-14 Adjusting the frictional momentum
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(3) Adjusting the connecting rod position of
the ergo drive
The connecting rod position on the ergo drive of the mechanical stage 75x50 R (432035-9010) can be adjusted individually.
• Remove the stage with the stage carrier from the stand (see chapter
372H372H3.1.13) and put it aside
face down.
• Loosen both clamping screws (
373H373HFig. 3-15/2) on
the connecting rod (
374H374HFig. 3-15/1).
• Slide the connecting rod (
375H375HFig. 3-15/1) in the
groove into the desired position.
• Tighten both clamping screws (
376H376HFig. 3-15/2).
• Mount the stage carrier with the stage on the lower stand part (see chapter
377H377H3.1.13).
Fig. 3-15 Adjusting the connecting rod
position for the ergo drive
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3.1.10 Mounting the LED Illuminator for Transmitted Light
BE CAREFUL, LED radiation!
LED class 2M, do not look into the beam!
The LED illuminator can only be inserted on the lower stand part for LED illumination.
• If the condenser carrier is already mounted,
remove the stage carrier from the stand (see chapter
378H378H3.1.13).
• Screw the bolt (
379H379HFig. 3-16/2) by hand into the
threaded holes on the bottom of the condenser carrier (
380H380HFig. 3-16/6 and 7).
• Hold the LED illuminator (
381H381HFig. 3-16/5) parallel to
the bottom of the condenser carrier (
382H382HFig. 3-16/1), pin it upward onto the bolts
(
383H383HFig. 3-16/2) and lock it with the screws
(
384H384HFig. 3-16/4) on the left side of the LED
illuminator.
• Slide the slider with the diffusion disk
(
385H385HFig. 3-16/3) into the LED illuminator or, for
transmitted light polarization contrast, slide in the polarizer for LED (427708-0000-000).
• Unscrew three screws (
386H386HFig. 3-17/1) of the cover
(
387H387HFig. 3-17/2) of the foot and remove the cover.
• Plug the LED illuminator into the port on the
cover (
388H388HFig. 3-17/3).
• Place the cover on the foot, thread the cord
through the hole in the cover and tighten the three screws.
• Mount the stage carrier (see chapter
389H389H3.1.13).
The intensity of the LED reflected light can be controlled with the control wheel (operable from both sides) (
390H390HFig. 3-17/4).
Fig. 3-16 Mounting the LED illuminator on
the condenser
Fig. 3-17 Connecting the LED illuminator
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Guidelines for the use of the LED illuminator accessories:
The slider with diffusion disk 10° emits more light for examining objects with the eyepieces. The slider with diffusion disk 80° emits a more homogeneous light for examining objects with
the camera. The slider handle may serve as a manual shutter in order to block out the disturbing self-
fluorescence in the fluorescence light. You may adjust the color temperature for examining objects by inserting color temperature
filters into the slider with diffusion disk (retainer with prescrew ring). Yellow filter 1 --> changes the color temperature of the LED (about 7500 K) to about 4500 K Yellow filter 2 --> to about 5500 K Yellow filter 3 --> to about 4000 K Polarizer LED is a special slider which can be ordered for simple DL polarization procedures. A simple polarization contrast is possible with the polarizer, yet no conoscopy can be
performed.
3.1.11 Mounting the Condenser Carrier
• Adjust the guide of the condenser carrier (
391H391HFig. 3-18/3) with the gear knob (392H392HFig. 3-18/2)
until both screws (
393H393HFig. 3-18/4) become
accessible.
• Mount the condenser carrier on the stage carrier (
394H394HFig. 3-18/1), slide it firmly and straight
up to the upper stop and tighten both screws.
Fig. 3-18 Mounting the condenser carrier
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3.1.12 Mounting the Condenser
• Slide the stage carrier with the focusing drive to the upper stop.
• Open the frontal optic (if switchable) on the condenser with the lever (
395H395HFig. 3-19/7). Unscrew
both centering screws (
396H396HFig. 3-19/5) on the
condenser carrier until you cannot see the end parts anymore.
• Slide the condenser carrier (
397H397HFig. 3-19/3) with the
gear knob for vertical adjustment (
398H398HFig. 3-19/2)
all the way down.
• Insert the condenser (
399H399HFig. 3-19/8) between the
condenser carrier (
400H400HFig. 3-19/3) and the stage
carrier (
401H401HFig. 3-19/1). Adjust the pin screw on the
bottom of the condenser in direction of the slot (
402H402HFig. 3-19/6).
• Press the condenser with the dovetail ring against the spring box (
403H403HFig. 3-19/4) of the
condenser carrier until the condenser can be placed on the condenser carrier horizontally.
• Release the condenser and its pin screw will slide into the front slot (
404H404HFig. 3-19/6).
• Tighten the centering screws until they engage in the condenser’s dovetail ring.
Proceed accordingly when mounting other condensers.
Fig. 3-19 Mounting the condenser
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3.1.13 Mounting the Stage Carrier
• Screw the shoulder screw (
405H405HFig. 3-20/3) into the
appropriate opening:
− Opening with mark 0: no adapter is
mounted for specimen holder expansion (
406H406HFig. 3-20/3),
− Opening with mark 30: adapter is mounted for expanding specimen holder by 30 mm (
407H407HFig. 3-20/2),
− Opening with mark 60: adapter is mounted for expanding specimen holder by 60 mm (
408H408HFig. 3-20/1).
• Insert the stage carrier (
409H409HFig. 3-20/5) at a slight
angle (beneath the shoulder screw) first left into the guide, then press it in straight and tighten the Tommy screw (
410H410HFig. 3-20/4) slightly.
• Push the stage carrier along the guide upward until it engages in the shoulder screw. Tighten the Tommy screw.
• Check for an exact position of the guide.
Fig. 3-20 Mounting the stage carrier
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3.1.14 Inserting the Halogen Lamp 12 V, 50 W
• Switch off the microscope and pull its plug.
Wait at least 15 min for the halogen lamp to cool down.
• Remove the halogen lamp HAL 50 (
411H411HFig. 3-21/3)
from the back of the stand (
412H412HFig. 3-21/2) and put
it down with the open side facing up.
• Remove a used halogen lamp 12 V, 50 W (if
necessary) by pulling it out of the cap (
413H413HFig.
3-22/2) in an upward motion.
• Press the new lamp (
414H414HFig. 3-22/1) with its two
cap pins delicately and carefully into the lamp cap (
415H415HFig. 3-22/2). Be careful not to touch the
protective glass of the lamp. The lamp mustn’t be canted in order to avoid bending its cap pins or damaging the lamp holder.
• Place the halogen lamp HAL 50 with the
connecting pins into the back side of the microscope and press until the lamp engages securely.
• Replace the plug of the microscope.
The intensity can be controlled with the control wheel (
416H416HFig. 3-21/1).
Fig. 3-21 Removing a halogen lamp HAL 50
Fig. 3-22 Inserting a halogen lamp 12 V, 50 W
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3.1.15 Halogen Lamp HAL 100
The halogen lamp HAL 100 may serve as light source for transmitted light and reflected light procedures (except fluorescence light) as long as either the lower stand part or the upper stand part are equipped with the appropriate connection piece. When mounting the halogen lamp, no difference is made between reflected light and transmitted light procedures.
3.1.15.1 Mounting the halogen lamp HAL 100
Before you switch on the halogen lamp, please be sure to remove the tools which are included in the lamp case. The lamp generates a lot of heat which could damage these tools (see chapter
417H417H3.1.15.3).
• Remove the cap from the reflected light or
transmitted light connector.
• Insert the lamp case (
418H418HFig. 3-23/8) with the
dovetail ring into the connector (
419H419HFig. 3-23/2 and
420H420HFig. 3-23/3). Now fixate it by tightening the
clamping screw (
421H421HFig. 3-23/1 or 422H422HFig. 3-23/9) with
the ball head screwdriver SW 3.
Due to the lack of ground clearance it is important to avoid canting the lamp when inserting it into the lower stand part. Check for straight and correct fit.
• Insert the 3-pin lamp plug (
423H423HFig. 3-23/7) into the
back of the external power supply (
424H424HFig. 3-23/6).
When mounting HAL 100 to the lower stand part for HAL 100 illumination, it is necessary to use an additional control cord (
425H425HFig. 3-23/5) for the light intensity (see chapter 426H426H3.2.2). The light
intensity of HAL 100 can then be adjusted with the control wheel (
427H427HFig. 3-23/4).
The light intensity of HAL 100 can also be adjusted with the external power supply unit. In this case, the control cord needs to be removed.
Fig. 3-23 Mounting the halogen lamp
HAL 100
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3.1.15.2 Adjusting the halogen lamp HAL 100
Rough setting
• After loosening the clamping screw (
428H428HFig. 3-23/1
or
429H429HFig. 3-23/7), remove the ready-to-operate
halogen lamp (
430H430HFig. 3-24/2) from the microscope
stand.
• Switch on the auxiliary power supply unit of the
halogen lamp HAL 100 (see chapter
431H431H3.3).
• Direct the light beam towards a projecting
surface (wall) which is at least 3 m away.
Never look directly into the light emitting aperture of the lamp!
• Adjust the adjustment screw (
432H432HFig. 3-24/3) with a
ball head screwdriver SW 3 until both images of the lamp filament appear as sharp as possible on the projection surface.
• Adjust the adjustment screws (
433H433HFig. 3-24/4 and
5) so that the lamp filament of one image exactly fills the gaps of the reflected image (
434H434HFig. 3-24/1).
Fine setting
• Mount the microscope lamp on the microscope stand again and tighten the clamping screw to lock it.
• If applicable, turn off the diffusion disk transmitted light and remove the filter from the beam path.
• Focus the objective ≤ 40x on the sample and go to a free place of the object.
• Remove the eyepiece and center the lamp filament and its reflection in the eye pupil image with the
adjustment screws (
435H435HFig. 3-24/4 and 5).
• Optimize the even illumination of the pupil image with the adjustment screws (
436H436HFig. 3-24/3).
• Switch on the diffusion disk and replace the filter wheels.
Fig. 3-24 Adjusting a halogen lamp HAL 100
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3.1.15.3 Changing the halogen lamp 12 V, 100 W
CAUTION
Hot surface!
DANGER OF BURNS! Always let the lamp case cool off for at least 15 min.!
It is not necessary to remove the lamp case from the stand when replacing the halogen lamp. Do not store the included tools for the halogen lamp (
437H437HFig. 3-25/7) in the lamp case when the
halogen lamp is in operation. The replacement lamp (
438H438HFig. 3-25/8) can
remain in the lamp case.
• Switch off the auxiliary power supply unit of the
HAL 100 (see chapter
439H439H3.2.3). Remove 3-pin plug
of the HAL 100.
• Press down the release button (
440H440HFig. 3-25/3) of
halogen lamp HAL 100 (
441H441HFig. 3-25/1). Pull the
lamp carrier (
442H442HFig. 3-25/2) out completely and
put it aside.
• Press down both spring levers (
443H443HFig. 3-25/5) and
pull out the old halogen lamp (
444H444HFig. 3-25/6) in an
upward motion.
• Press down both spring levers, insert the new
lamp into the lamp cap (
445H445HFig. 3-25/4) and release
the spring levers. Use only the included special tools (
446H446HFig. 3-25/7) when holding/grabbing the
halogen lamp. Traces of skin grease on the halogen lamp can impair its life span.
• Press the spring levers down again briefly. The
lamp is now centered.
• Replace the lamp carrier and slide it in until it
engages fully.
• Replug the 3-pin plug of HAL 100.
Fig. 3-25 Changing a halogen lamp
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3.1.16 Inserting the Adjustment Aid into the Upper Stand Part FL/HBO
• Remove the cover (
447H447HFig. 3-26/1) from the
mounting aperture of the adjustment aid.
• Insert the adjustment aid (
448H448HFig. 3-26/4) and
tighten the three included screws (
449H449HFig. 3-26/3).
• Mount the cap with the opening (
450H450HFig. 3-26/2).
Make sure it locks.
• Slide in the movable connecting piece of the
adjustment aid.
3.1.17 HBO 50 Illuminator
3.1.17.1 Inserting or changing the burner
of the HBO 50 illuminator
BE CAREFUL – Danger of burns!
Before changing the burner of HBO 50 switch off the transformer and pull its plug. Let the lamp case cool off for at least 15 min. After the lamp case has cooled down, remove the HBO 50 from the upper stand part and put it aside on a plane surface.
• Loosen the clamping screw (
451H451HFig. 3-27/1) with
the ball head screwdriver SW 3.
• Press down the release button (
452H452HFig. 3-27/2) of
the HBO 50, pull out the lamp carrier (
453H453HFig.
3-27/3) and set it aside.
Fig. 3-26 Inserting the adjustment aid
Fig. 3-27 Opening the HBO 50
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• Press the spring lever (
454H454HFig. 3-28/5) on the
heatsink up and pull the heatsink (
455H455HFig. 3-28/1)
off the inserted dummy or the used burner (
456H456HFig.
3-28/2). Lay it aside carefully.
The cord on the heatsink must not be removed or damaged.
• Press down the spring lever (
457H457HFig. 3-28/4) on the
lower lamp socket and pull the dummy / burner (
458H458HFig. 3-28/2) out of the socket (459H459HFig. 3-28/3) in an
upward motion. Release the spring lever.
Dispose of the used burners according to legal regulations. Follow the manufacturer’s guidelines!
• Press the spring lever (
460H460HFig. 3-28/4) on the lower
lamp socket down again and insert the burner into the socket (
461H461HFig. 3-28/3). Do not touch the
burner’s glass bulb. Release the spring lever.
Avoid getting fingerprints on the burner’s glass parts. Remove immediately if fingerprints occur.
• Press spring lever (
462H462HFig. 3-28/5) on the heatsink
up and mount heatsink to the burner.
• Insert lamp carrier into lamp case and slide until it engages. Tighten the clamping screw (
463H463HFig.
3-27/1).
• Write down the readings of the operation hour’s meter (transformer). After 100 operation hours the burner must be replaced.
Fig. 3-28 Changing the burner
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3.1.17.2 Mounting HBO 50 illuminator
• Mount HBO 50 illuminator (
464H464HFig. 3-29/3) on the
connection piece (
465H465HFig. 3-29/1) of the upper
stand part, align it and tighten the clamping screw (
466H466HFig. 3-29/2).
• Plug the HBO 50 connection cord into the transformer. Then plug the transformer into an electrical outlet.
• Switch the HBO 50 illuminator on and off by pressing the power switch on the front of the power supply unit. The lamp ignites automatically when switched on.
3.1.17.3 Adjusting the HBO 50 illuminator
If the Axio Scope.A1 is equipped with an adjustment aid (to be ordered separately), the HBO 50 can remain on the fluorescence reflected light illuminator when adjusting the burner.
• Switch on the illuminator after mounting it fully on the transformer. The burner ignites automatically.
If the images of the light arc and the reflection overlap, the burner’s thermal load can increase. This may shorten its lifespan.
• Make sure that the FL attenuator its set to 100 % transmission.
• Pull out the adjustment aid (
467H467HFig. 3-30/4). The
adjusting aid’s display panel (
468H468HFig. 3-30/5) shows
the burner’s light arc (in a lighter color) and its reflection (slightly darker).
• With the aid of the adjustment screw (
469H469HFig. 3-30/1) adjust the burner axially to the
mirror so that both light arcs appear the same size in the display panel of the adjustment aid (see
470H470HFig. 3-30/5).
• With the aid of the adjustment screws center the light arc and its reflection vertically (
471H471HFig. 3-30/3) and
horizontally (
472H472HFig. 3-30/2) and position those in a parallel line within the adjustment circle of the display
panel (
473H473HFig. 3-30/5). Light arc and reflection should not overlap.
• Slide the adjustment aid back in after finishing the adjustment procedure.
Fig. 3-29 Mounting the HBO 50
Fig. 3-30 Adjusting the HBO 50
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3.1.18 HBO 100 Illuminator
3.1.18.1 Inserting the mercury vapor short arc lamp HBO 103 W/2
For safety reasons, the illuminator HBO 100 and the mercury vapor short arc lamp HBO 103 W/2 are packed separately.
The first step in order to put the lamp HBO 103 W/2 in operation is to insert it in the lamp house.
Please read the manual included with the unit before inserting or replacing the lamp HBO 103 W/2.
CAUTION Please use a FL attenuator (if it is not installed permanently) in order to change the transmission. Grey filters are not resistant enough on the long run.
3.1.18.2 Mounting the HBO 100 illuminator
• Remove the cover from the reflected light
connection piece (
474H474HFig. 3-31/1).
• Insert the lamp case (
475H475HFig. 3-31/3) with the
dovetail ring into the reflected light connection piece (
476H476HFig. 3-31/1) on the back of the unit.
Tighten the clamping screw (
477H477HFig. 3-31/2) with
the ball head screwdriver SW 3.
• Connect the multi-pole HBO 100 lamp plug
with the adapter (
478H478HFig. 3-32/1) of transformer
HBO 100 W. Secure with a retaining ring.
• First plug the cord into the outlet (
479H479HFig. 3-32/2)
of transformer HBO 100 W, then insert the power plug into the electrical outlet.
Fig. 3-31 Mounting the HBO 100
Fig. 3-32 Transformer HBO 100 W
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3.1.18.3 Adjusting the HBO 100 illuminator
The HBO 100 illuminator is available in two variants (manual and automatic adjustment).
The self-adjusting HBO 100 (423011-9901-000) adjusts automatically when the transformer is turned on.
Here is how to adjust the HBO 100 illuminator with manual adjustment (423010-0000-000).
If the FL attenuator is in the reflected light beam path, make sure that it is adjusted to 100 % transmission.
• Turn on HBO 100 illuminator (
480H480HFig. 3-34/1) on
the transformer HBO 100 W (
481H481HFig. 3-38/1) and
wait until it reaches its operational temperature.
• Pull adjustment aid (
482H482HFig. 3-33/1) out of the
microscope stand. The lighter colored focal point of the lamp HBO 103 W/2 and its slightly darker reflection become visible in the black glass window of the adjustment aid.
• Use the knurled button (
483H483HFig. 3-34/4) for
collector adjustment in order to focus the lighter focal point.
• Use the adjustment screws (
484H484HFig. 3-34/2 and 3)
to focus the darker focal point (reflection) in analogy to the focal point image (
485H485HFig. 3-33/2) in
the inscribing adjustment circle.
Both focal points of the HBO 103 W/2 lamp should be close to each other in the adjustment circle of the adjustment aid!
• Replace the adjustment aid in its original
position.
3.1.19 Illumination System Colibri and External Illumination Fixture HXP 120
Please refer to the separate operation manual of Colibri and HXP 120 for installing and operating these systems.
Fig. 3-33 Adjustment aid
Fig. 3-34 Adjusting the HBO 100
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3.2 Connection of the Power Supply
3.2.1 Lower Stand Part for LED and HAL 50 Illumination
• Connect the microscope’s power supply
(
486H486HFig. 3-35/2) with its power cord to an electrical
outlet. The microscope can adjust to a power and voltage range of 100 to 240 VAC, 50 ... 60 Hz. The power supply unit adjusts auto-
matically to the available voltage.
3.2.2 Lower Stand Part for HAL 100 Illumination
• Mount halogen lamp HAL 100 (
487H487HFig. 3-36/3) to
the stand (see also chapter
488H488H3.1.15.1).
• Connect the 3-pin HAL 100 plug to the
separate auxiliary power supply unit (
489H489HFig. 3-36/2).
• Connect the cable for light intensity control
(
490H490HFig. 3-36/1) with the appropriate connecting
socket on the back of the stand or on the auxiliary power supply unit (
491H491HFig. 3-36/2).
Fig. 3-35 Lower stand part for HAL 50
illumination (proceed analogically for LED illumination)
Fig. 3-36 Lower stand part for HAL 100
illumination
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3.2.3 Upper Stand Part for HAL 100/HBO, FL/HBO, FL-LED
• Mount the halogen lamp HAL 100 to the stand
(see chapter
492H492H3.1.15.1).
• Connect the 3-pin HAL 100 plug to the
separate auxiliary power supply unit (
493H493HFig. 3-37/3).
• Connect the power cord of the auxiliary power
supply unit (
494H494HFig. 3-37/4) with an electrical
outlet.
Or
• Connect the HBO 100 plug to the transformer
HBO 100 W (
495H495HFig. 3-38/2), then connect the
power supply (
496H496HFig. 3-38/3) of the transformer to
an electrical outlet.
Or
• Connect the external ac adapter of the upper
stand part FL-LED (with integrated LED­illumination) to an electrical outlet.
3.3 Switching the Microscope and its Separate Illumination Fixtures on and off
• Turn the power switch (497H497HFig. 3-35/1) of the
microscope with lower stand part for LED or HAL 50 illumination either on or off.
• Switch on the halogen lamp for transmitted
light (lower stand part for HAL 100 illumination) and reflected light (if in use) at its separate auxiliary power supply unit (
498H498HFig. 3-37/1)
accordingly (also refer to the separate operation manual of the auxiliary power supply unit).
• Switch on or off the HBO 100 lamp for
overhead fluorescence light (if in use) with the power switch of the transformer HBO 100 W (
499H499HFig. 3-38/1).
• Switch on the LED illumination on the upper
stand part FL-LED directly (
500H500HFig. 4-23/2).
Fig. 3-37 Auxiliary power supply unit
HAL 100 (front and back)
Fig. 3-38 Transformer HBO 100 W
(front and back)
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3.4 Mounting Optional Components
Make sure to unplug the stand and the external power supply units before you begin any mounting work.
After finishing working the appropriate components must be put back into functional condition (see chapters
501H501H3.1 to 502H502H3.3).
3.4.1 Changing the Upper Stand Part and Inserting a Centerpiece for Larger Specimens
• Remove the magnetically held cover (
503H503HFig.
3-39/2) of the upper stand part by inserting a screwdriver into the special front opening (
504H504HFig.
3-39/1). Then lift the cover.
• Hold the upper stand part (
505H505HFig. 3-39/4),
unscrew the six hexagon socket screws (
506H506HFig.
3-39/3) with the ball head screwdriver SW 5 and remove the upper stand part.
• If you want to mount a centerpiece (
507H507HFig.
3-39/6), then proceed by screwing the appropriate spacers (
508H508HFig. 3-39/7) with the
included key SW 8 into the lower stand part. Then mount the centerpiece 30 mm or 60 mm
• Mount the upper stand part to be mounted
(
509H509HFig. 3-39/4) onto the lower stand part or the
centerpiece, hold it and tighten the six hexagon socket screws (
510H510HFig. 3-39/3).
• Replace the cover.
• Check the stage carrier position and change it if
necessary (see chapter
511H511H3.1.13).
Fig. 3-39 Changing the upper stand part,
inserting a centerpiece
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3.4.2 Mounting the Intermediate Plate for Analyzer Slider
• Loosen the hexagon socket screw (
512H512HFig. 3-40/5)
with the ball head screwdriver SW 3 and lift off the tube (
513H513HFig. 3-40/1).
• Use the included ring tool to unscrew the tube
lens (accessible from the bottom) and keep it free of dust.
• Screw the tube lens included with the
intermediate plate into the tube.
• Insert the intermediate plate (
514H514HFig. 3-40/2) with
the dovetail ring into the stand socket (
515H515HFig. 3-40/6), adjust it, then fixate it with the
hexagon socket screw (
516H516HFig. 3-40/4).
• Insert the analyzer slider into the slider
receptacle (
517H517HFig. 3-40/3) if applicable.
• Slightly incline the binocular tube (
518H518HFig. 3-40/1)
when inserting it into the intermediate plate, adjust it and fixate it with the hexagon socket screw (
519H519HFig. 3-40/4).
3.4.3 Mounting the Tube Lens Turret
Mounting the tube lens turret is done analogically to mounting the intermediate plate for analyzer sliders. See chapter
520H520H3.4.3.
Before using the TL turret, please remove the tube lens from the tube. Only in combination with imager tubes and eyepieces SF23 (see Systems Overview).
Fig. 3-40 Mounting the binocular tube
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3.4.4 Mounting the Magnification Changer
• If necessary, remove camera with adapter (or
dust protection cover) from the camera port of the photo tube (
521H521HFig. 3-41/2).
• Mount the magnification changer (
522H522HFig. 3-41/1)
onto the camera port, adjust it and fixate it with the retainer nut (
523H523HFig. 3-41/3).
• Mount the camera with the appropriate adapter
onto the camera port of the magnification changer.
The scanning power for changing the remagnification modules can be adjusted with the screw with a white circle on the bottom of the housing.
3.4.5 Changing the Filters in the Reflector Module FL P&C
• Remove the reflector module FL P&C
(
524H524HFig. 3-42/3) from the reflector turret and put it
aside (see also chapter
525H525H3.1.6).
• Unscrew the adapter ring (
526H526HFig. 3-42/1) with the
mounting plate included with the tools (
527H527HFig. 3-42/6).
• Turn over the reflector module and let the filter
(
528H528HFig. 3-42/2 or 5) drop onto a soft surface.
• The emission filter is inserted in
529H529HFig. 3-42/2, the
excitation filter in
530H530HFig. 3-42/5. Then fixate with
the adapter rings (
531H531HFig. 3-42/1).
Emission and excitation filters can have a name and an arrow in the girth. The arrow indicates which way the filter must be mounted on the reflector module and should always point inward (see arrows in
532H532HFig. 3-42).
In order to minimize the image offset in multiple fluorescence images, the emission filter may have another mark to factor in the point angle.
Fig. 3-41 Mounting the magnification
changer
Fig. 3-42 Changing the filters in the reflector
module FL P&C
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When mounting the appropriate emission filter into the according reflector module, the mark should be aligned with the orientation groove (
533H533HFig. 3-42/4). This guarantees that the point angle
in the reflector modules acts equally and compensates or minimizes the image offset – already low in Zeiss filter sets – even further.
When mounting a filter without any orientation marks (arrow), we advise to proceed like this:
Filters with reflecting, di-electrical coating are mounted so that the reflecting coating (
534H534HFig.
3-43/6) faces outward (in relation to the reflector module) in excitation filters (
535H535HFig. 3-43/5) and
inward in emission filters (
536H536HFig. 3-43/2).
The reflecting coating (
537H537HFig. 3-43/4) of the color
splitter (
538H538HFig. 3-43/3) faces down when mounted.
The arrows (
539H539HFig. 3-43/7) mark the path of the
illumination beam or the imaging beam.
Fig. 3-43 Mounting the filters and the color
splitter
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3.4.6 Changing the Color Splitter in the Reflector Module FL P&C
Be extremely careful when mounting the filters and the color splitter in order to avoid damaging and soiling the optical components.
We advise ordering completely equipped reflector modules FL P&C, as changing the color splitter is very challenging.
When changing the color filter, please proceed as follows:
• Remove the reflector module FL P&C from the
reflector turret (see also chapter
540H540H3.1.6).
• Loosen both slit screws (
541H541HFig. 3-44/1) with a
screwdriver.
• Hold both parts of the reflector module (the
emission part (
542H542HFig. 3-44/2) and the excitation
part (
543H543HFig. 3-44/3) together, turn them against
the mounting position and put them down.
• Now tilt the excitation part, which lies on top
(
544H544HFig. 3-45/1), upward and lift the emission part
(
545H545HFig. 3-45/4) out of the holding fixtures
(
546H546HFig. 3-45/5b).
• Remove color splitter (
547H547HFig. 3-45/2) and spring
box (
548H548HFig. 3-45/3) from the bottom part of the
module.
• Remove the old color splitter and put the new
splitter with the reflecting coating facing up carefully on the spring box (
549H549HFig. 3-45/3). Put
both pieces together into the bottom part of the module. Make sure that the side latch of the spring box is in the appropriate recess of the bottom part of the module.
Fig. 3-44 Opening the module
Fig. 3-45 Changing the color filter
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The reflecting (coated) side (
550H550HFig. 3-46/3)
of the color splitter has a tapered edge (
551H551HFig. 3-46/1) or corner (552H552HFig. 3-46/2).
• Put the excitation part of the module
(
553H553HFig. 3-45/1) on the emission part (554H554HFig. 3-45/4)
– holding fixtures (
555H555HFig. 3-45/5b) and eyelets
(
556H556HFig. 3-45/5a) latch together. Hold both parts
together and turn them back into mounting position.
• Replace the slit screws and tighten them.
• Last, stick the sticker with the name of the filter
combination on the side of the module.
3.4.7 Mounting the Polarizer D or the Filter Holder
• For an easy assembly, remove the condenser
carrier (
557H557HFig. 3-47/3) fully, with the stage carrier.
• If necessary, unscrew stop bolt and bracket bolt
with overview fixture from the condenser carrier.
• Hold the polarizer or filter holder (
558H558HFig. 3-47/6)
parallel to the bottom of the condenser carrier (
559H559HFig. 3-47/3) and screw the bracket bolt
(
560H560HFig. 3-47/4) of the polarizer (561H561HFig. 3-47/6) with
the angled adjustment lever (
562H562HFig. 3-47/5) in to
the front threaded opening at the left below the condenser carrier (
563H563HFig. 3-47/3) until it stops.
• Screw the stop bolt (
564H564HFig. 3-47/2) with the
adjustment lever (
565H565HFig. 3-47/1) to the stop into
the back threaded opening of the condenser carrier (
566H566HFig. 3-47/3).
Proceed analogically when mounting the other components listed in the Systems Overview.
Fig. 3-46 Labeling the color splitter
Fig. 3-47 Mounting the polarizer D
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3.4.8 Mounting and Centering the Overview Fixture
• If necessary, remove polarizer or filter holder
from the condenser carrier.
• Hold the overview fixture (
567H567HFig. 3-48/4) parallel
to the bottom of the condenser carrier (
568H568HFig. 3-48/3) and screw the bracket bolt
(
569H569HFig. 3-48/6) of the overview fixture with the
angled adjustment lever (
570H570HFig. 3-48/7) into the
front threaded opening to the left below the condenser carrier (
571H571HFig. 3-48/3) until it stops.
• With the adjustment lever (
572H572HFig. 3-48/1) screw
the stop bolt (
573H573HFig. 3-48/2) into the back
threaded opening of the condenser barrier (
574H574HFig. 3-48/3) to the stop.
• Retract the overview fixture into the beam path.
Make sure that the fixture is securely engaged.
• Open aperture diaphragm and field diaphragm
completely.
• Adjust both adjustment screws (
575H575HFig. 3-48/5)
with the aid of two hexagon socket screwdrivers (SW 1.5) until the field of vision is well lit.
It only makes sense to mount the overview fixture on the condenser
0.9/1.25.
Fig. 3-48 Mounting the overview fixture
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3.4.9 Changing the LED Module in the Upper Stand Part FL-LED
• Lift the covering caps (
576H576HFig. 3-49/1) off the upper
stand part.
• Remove the connecting plug (
577H577HFig. 3-49/3) of the
LED module to be changed (
578H578HFig. 3-49/2) from
the corresponding slot (
579H579HFig. 3-49/4) and pull the
LED module out of its socket.
• Put the new LED module into the socket and
plug the cable into the corresponding slot. No adjustment is necessary.
• As the LED circuit is coupled to the reflector
turret, it is necessary to make sure that LED module and fluorescence filters on the corresponding reflector turret position are compatible.
For better orientation, the positions of the LED module and those in the reflector turret are numbered.
• Replace the covers.
When you insert LED modules or fluorescence filters, make sure that they match spectrally. This guarantees that the correct LED is switched on when the reflector turret is rotated.
Fig. 3-49 Changing the LED module
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3.4.10 Inserting a Modulator Disk in the Condenser 0.9/1.25 H
• Remove the condenser (
580H580HFig. 3-50/1) from the
condenser carrier (see chapter
581H581H3.1.12).
If the condenser cannot be lowered sufficiently, e.g. with mounted overview fixture, then it might become necessary to remove it with the stage carrier, then lower it to the stop and remove the condenser.
• Loosen the clamping screw (
582H582HFig. 3-50/5) of the
condenser’s dial segment (
583H583HFig. 3-50/3) with the
screwdriver (SW 3) and pull out the dial segment.
• Slide the modulator disk (
584H584HFig. 3-50/4) with its
two-pronged forked opening pointing forward into the condenser. Make sure that the disk engages in the guide on both inner sides of the condenser. The guide serves as a stop for the modulator disk. The pin of the disk’s clamping screw must slide into the orientation groove of the condenser.
• Tighten the disk’s clamping screw with the
screwdriver (SW 3).
• Replace the condenser into its carrier (see
chapter
585H585H3.1.12).
Fig. 3-50 Modulator disk in condenser
0.9/1.25 H
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3.4.11 Changing the Diaphragm PlasDIC
How to mount the slit-diaphragm (426715-9000-
000):
• Turn the Ph-diaphragm which needs to be changed into the free aperture of the modulator disk.
• Screw the modulator disk’s centering screws (
586H586HFig. 3-51/5) with the aid of a hexagonal socket
screwdriver back until they stop.
• Unscrew the adapter ring (
587H587HFig. 3-51/2) of the
Ph-diaphragm with the included tool (
588H588HFig. 3-51/1).
• Remove Ph-diaphragm (
589H589HFig. 3-51/4) and spring
(
590H590HFig. 3-51/3).
• With the aid of the tool (
591H591HFig. 3-51/1), insert the
slit-diaphragm (
592H592HFig. 3-51/7) with the cams into
the orientation grooves (
593H593HFig. 3-51/6).
• Screw in the adapter ring (
594H594HFig. 3-51/8) included
with the slit-diaphragm with the aid of the appropriate tool (
595H595HFig. 3-51/1).
3.4.12 Changing the Diaphragm Ph-DIC on
the Achromatic-Aplanatic Condenser 0.9 H D Ph DIC
The diaphragm is changed according to the directions given in chapter
596H596H3.4.10.
After inserting the DIC module, do not tighten the condenser’s centering screw again.
How to deactivate the pre-set diaphragm mechanism which automatically opens the diaphragm when phase contrast occurs:
• Remove the cover (
597H597HFig. 3-52/1). Turn the screw (598H598HFig. 3-51/5) with the hexagon socket screwdriver 1.5
mm counter-clockwise to the stop. You can now close the aperture diaphragm for contrasting procedures in DIC.
Fig. 3-51 Mounting the slit-diaphragm
Fig. 3-52 Changing the diaphragm Ph-DIC
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3.4.13 Changing the Filter in the Filter Wheel Transmitted Light
• Remove the stage carrier (see chapter
599H599H3.1.11).
• Unscrew three screws (
600H600HFig. 3-53/3) of the field
diaphragm socket (
601H601HFig. 3-53/4) and pull the
socket out of the stand foot.
• If necessary, pull the filter to be changed
(
602H602HFig. 3-53/1) out of the corresponding position
of the filter wheel (
603H603HFig. 3-53/2).
• Put the new filter into the position.
A filter position is always designed as firmly installed shutter position.
• Repeat the procedure for all filter wheel
positions.
• Put in the field diaphragm socket and screw it in.
• Mount the stage carrier (see chapter
604H604H3.1.11).
Fig. 3-53 Changing the filter in the filter
wheel transmitted light
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3.5 Default Setting of the Microscope
3.5.1 Adjusting the Eyepiece Distance (Distance of the Pupils) on the Binocular Tube
• Adjust the eyepiece distance (distance of the
pupils) according to the individual distance of the operator’s eyes (
605H605HFig. 3-54).
The distance is good as soon as the operator sees only one round image when looking through both eyepieces!
3.5.2 Adjusting the Viewing Height
• Adjust the viewing height to the individual
needs by swinging the eyepiece barrel upward (
606H606HFig. 3-55/A) or downward (607H607HFig. 3-55/B).
The binocular ergonomic (photo) tubes (425511­0000-000, 425512-0000-000, 445514-0000-000 and 425520-9050-000) are equipped with a continuous height adjustment in the range from 44 mm to 50 mm.
The binocular piece of the ergo tube 425520­9050-000 can also be pulled out horizontally by 50 mm.
Fig. 3-54 Adjusting the eyepiece distance on
the binocular tube
Fig. 3-55 Adjusting the viewing height on
the binocular tube
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3.5.3 Correction of Vision Defects with the Eyepiece Graticules
If you are going to use an eyepiece graticule you will need two adjustable graticules, e.g. PL 10x/23 Br. foc., in order to correct different vision defects of different operators.
• Focus the lens of the adjustable eyepiece on the grid in the eyepiece graticule.
• Using the focusing drive, focus the eyepiece with the graticule on a microscopy image of an object.
• When microscopy image and graticule look sharp in this eyepiece, focus the image for the other eye
with the adjustable eye lens of the other eyepiece.
Now both microscopy images and the graticule look sharp. Any adjustment of the focus should now be performed with the focusing drive only.
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4 OPERATION
4.1 Illumination and Contrasting Method
4.1.1 Adjusting the Transmitted Light/Bright-Field According to KÖHLER
(1) General principle of operation
The transmitted light/bright-field microscopy method is among all optical microscopy methods the one which is most commonly used. High-contrast or tinted samples (e.g. a blood smear) can be examined easily and quickly.
For an imaging result which is as true to the object as possible we need not only consider the so called direct bundled beams but just as well the indirect ones, i.e. the beams which diffract and scatter on the sample details. According to ABBE, the image is more true to the object when the fraction of the indirect bundled beams is as larger as possible.
The best performance of the microscope, and especially its objective, is achieved when condenser, field diaphragm and aperture diaphragm are adjusted according to the KÖHLER illumination principle. These fundamental basic rules for adjusting a microscope are explained in detail in chapter
608H608H4.1.1 (3) "Adjusting
the transmitted light/bright-field according to KÖHLER".
(2) Instrumentation transmitted light/bright-field
Every microscope (except the one with the stand column Vario) is configured to work with the transmitted light/bright-field method.
All available condensers (except special condensers like dark-field condensers) can be used for the transmitted light/bright-field method.
(3) Adjusting the transmitted light/bright-field according to KÖHLER
− The Axio Scope.A1 has been put into operation appropriately (chapter
609H609H3).
− The Axio Scope.A1 is turned on.
• Adjust image brightness with light intensity control (
610H610HFig. 4-1/2) on microscope stand.
• Put high-contrast sample into object holder of mechanical stage.
• Bring front lens on condenser (if in use) into place (in objectives ≥ 10x) and turn the gear knob for
vertical adjustment of the condenser (
611H611HFig. 4-1/3 and 612H612HFig. 4-2/2) to the top stop. Make sure the stop is
adjusted in a way to prevent the condenser from lifting out the sample (adjusting the condenser stop, chapter
613H613H4.1.1 (4)).
• On condensers with revolver/modulator disks: turn the knurled ring (
614H614HFig. 4-2/3) to position H (bright-
field).
• Bring objective 10x in place on nosepiece (
615H615HFig. 4-1/6) and focus sample with gear knob (616H616HFig. 4-1/1).
• Close field diaphragm (
617H617HFig. 4-1/5) enough to make it visible in field of view (even if blurred)
(
618H618HFig. 4-1/A).
• Lower condenser with gear knob for vertical adjustment until edge of field diaphragm appears sharp
(
619H619HFig. 4-1/B).
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• Center the field diaphragm image with both centering screws (
620H620HFig. 4-1/4) on the condenser
carrier (
621H621HFig. 4-1/C). Open the field diaphragm
enough to make the edge of the diaphragm disappear from the field of view (
622H622HFig. 4-1/D).
• In order to adjust the aperture (contrast), take an eyepiece from the tube barrel. Looking through the barrel adjust the aperture with the adjusting lever (
623H623HFig. 4-2/4) to the size of approx.
2/3 ... 4/5 of the diameter of the objective exit pupil (
624H624HFig. 4-1/E). In most cases this aperture
gives the best contrast at almost full resolution and is thus the best compromise for the human eye.
• Replace eyepiece in tube barrel.
Each objective changes the size of the object field and the objective aperture. The centering might also vary slightly from objective to objective, so that an adjustment of field diaphragm and aperture might become necessary for optimal results.
When using objectives < 10x, the front lens of the condenser (if swivel-mounted) must be folded out and the aperture opened fully. In order to achieve a better contrast, the field diaphragm can be pulled closer when working with large object fields. To do this you must reduce the opening by a certain range. Avoid closing it too much in order to guarantee a consistent illumination of the field of view.
A bright-field adjustment according to KÖHLER is not possible when using transmitted light LED.
Fig. 4-1 Microscope adjustment in
transmitted light/bright-field
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(4) Adjusting the vertical stop on the
condenser carrier
• Loosen the setscrew (
625H625HFig. 4-2/1) on the vertical
stop with the ball-head screwdriver SW 3.
• Focus the sample with the focusing drive.
• Close the field diaphragm and adjust the
condenser vertically (
626H626HFig. 4-2/2) until you get a
sharp image.
• Perform the vertical adjustment on the condenser by small increments only to avoid lifting out the sample.
• Tighten the setscrew (
627H627HFig. 4-2/1) on the vertical
stop.
(5) Adjusting the vertical stop on the
focusing drive
• Turn the clamping lever (
628H628HFig. 4-3/1) of the stop
towards the pin stop.
• Move the stage with the aid of the focusing drive (
629H629HFig. 4-3/2) to the highest position
allowable without risking colliding with object carrier or objectives.
• Now press the clamping lever down until the stop engages again.
Fig. 4-2 Adjusting the vertical stop on the
condenser carrier
Fig. 4-3 Adjusting the vertical stop on the
focusing drive
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4.1.2 Adjusting the Transmitted Light/Dark-Field According to KÖHLER
(1) General principle of operation
Uncolored biological samples, like bacteria or living cell cultures, are sometimes hard to see in the transmitted light/bright-field due to their translucence. Yet it is completely different when you examine these samples with the transmitted light/dark-field method. You basically illuminate the sample with an illumination aperture which is higher than the one of the objective you are using.
In the dark-field, only the diffracted and scattered light portions which are important for the imaging procedure get into the objective, whereas the indirect unaffected light beams are directed past the objective. Thus a resolution of fine structures is achieved which is partially below the resolution capacity of a light microscope. The fine structures now appear bright and incandescent on a dark background.
(2) Instrumentation Condenser with transmitted light stop in position D e.g.:
− Condenser 0.9/1.25 H with modulator disk H, D, Ph 1, Ph 2, Ph 3,
− Condenser, achrom.-aplan. 0.9 H D Ph DIC,
− Dark-field condenser with dry dark-field,
− Ultra condenser.
(3) Adjusting the transmitted light/dark-field
• Adjust the illumination according to KÖHLER in analogy to the transmitted light/bright-field. Instead of the 10x objective it is necessary to use a objective with the highest possible aperture. The aperture must not exceed the border aperture of the dark-field with the used condenser.
• Adjust revolver- / modulator disks to position D and bring in condenser front optic (if applicable).
• Take eyepiece out of the tube (or replace it by the auxiliary microscope) and check the centering of the
dark-field diaphragm in the objective exit pupil. If the central dark-field diaphragm D in the universal condenser lies partially outside or is not centered to the objective exit pupil and the pupil does not appear consistently dark, then it is necessary to re-adjust the centering of the dark-field diaphragm.
• If the dark-field diaphragm needs to be centered, please use both Allen screwdrivers SW 1.5 (
630H630HFig. 4-4/1 and 4) and adjust the two centering screws (631H631HFig. 4-4/2 and 3) until the objective exit pupil
appears consistently dark. Remove the Allen screwdrivers SW 1.5 from the condenser after finishing the centering procedure.
Objectives with built-in iris apertures have apertures which are too high for the transmitted light/dark-field. Stop the iris aperture down at least until it reaches the border aperture for the dark-field of the condenser in use.
Performance criterion for the dark-field method is always a back ground for the field of view which should be as dark as possible
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• Replace the eyepiece into the tube.
• Lighter areas which might still be visible in the
field of view are reduced when the dark-field condenser is adjusted correctly and delicately. You now have an approximately sharp eye diaphragm image.
• Now adjust the field diaphragm diameter to the size of the field of view.
Dark-field samples need to be kept impeccably clean, more so than samples for any other method. A fingerprint, dust or any dirt particle can have a negative effect, as they brighten the background and reduce the contrast of the object image.
Fig. 4-4 Centering the dark-field diaphragm
on the condenser, achromatic­aplanatic 0.9 H D Ph DIC
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4.1.3 Adjusting the Transmitted Light/Phase-Contrast
(1) General principle of operation
The phase-contrast method is ideal for examining thin uncolored samples, e.g. culture cells. Generally the human eye cannot detect phase differences (variations in refractive index or thickness) within the different cell components.
The phase-contrast method converts with the aid of optical modulators, such as "annular phase diaphragm and phase ring", the small phase differences into intensity and color differences which are visible to the human eye. Important for the generation of such images is the interference of different beams in the intermediate image.
With the aid of the optically defined ring channel "annular phase diaphragm and phase ring", the high­intensity direct light portions are damped and provided with a constant phase shift. The indirect light portions, however, which are diffracted by different cell particles, bypass this optical channel and their phase is affected by the difference in the sample’s refractive index and thickness.
In the intermediate image plane, the partial beams thus differently affected achieve interference and strengthen or weaken each other – depending on their phase. As a result, these interferences create image contents with intensity differences visible to the human eye.
(2) Instrumentation
− Phase-contrast objectives with the phase rings Ph 1, Ph 2 or Ph 3 for various medium numeral apertures which can also be used in the bright-field.
− Condenser with revolver / modulator disk which contains centerable ring diaphragms Ph 1, Ph 2 and Ph 3 for various medium numeral apertures.
− The phase annular diaphragm used on the condenser must be consistent with the labeling on the objective which is used, e.g. Ph 1.
(3) Adjusting the transmitted light/phase-contrast
• Put the phase-contrast objective, e.g. labeled Ph 1, into the beam path.
• Switch on the annular phase diaphragm on the condenser’s revolver disk which has the same label as
the phase-contrast objective, e.g. 1.
• In order to check the centering and the overlap of the lighter annular diaphragm (in the condenser) with the darker phase ring (in the objective), remove an eyepiece from the tube and replace it with an auxiliary microscope. Use the adjusting fixture on the auxiliary microscope to focus the annular diaphragm and the phase ring in the objective exit pupil.
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• If the overlap is not exact (
632H632HFig. 4-5/A), the
lighter annular diaphragm must be recentered with the aid of two hexagon screwdrivers SW 1.5 (
633H633HFig. 4-4/1 and 4). Adjust the two
centering screws (
634H634HFig. 4-4/2 and 3) so that you
achieve a full overlap with the darker phase ring (
635H635HFig. 4-5/B).
• Remove the auxiliary microscope from the tube and replace it with the eyepiece.
In order to increase the image contrast, an interference broadband filter, green 32 x 4, may be mounted on the field diaphragm or inserted into the color glass carrier (if available).
A complete phase contrast can only be achieved when the light-colored annular diaphragm (in the condenser) overlaps exactly with the dark-colored phase ring (in the object) in the illumination beam path (
636H636HFig. 4-5/B).
Fig. 4-5 Centering the annular phase
diaphragm (light-colored, in the condenser) and the phase ring (dark-colored, in the object)
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4.1.4 Adjusting the Transmitted Light/Differential Interference Contrast (DIC)
(1) General principle of operation
The transmitted light/DIC method allows for a high-contrast vivid display of transparent sample details.
The light is linearly polarized by a polarizer and is separated into two beams in a birefringent prism. The two beams penetrate two parts of the sample which are closely separated. They experience different path differences due to the different refractive indices or different sample thickness. Both beams are then reunited in a second double refractive prism and have after passing the analyzer the same polarization. Therefore both beams can interfere in the intermediate image and the path differences are thus
converted into intensity differences represented by a grey scale. A compensator or λ-plate may be used for a consecutive conversion of the grey scale in a color scale.
(2) Instrumentation
− Objectives equipped with DIC fixtures, e.g. EC-Plan-Neofluare
− Nosepiece with slot for DIC slider
− DIC slider, compatible with the objectives in use
− Condenser with turret disk containing DIC prisms (e.g. condenser, achromatic-aplanatic 0.9 H D Ph
DIC)
− Analyzer module ACR P&C for transmitted light in the reflector turret/slider or analyzer slider D/A fixated or rotatable in connection with a mounted intermediate plate for analyzer slider 12x46
− Rotatable mechanical stage, if necessary
(3) Adjusting transmitted light/DIC
• Put the DIC compatible objective (
637H637HFig. 4-6/3) in place on the nosepiece. Slide the according DIC slider
(
638H638HFig. 4-6/1) into the slit of the appropriate objective position. Make sure that the slider engages firmly.
• Swing in the analyzer module (
639H639HFig. 4-6/6) on the reflector turret (640H640HFig. 4-6/7) (or slide the analyzer slider
into the intermediate plate for analyzer sliders).
• Swing in the DIC prism (position DIC on the condenser (
641H641HFig. 4-6/4).
• Adjust field diaphragm and aperture diaphragm according to the KÖHLER method.
• Adjust the optimal contrast on the DIC slider (
642H642HFig. 4-6/1) with the knurled screw (643H643HFig. 4-6/2).
Symmetrical adjustment of the DIC slider along its middle position lets the sample details appear as if they were elevated or deepened.
• If desired, the compensator λ (
644H644HFig. 4-6/5) can be put into the opening above the nosepiece in order to
create a chromatic DIC contrast.
The DIC method functions with polarized light. It is disturbed when birefringent elements, e.g. foils, are put between polarizer and analyzer, as is sometimes done when doing a histological incision. The same situation occurs with Petri dishes or specimen holders which have a plastic base. In these cases we advise using PlasDIC.
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1 DIC slider 2 Knurled screw 3 Objective on the nosepiece 4 Condenser with DIC prism 5 Compensator λ 6 Analyzer module 7 Reflector turret
Fig. 4-6 Components for the transmitted light/DIC method
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4.1.5 Adjusting PlasDIC-Contrast in Transmitted Light
(1) General principle of operation
PlasDIC can be used independently from the material of the specimen holder.
The contrast method gives a relief-like image and is especially well suited for thicker objects. The contrast is adjustable. It is possible to contrast the cavities of microtiter plates up to the edge. It is not necessary to use cultivation holders with a glass base.
(2) Instrumentation
− Abbe condenser with modulator disk and objective-dependent slit diaphragm 2 mm (A-Plan 10x
and LD A-Plan 20x) or slit diaphragm 4.5 mm (in all other cases)
− Objectives:
A-Plan 10x, 20x, 40x; LD A-Plan 20x, 32x, 40x; LD Plan-Neofluar 20x, 40x, 63x
− Slider PlasDIC depending on the objective
− Analyzer module ACR P&C for transmitted light and reflector turret /slider or analyzer slider D/A
fixated or rotatable in connection with mounted intermediate plate for analyzer slider 12x46
(3) Adjusting PlasDIC
• Fully open the aperture on the condenser.
• Put down the sample.
• Swing into the beam path the position with the slit diaphragm 2 or 4.5 mm for PlasDIC of the
condenser. Increase brightness when switching from bright-field to PlasDIC.
• Swing analyzer module into the beam path or slide analyzer slider in beam path in intermediate plate.
• Swing in the objective for PlasDIC.
• Slide the slider PlasDIC on the nosepiece into the DIC slit of the used objective.
• Adjust the contrast with the knurled screw of the slider PlasDIC. You can now see the structures in
relief or in pseudo dark-field. The best result is achieved in the relief mode.
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4.1.6 Adjusting Transmitted Light/Polarization
4.1.6.1 How to show birefringence
(1) Application
The transmitted light/polarization method is used for samples which change the polarization of the light. Such samples are called birefringent. Examples are crystals, minerals or polymers. If such birefringent substances are observed between crossed polarizers, the birefringent portion of the sample appears bright while it’s surrounding remains dark.
A birefringent substance can be recognized by turning the sample by 360° between crossed polarizers. The sample should show four bright and four dark appearances during the turning procedure. During the turning procedure, interference colors appear that range from grey (mostly for biological samples) through white, yellow and red until blue, depending on birefringence, thickness as well as sample orientation. The interference colors may be of first or higher order.
(2) Instrumentation
− Tension-free objectives
− Rotary stage Pol
− Polarizer D (rotatable or fixed), if no polarizer
is already integrated in the condenser.
− Analyzer module ACR P&C for transmitted light in the reflector turret/slider or analyzer slider D fixed or with Lambda-plate.
(3) Adjusting the microscope
• Adjust the microscope as described in chapter
645H645H4.1.1 (3) for transmitted light/bright-field
according to KÖHLER.
• Center the rotary stage Pol (
646H646HFig. 4-7/1).
• Swing the polarizer (
647H647HFig. 4-7/3) into the beam
path and position it to 0° if you are using a rotatable polarizer.
• Put analyzer slider (
648H648HFig. 4-7/2) into the slit for the compensator or swing analyzer module in on the
reflector turret/slider. Because of the crossed polarizers the field of view now appears dark.
If an analyzer slider is going to be used for working with the Axio Scope.A1, either the reflector module H P&C or an empty position must be swung in on the reflector turret/slider.
Fig. 4-7 Components for transmitted light
polarization
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• Bring the object to be examined into the field of view and turn it with the rotary stage. Normally, birefringent (anisotropic) objects will now show the same color and intensity variations as described above during rotation between crossed polarizers. Optically anotropic substances may remain dark when an isotropic direction, e.g. from optically single-axle or double-axle crystals, is put parallel to the observation direction.
4.1.6.2 Determination of the Polarization Direction n
γ '
(1) Application
The determination of the polarization direction of n
γ
or nγ' respectively (polarization direction with the
absolute or relative largest index of refraction) and n
α
or nα' respectively (polarization direction with the
absolute or relative smallest index of refraction) relative to the morphological directions, e.g. of crystal surfaces, crystal needles or fibers, provide an important signature of the material. This method is also used in the diagnosis of bio-crystals (e.g. gout and pseudo-gout).
Fig. 4-8 Determination of the polarization direction nγ' in a synthetic fiber
(2) Instrumentation
− Eyepiece with graticule
− Tension-free objectives
− Rotary stage (
649H649HFig. 4-7/1)
− Polarizer D (rotatable or fixed), if not using a condenser with an integrated polarizer
− Analyzer module ACR P&C for transmitted light in the reflector turret/slider or analyzer slider D
fixed or with Lambda plate
− Adjustment sample for polarization microscopy (453679-0000-000)
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(3) Adjusting the microscope
• Adjust the microscope as described in chapter
650H650H4.1.1 (3) for transmitted light/bright-field. Make sure
the eye distance is adjusted correctly on the binocular tube (see chapter
651H651H3.5.1).
• Center the rotary stage Pol (
652H652HFig. 4-7/1).
• Swing the polarizer (
653H653HFig. 4-7/3) in the beam path and position it to 0° if you are using a rotatable
polarizer.
• Swing the analyzer module on the reflector turret in (
654H654HFig. 4-7/2) (or slide the analyzer slider into the
intermediate plate). Because of the crossed polarizers the field of view now appears dark.
• Place the adjustment sample Pol on the microscope stage and turn until the sample appears dark.
• Switch off the analyzer and align the graticule along the split cracks of the object.
• Now switch the analyzer back on and remove the sample. The forward direction of polarizer and
analyzer are now parallel to the graticule (Polarizer EW, Analyzer NS).
• Turn the rotary stage Pol with the sample, e.g. a synthetic fiber, so that the sample reaches maximal darkness. The fiber is now parallel to one of the two graticule directions. If the deflection is significant (5° and more) you will need to use a polarization microscope.
Do not change the eyepiece distance on the binocular tube any further in order to avoid shifting the angular position of the graticule to the fiber.
• Now turn the stage by approx. 45° until the longitudinal axis of the fiber is pointing in NE­SW direction (
655H655HFig. 4-9). The sample now shows
the strongest brightness (diagonal position). The sample can have any color in this position.
• Slide in the compensator λ.
Like the sample, the compensator λ is a birefringent object, but it has a defined path difference of 550 nm and a maximum oscillation direction nγ pointing strongly to NE-SW.
When the compensator λ is put in, the sample changes its color depending on its orientation (NE­SW or NW-SE).
The changes in color are based on optical interference. It is necessary to compare the interference colors (phase differences) in both diagonal positions (NE-SW and NW-SE).
Fig. 4-9 Diagram of the color tables
according to Michel-Lévy
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The phase difference results from the interference of the polarization of the sample and the polarization of the compensator λ.
The larger phase difference occurs when the polarization direction of the sample or the absolute or relative largest index of refraction (nγ or nγ') is parallel to the largest polarization direction of the
compensator λ. The sample appears then e.g. in blue-green.
The smallest phase difference occurs if the direction of polarization of the sample with the absolute or relative smallest index of refraction (nα or nα') is perpendicular to the polarization direction of the
compensator λ. The sample then appears e.g. yellow.
(4) Conclusions
The color grey-white which occurs in the bright position as was discussed in the example above, corresponds according to the Michel-Lévy color table (
656H656HFig. 4-9) to a phase difference of 150 nm
The surrounding of the artificial fiber which is not birefringent shows after insurgence of the compensator λ a bright red which corresponds to a phase difference of the compensator of 550 nm (the interference color 1st order for the phase difference of 550 nm corresponds to 1λ).
If the polarization direction of the birefringence sample which is to be investigated is parallel to the largest polarization direction of the compensator λ, i.e. in NE-SW direction, the phase difference of the sample (i.e. grey-white: 150 nm) and the phase difference of the compensator λ (red: 550 nm) add up.
This leads to a change in color of the sample from grey-white to blue-green (resulting phase difference = 700 nm).
If the polarization direction of the birefringence sample which is to be investigated is perpendicular to the largest polarization direction of the compensator λ, i.e. in NW-SE direction, the phase difference of the compensator λ (red: 550 nm) is subtracted from the phase difference of the sample (e.g. grey-white: 150
nm). This leads to a change in color of the sample from grey-white to orange (resulting phase difference = 400 nm).
Color tables according to Michel-Lévy can be ordered under ord. no. 42-312.
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4.1.7 Adjusting the Reflected Light/Bright-Field According to KÖHLER
(1) Application
The reflected light/bright-field microscopy is the easiest and most commonly used optical microscopy method. It is used to examine optically opaque specimens or samples as e.g. cut material or wafers.
For an imaging result which is as true to the object as possible we need not only consider the so called direct bundled beams but just as well the indirect ones, i.e. the beams which diffract and scatter on the sample details. According to ABBE, the image is more true to the object when the fraction of the indirect bundled beams is larger.
The bundled light emerging from the reflected light illuminator is reflected on a color-neutral beam splitter before it passes through the objective which is focused on the specimen surface (so-called condenser function). The objective collects the light reflected on the object and creates, with the tube lens, the microscopic intermediate image. This image can then be examined visually or documented objectively.
(2) Instrumentation
− Axio Scope.A1 with adjusted halogen lamp HAL 100 mounted on the reflected light barrel
− Reflector module bright-field ACR P&C for reflected light in the reflector turret /slider
− Upper stand part HAL 100/HBO 6x HD, DIC with pre-installed aperture and field diaphragm or
upper stand part HAL 100/HBO 6x HD, DIC and 2x diaphragm slider 14x40 mm
(3) Adjusting the reflected light/bright field
− The microscope is in correct operational
mode according to chapter
657H657H3 .
− The microscope is switched on.
• Switch on halogen lamp HAL 100 on the
separate auxiliary power supply unit (
658H658HFig.
4-10/1).
• Adjust light intensity with the control button on the auxiliary power supply unit (
659H659HFig. 4-10/2).
• Put a high-contrast reflected light sample on the microscope stage.
• Swing in objective 10x on the nosepiece (
660H660HFig.
4-11/6).
• Focus the sample with the focusing drive (
661H661HFig.
4-11/5). Try to focus away from the sample in order to avoid any collision between objective and sample.
• Put the knurled button of the aperture diaphragm (
662H662HFig. 4-11/2) to a medium position (approx. half
open and half closed).
Fig. 4-10 External auxiliary power supply
unit for HAL 100
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• Adjust the knurled knob on the field diaphragm (
663H663HFig. 4-11/4) so that the field diaphragm becomes
visible in the field of view (
664H664HFig. 4-11/A).
• Readjust the focus on the edge of the field diaphragm with the focusing drive (
665H665HFig. 4-11/B) and center
the field diaphragm on the edge of the field of view (
666H666HFig. 4-11/C) with the centering screws (667H667HFig.
4-11/3).
• Now open the field diaphragm enough to just make it disappear behind the edge of the field of view (
668H668HFig. 4-11/D).
• For adjusting the aperture diaphragm (image contrast) take out an eyepiece from the tube barrel and look into the barrel or mount an auxiliary microscope instead of the eyepiece.
• Center the aperture diaphragm with the centering screws (
669H669HFig. 4-11/1) and adjust the knurled knob
(
670H670HFig. 4-11/2) to about 2/3 to 4/5 of the diameter of the objective exit pupil (671H671HFig. 4-11/E) for samples
with a medium contrast quality.
In most cases this position of the aperture diaphragm gives the best contrast at almost full resolution which is best compromise for the human eye.
• Replace the eyepiece and adjust the focus of the coaxial rough and fine drive and the image brightness according to the reflected light sample.
Never use the aperture diaphragm to regulate the image brightness! Use the control knob on the auxiliary power supply unit to adjust the light intensity!
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1 Centering screw of the aperture diaphragm 2 Knurled button of the aperture diaphragm 3 Centering screw of the field diaphragm 4 Knurled button of the field diaphragm 5 Focusing drive 6 Nosepiece
Fig. 4-11 Adjusting the microscope in the reflected light/bright-field
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4.1.8 Adjusting the Reflected Light/Dark-Field
(1) Application
The reflected light/dark-field method is applied when samples are examined which do not have areas with different reflectivity (ideal bright-field samples), but which show scratches, cracks, pores (brief: deflections) on the plane surface. All such light scattering details appear bright in the dark-field while the reflective plane areas remain dark.
(2) Instrumentation
− Axio Scope.A1 with adjusted halogen lamp HAL 100 mounted on the reflected light barrel
− Objectives Epiplan-Neofluar, EC Epiplan-Neofluar, Epiplan with the additional label "HD"
− Reflector module dark-field ACR P&C for reflected light in the reflector turret/slider
(3) Adjusting the reflected light/dark-field
• Adjust the microscope as described in chapter 672H672H4.1.7 for reflected light/bright-field. The field diaphragm image should lie just barely outside of the edge of the field of view in order to avoid reflexes.
• Swing reflector module dark-field ACR P&C for reflected light on the reflector turret in the beam path.
• Remove condenser slider 6x20 if mounted.
• Swing in objective position with dark-field objective (HD) on the nosepiece.
• Open aperture diaphragm fully and switch off or remove neutral filters if applicable.
• Put down the sample and adjust the focusing if necessary.
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4.1.9 Adjusting Reflected Light DIC and Reflected Light C-DIC
(1) Application
The reflected light DIC and the reflected light C-DIC methods (DIC = Differential Interference Contrast; C­DIC = Differential Interference Contrast in circular polarized light) are used for the high-contrast imaging of phase objects, i.e. objects which contrary to the amplitude objects, change predominantly the phase of the light.
(2) Instrumentation
− Axio Scope.A1 with adjusted halogen lamp HAL 100 mounted on the reflected light barrel
− Objectives EC Epiplan-Neofluar, Epiplan with the additional label "DIC" or "Pol"
− DIC slider, fitted to the objective engraved on the top surface of the slider with amplification and
aperture or C-DIC slider 6x20 (in connection with the reflector module C DIC P&C, compensator compartment 6x20)
(3) Reflected light DIC, preferably for objectives Epiplan 5x to 100x and LD-Epiplan 10x to 100x
• Adjust the microscope for reflected light/bright-field as described in chapter 673H673H4.1.7. Open the field diaphragm enough so that the edge of the diaphragm lies just barely outside of the edge of the field of view in order to avoid reflexes.
• Swing the reflector module C DIC P&C on the reflector turret/slider in the beam path.
• Swing in objective position with DIC position on the nosepiece.
• Slide the DIC slider in the compartment of the nosepiece (above the objective).
• Put down the sample, focus and turn the mechanical stage so the structure of interest is visible with
maximum contrast.
• The contrast can be optimized with the knurled button on the DIC slider.
(4) Reflected light C-DIC
• Adjust the microscope for reflected light/bright- field.
• Switch reflector module C DIC P&C into the beam path.
• Slide the C-DIC slider (
674H674HFig. 4-12/2) in the
compensator compartment 6x20 (
675H675HFig. 4-12/1).
• Put the sample down, focus and turn the control knob (
676H676HFig. 4-12/4) on the C-DIC slider so
the structure of interest is visible with maximum contrast. No further stage rotation is necessary.
• The contrast can be optimized by adjusting the control knob (
677H677HFig. 4-12/3) on the C-DIC slider.
Fig. 4-12 Compensator compartment 6x20
with C-DIC slider 6x20
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4.1.10 Adjusting Reflected Light TIC
(1) Application
The reflected light TIC method (Micro-interferometry; TIC = Total Interference Contrast in the circular polarized light) is used in imaging and measuring object structures which are on hand in different azimuths.
(2) Instrumentation
− Axio Scope.A1 with adjusted halogen lamp HAL 100 mounted on the reflected light barrel
− Objectives EC Epiplan-Neofluar, Epiplan with additional label "DIC" or "Pol"
− Compensator compartment 6x20
− TIC slider 6x20 with appropriate reflector
module C DIC P&C
(3) Adjusting reflected light TIC
• Put down the sample (e.g. a stepped object) and adjust the microscope for reflected light/bright-field as described in chapter
678H678H4.1.7.
• Swing reflector module C DIC P&C on the reflector turret in the beam path.
• Slide TIC slider 6x20 in the compensator compartment 6x20 (
679H679HFig. 4-12/1). You will see chromatic
interference stripes in the field of view. Using the control knob (
680H680HFig. 4-13/2) of the TIC slider, move the
black interference stripe by sight to the middle of the field of view.
• In order to choose the structure to be measured, turn the knob (
681H681HFig. 4-13/1) on the TIC slider until the
interference stripes are vertical to the direction in which the object is broken down (see
682H682HFig. 4-14). The
interference stripes can be shifted with the aid of the control button (
683H683HFig. 4-13/2).
The step-height d is then determined with the following formula:
a2
b
2
n
d
λ
=
Δ
=
With: d = step-height in nm n = refractive index of the environment, mostly air (n = 1)
Δ = phase difference a = distance between interference stripes b = offset of the interference stripes along the step
λ = wave length of the illumination in nm
Fig. 4-13 TIC slider 6x20
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The values a and b (see
684H684HFig. 4-14) are determined
with the aid of the eyepiece graticule micrometer or with the micrometer eyepiece.
If you work in the white light (without interference filter), you must set λ = 550 nm. When
interference filters are used, it is important to apply the focal point of their wave lengths.
The measured path difference depends on the aperture and increases with the illumination aperture.
The following correction values must be considered depending on the used objective:
Objective Correction factor k
5x/0.15 1.0057
10x/0.25 1.0161
10x/0.30 1.0236
20x/0.4 1.0436
20x/0.50 and 50x/0.75 1.0718
50x/0.60 1.1111
50x/0.75 and100x/0.75 1.2038
50x/0.80 1.2500
50X/0.90 and100x/0.90 1.3929
100X/0.95 1.5241
Table 1: Correction depending on aperture
Example:
a = 11 mm b = 5 mm λ = 550 nm objective 20x/0.50
nm134
mm22
0718.1mm5nm550
a2
kb
d =
⋅⋅
=
⋅⋅λ
=
b
a
Fig. 4-14 Interference stripes
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Attention:
• If the step and its surrounding are made from different materials, the phase jumps characteristic for the material have to be considered. For all non-conducting materials the phase jump is 180°, and for all semi-conductors only slightly different from 180°. Consequently, errors in the step-height determination may be neglected. However, if metals on top of glass are investigated, the results may become erroneous. The phase jumps given in table 2 were calculated for vertical light incidence and compact materials. They can serve as approximate values, since the phase jumps depend on the layer­thickness and the angle of incidence of the light. An accurate determination of the layer-thickness is only possible when the complete object is covered with a homogeneous layer and the path differences are measured.
• If the layers, respectively the steps are transparent, as e.g. in silicon dioxide on silicon, the interference stripes can change their color and consequently the determination of the order of the interference may become problematic. This complication can be avoided if the sample is covered with a homogeneous layer.
Material
Phase jump φ
Copper 140.0°
Gold 142.5°
Silver 151.0°
Bismuth 151.0°
Nickel 157.0°
Iron 157.5°
Zinc 159.0°
Platinum 160.0°
Aluminum 160.0°
Tin 160.5°
Chrome 165.0°
For a thickness measurement, half the difference of the phase jump at the respective interface has to be considered:
22
d
δ
φ
−
Δ
=
Example: extreme case copper on glass
°
=
Φ
140
copper
, °
=
Φ
180
glass
, consequently we
obtain for the additional thickness due to the phase jump
°=
δ
φ
20
2
or
nm30
18
=
λ
Without consideration of the phase jump at the respective interfaces, the thickness value would be too large by 30 nm.
Coal 160.0°
Graphite 165.0°
Silicon 177.0°
Glass 180.0°
Table 2: Calculated phase jumps for
compact material and vertical incidence of light
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4.1.11 Adjusting Reflected Light Polarization – Proof of Bireflexion and Reflexion pleochroism
(1) Application
The reflected light polarization is a contrasting method suited for cut surfaces of mineral ore, coal, ceramics, special metals and alloys. Depending on the orientation of the crystals and the object details, the cut surfaces often react differently when reflected in linearly polarized light.
The illumination light is polarized by the polarizer before passing through the objective onto the specimen surface where it is reflected. Now the beam parts experience phase differences depending on the structure and polarization optical rotations which, when passing through the analyzer, are
represented by different shades of grey. With the aid of a compensator with λ-plate the grey contrast can be converted into a color contrast. Attention! Do not use a compensator 6x20 for this conversion!
A rotatable λ/4 plate in front of the objective (antiflex cap) helps eliminate, even when examining "dark" specimen surfaces, the reflexes which are inevitable when working with objectives with very low scale figures.
(2) Instrumentation
− Axio Scope.A1 with mounted and adjusted halogen lamp HAL 100
− Rotary stage Pol
− Objectives Epiplan-Neofluar Pol, EC Epiplan-Neofluar Pol, Epiplan Pol
− Reflector module DIC P&C or DIC Rot I P&C in the reflector turret/slider
or reflector module Pol P&C plus analyzer slider or analyzer slider plus polarizer slider
(3) Adjusting reflected light polarization
• Adjust the microscope for reflected light/bright-field as described in chapter 685H685H4.1.7.
• If using the objective position with DIC position, remove the DIC slider, if necessary.
• Swing into the beam path the reflector module DIC P&C (
686H686HFig. 4-15/4) on the reflector turret, swing in
the reflector module Pol P&C and slide the analyzer slider into the compartment; or slide the analyzer slider (
687H687HFig. 4-15/1) and the polarizer slider (688H688HFig. 4-15/2) into their compartments, if applicable. When
using the fixed sliders, the polarizer must be orientated in EAST-WEST direction and the analyzer in NORTH-SOUTH direction.
• Put down the sample, adjust the desired magnification, focus, and examine the sample in the
polarization contrast which is now present while turning the rotary stage Pol (
689H689HFig. 4-15/3).
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A sample is bireflective when the sample details show differences in brightness and color which change when the stage rotates.
For samples with low bireflexion we advise using the analyzer with Lambda plate, rotatable.
Pleochroism is present when the color of the sample changes as soon as the stage rotates (overhead polarizer is turned on, analyzer is turned off).
1 Analyzer slider D/A 2 Polarizer slider A 3 Rotary stage Pol 4 Reflector module in the reflector turret/slider
Fig. 4-15 Components for reflected light polarization
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4.1.12 Adjusting Reflected Light Fluorescence
(1) General principle of operation
The reflected light fluorescence method is used to show fluorescent substances in typical fluorescence colors in high contrast. The light originating from a high-performance illuminator in a reflected light fluorescence microscope passes through a heat protection filter onto an excitation filter (bandpass). The filtered short-wave excitation radiation is reflected by a dichroitic beam splitter and is focused on the sample through the objective. The sample absorbs the short-wave radiation before emitting longer-wave fluorescence radiation (Stokes’ Law). This radiation is then captured from the image side by the objective and passes through the dichroitic beam separator. Last, the beams pass through a band-elimination filter (longpass/bandpass) and only the long-wave radiation emitted by the sample passes.
The spectra of the excitation and the band-elimination filters must match very closely. They must be inserted in a reflector module FL P&C together with the according dichroitic beam splitter.
(2) Instrumentation
− Appropriate objectives, e.g. EC Plan-Neofluar or Fluar (UV-excitation)
− Reflector module FL P&C and safety shutter in the upper part (additional shutter function with the
filter slider is optional)
− Lamp HBO 100 or HBO 50, Colibri or HXP 120 for reflected light illumination for reflected illumination
− Fluorescence protection shield
The mercury vapor short arc lamp must be adjusted before starting the reflected light fluorescence method. Re-adjusting the adjustment might become necessary depending on the length of operating time.
(3) Adjusting reflected light fluorescence
Adjusting the reflected light fluorescence is facilitated when starting with a objective of average degree of magnification, e.g. EC Plan-Neofluar 20x/0.50, and a sample of high fluorescence. The use of demonstration samples is also advisable for the start-up.
If necessary, remove the compensator λ, which was used for the transmitted light polarization method, from its compartment before adjusting the reflected light fluorescence.
• Slide the fluorescence protection shield in the compensator compartment above the nosepiece.
• Switch on the objective EC Plan-Neofluar 20x/0.50.
• Turn the condenser turret to the position H transmitted light/bright-field first (or choose phase
contrast if using a Ph objective) and go to the sample detail to be examined.
• Keep the light path in the reflected light illuminator blocked via the elimination position of the filter slider reflected light (
690H690HFig. 4-16/2).
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• Switch on the lamp HBO 100 or HBO 50 (
691H691HFig. 4-16/1) and let it warm up to operational
temperature for about 15 min.
• On the reflector turret/slider (
692H692HFig. 4-16/6),
choose the reflector module FL P&C with the desired fluorescence filter combination (depending on the excitation mode) and switch it on.
• Now unblock the light path in the reflected light illuminator with the filter slider reflected light (
693H693HFig. 4-16/2).
• If necessary, adjust FL attenuator to 100 % transmission in order to facilitate locating fluorescence signals. Reduce the transmission later on in order to preserve the sample.
• Take out an eyepiece from the tube and adjust the aperture diaphragm at sight. Open the compartment for the aperture diaphragm for P&C (
694H694HFig. 4-16/3) enough to see the whole
objective exit pupil.
• Replace the eyepiece in the tube and close the field diaphragm (
695H695HFig. 4-16/4) enough to make it
visible in the field of view.
• Using both centering screws (
696H696HFig. 4-16/5), center the field diaphragm on the edge of the field of view.
• Open the field diaphragm enough to make it just disappear behind the edge of the field of view, or, if you are using a sample which might bleach out, reduce the field diaphragm into the field of view.
• Focus the sample again and optimize the collector position of the HBO 100. Adjust the collector so that the reflector module of the short-wave excitation illuminates the field of view consistently. A correction of the collector position is not necessary in modules with longer-wave excitation.
Fig. 4-16 Components for reflected light
fluorescence
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4.2 Operating and Functional Elements of Optional Components
4.2.1 Lower Stand Part for LED Transmitted Light Illumination
1 LED illumination (mounted on condenser carrier) 2 Cord guide to the connector socket for LED illumination in the stand base 3 Focusing drive – fine adjustment (both sides) 4 Focusing drive – rough adjustment (both sides)
5 Light intensity control (both sides) 6 Power input with fuse compartment 7 Release tractor for vertical stop of focusing drive 8 On/off switch
Fig. 4-17 Lower stand part for LED transmitted light illumination
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4.2.2 Lower Stand Part with HAL 50 Illumination
1 Field diaphragm 2 Filter wheel 6-fold (both sides) 3 Focusing drive – fine adjustment (both sides) 4 Focusing drive – rough adjustment (both sides) 5 Light intensity control (both sides)
6 Power input with fuse compartment 7 Halogen lamp HAL 50 8 Release tractor for vertical stop of focusing drive 9 Compartment for filter slider (both sides) 10 On/off switch
Fig. 4-18 Lower stand part with HAL 50 illumination
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4.2.3 Lower Stand Part for HAL 100 Illumination
1 Field diaphragm 2 Filter wheel 6-fold (both sides) 3 Focusing drive – fine adjustment (both sides) 4 Focusing drive – rough adjustment (both sides) 5 Light intensity control (both sides) 6 Connector for internal power adjustment for auxiliary power supply unit SNT 12 V DC 100 W
7 Connecting nozzle for halogen lamp HAL 100 8 Release tractor for vertical stop of focusing drive 9 Compartment for filter slider (both sides) 10 Gear shift for diffusion disk
Fig. 4-19 Lower stand part for HAL 100 illumination
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4.2.4 Stand Column Vario 380 mm or 560 mm with Gear Box
1 Hand wheel for vertical adjustment of gear box of stand column, 2 Gear box Vario ready for upper stand part 3 Focusing drive – rough adjustment 4 Focusing drive – fine adjustment 5 Release tractor for vertical adjustment (both sides)
Fig. 4-20 Stand column Vario
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4.2.5 Upper Stand Part Transmitted Light
1 Nosepiece 6x bright-field 2 Reflector insert (exchangeable)
4.2.6 Upper Stand Part FL/HBO
1 Field diaphragm (centerable) 2 Port for adjustment aid 3 FL attenuator for fluorescence illumination 4 Compartment for filter slider A 5 Connecting piece for illumination unit 6 Nosepiece 3x bright-field, 3x DIC 7 Reflector insert (exchangeable)
Fig. 4-21 Upper stand part transmitted light
Fig. 4-22 Upper stand part FL/HBO
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4.2.7 Upper Stand Part FL-LED
1 Reflector turret 4-fold (fixed) 2 Control wheel for illumination intensity and on/off switch of LED illumination 3 Pilot lamp LED illumination on/off 4 Nosepiece 3x bright-field, 3x DIC
The upper stand part FL-LED comes equipped with an integrated LED illumination. The LED modules can be exchanged (see chapter
697H697H3.4.9).
The LED illumination is controlled by a wheel which has a push and a turn mode.
− Push the wheel and the LED currently coupled into the beam path is switched on or off.
− The pilot lamp burns when the LED is turned on.
− Turn the wheel to control the light intensity of the currently active LED: up - increase, down - decrease.
− The active LED blinks twice when it reaches its maximum brightness.
− The chosen degree of brightness remains active even after changing to another LED module.
Fig. 4-23 Upper stand part FL-LED
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