Leica DM5000B, DM4000M User Manual

Download

Page 1

Leica DM4000 B Leica DM4000 M Leica DM4500 P Leica DM5000 B

Operating Manual

Bedienungsanleitung

Page 2

Published September 2007 by: Herausgegeben September 2007 von:

Leica Microsystems CMS GmbH Ernst-Leitz-Straße D-35578 Wetzlar (Germany)

Responsible for contents: Verantwortlich für den Inhalt: Dr. Jasna Roeth, Stefan Motyka (Marketing CM, Compound Microscopy, Product Management) (Marketing CM, Compound Microscopy, Produktmanagement) Holger Grasse (Safety Officer according to MPG §30) (Sicherheitsbeauftragter nach MPG §30) In case of questions, please contact the hotline: Bei Fragen wenden Sie sich bitte an die Hotline:

Phone +49(0)64 41-29 2286 Fax +49(0)64 41-2922 55 E-Mail: MQM-Hotline@leica-microsystems.com

Page 3

Leica DM4000 B Leica DM4000 M Leica DM4500 P Leica DM5000 B

Operating Manual

Page 4

Copyrights

All rights to this documentation are held by Leica Microsystems CMS GmbH. Reproduction of text or illustrations (in whole or in part) by print, photocopy, microfilm or other method (including electronic systems) is not allowed without express written permission from Leica Microsystems CMS GmbH.

The term "Windows" may appear in the following text without further identification. It is, however, a registered trademark of Microsoft Corporation. The names of companies and products used herein may be trademarks of their respective owners.

The instructions contained in the following documentation reflect state-of-the-art technology and knowledge standards. We have compiled the texts and illustrations as accurately as possible. Nevertheless, no liability of any kind may be assumed for the accuracy of this manual’s contents. Still, we are always grateful for comments and suggestions regarding potential mistakes within this documentation.

The information in this manual is subject to modification at any time and without notification.

Page 5

Contents

1. Important notes about this manual ....... 7

2. Intended purpose of the microscope .... 8

3. Safety notes................................................ 9

3.1 General safety notes................................. 9

3.2 Electrical safety ......................................... 10

3.3 Disposal....................................................... 11

4. Overview of the instrument .................... 12

5. Unpacking the microscope..................... 17

6. Assembling the microscope ................... 19

6.1 Specimen stage ......................................... 19

6.2 Condenser ................................................... 21

6.3 Tube and eyepieces .................................. 22

6.4 Objectives ................................................... 23

6.5 Light sources for the transmitted

light axis ...................................................... 23

6.6 Light sources for the incident

light axis ...................................................... 25

6.7 Equipping the incident light

turret disk .................................................... 30

6.8 Polarizer and analyzer .............................. 31

6.9 DIC prisms ................................................... 32

6.10 Optional accessories ................................ 33

6.11 Connection to the power supply ............ 34

6.12 Connecting to the CTR5000

electronics box .......................................... 34

7. Startup ......................................................... 35

7.1 Functional principle .................................. 35

7.2 Switching on the unit ................................ 38

7.3 The display

(DM4000 B/4500 B/4000 M/4500 P)......... 39

7.4 The function keys ...................................... 40

7.5 Köhler illumination .................................... 41

7.5.1 Transmitted light ............................. 41

7.5.2 Incident light.................................... 42

7.6. Checking the phase contrast rings ........ 44

7.7 Setting the motorized polarizer

(DM4500 P/DM5000 B) .............................. 45

7.8 Adjusting the light sources...................... 45

8. Operation .................................................... 51

8.1 Switching on the unit ................................ 51

8.2 Stages and object displacement............ 51

8.3 Focusing ...................................................... 53

8.4 Tubes....................................................... 53

8.5 Eyepieces .................................................... 55

8.6 Objectives ................................................... 55

8.7 Magnification changer ............................. 58

8.8 HC P 1x/1.6x tube optics........................... 58

8.9 Light sources .............................................. 59

8.10 Aperture diaphragm and

field diaphragm .......................................... 59

Page 6

Contents

9. Contrast methods for Leica DM4000 B/DM4500 B/

DM4500 P/DM5000 B ................................ 60

9.1 Transmitted light ........................................ 60

9.1.1 Bright field ......................................... 60

9.1.2 Phase contrast.................................. 60

9.1.3 Dark field............................................ 61

9.1.4 Polarization........................................ 61

9.1.4.1 Manual method............................. 61

9.1.4.2 DM4500 P - examinations

in polarized transmitted light...... 62

9.1.4.3 Motorized method ........................ 68

9.1.4.4 Combined methods....................... 68

9.1.5 Differential interference

contrast ............................................ 68

9.1.5.1 DM4500 B/DM4500 P .................... 68

9.1.5.2 DM5000 B........................................ 69

9.2 Fluorescence.............................................. 70

10. Contrast methods for

Leica DM4000 M ........................................ 71

10.1 Incident light .............................................. 71

10.1.1 Bright field ....................................... 71

10.1.2 Dark field.......................................... 71

10.1.3 Polarization...................................... 72

10.1.4 Interference contrast .................... 73

10.2 Transmitted light ........................................ 73

10.2.1 Bright field ....................................... 73

10.2.2 Polarization...................................... 73

11. Troubleshooting......................................... 74

12. Care of the microscope ........................... 77

12.1 Dust cover ................................................... 77

12.2 Cleaning ....................................................... 77

12.3 Handling acids and bases ....................... 78

13. Essential wear and spare parts ............. 79

14. Abbreviations and pictograms ............... 80

15. Index ............................................................ 81

16. EU Declaration of Conformity................. 82

Page 7

1. Important notes about this manual

Caution!

This operating manual is an essential component of the microscope, and must be read carefully before the microscope is assembled and put into operation.

Text symbols, pictograms and their meanings:

(1.2)

→ p.20

This operating manual contains important instructions and information for the operational safety and maintenance of the microscope and accessories. It must therefore be kept safely for future reference.

Numbers in parentheses, such as "(1.2)", correspond to illustrations (in the example, Figure 1, Item 2).

Numbers with pointer arrows (for example → p.20), point to a certain page of this manual.

Caution! Special safety instructions within this manual are indicated with the triangle symbol shown here, and have a gray background.

Caution! The microscope and accessories can be damaged when operated incorrectly.

Explanatory note.

Instructions on disposing of the microscope, accessory components and consumables.

Item not contained in all configurations.

Page 8

2. Intended purpose of the microscope

The DM4000 – DM5000 microscopes to which these operating instructions belong, and which have the identifying letter B, are intended for biological routine and research applications. This includes examining specimens taken from the human body for the purpose of gaining information about physiological or pathological conditions or inborn anomalies, or testing for safety and compatibility for potential recipients, or for monitoring therapeutic measures.

The microscopes that have the identifying letters M or P are intended for materials science, geological or mineralogical examinations.

The manufacturer assumes no liability for damage caused by, or any risks arising from using the microscopes for other purposes than those for which they are intended or not using them within the specifications of Leica Microsystems CMS GmbH. In such cases the declaration of conformity shall cease to be valid.

Caution!

The above-named microscopes comply with the Council Directive 98/79/EEC concerning in vitro diagnostics. They also conform to the Council Directives 73/23/EEC concerning electrical apparatus and 89/336/EEC concerning electromagnetic compatibility for use in an industrial environment.

These (IVD) instruments are not intended for use in the patient environment defined by DIN VDE 0100-710. Nor are they designed to be combined with medical instruments in accordance with EN 60601-1. If a microscope is electrically connected to a medical instrument in accordance with EN 60601-1, the requirements defined in EN 60601-1-1 shall apply.

Page 9

3. Safety notes

3.1 General safety notes

This safety class 1 device was built and tested in accordance with the safety regulations for electrical measuring, control, regulating and laboratory devices in accordance with EN 61010-2-101:2002 EN 61010-1:2001 IEC 1010-1:2001

Caution!

3. Safety notes

Caution!

The devices and accessories described in this operating manual have been tested for safety and potential hazards. The responsible Leica affiliate or the main plant in Wetzlar, Germany, must be consulted whenever the device is altered, modified or used in conjunction with non-Leica components that are outside of the scope of this manual.

In order to maintain this condition and to ensure safe operation, the user must follow the instructions and warnings contained in this operating manual.

Unauthorized alterations to the device or noncompliant use shall void all rights to any warranty claims and void product liability!

Page 10

3. Safety notes

3.2 Electrical safety

General specifications

Leica CTR5000 electronics box (for DM5000 B)

For indoor use only. Supply voltage: Frequency: Power input: Fuses:

90-250 V~ 50-60 Hz max. 290VA T6.3 A

(IEC 60127-2/3) Ambient temperature: Relative humidity: Overvoltage category: Pollution degree:

15-35°C

max. 80% to 30°C

Microscope

For indoor use only. Supply voltage: Frequency:

90-250 V~

50-60 Hz Power input:

DM4000 DM4500 DM5000

max. 180 VA

max. 290VA Fuses:

DM4000 DM4500

DM5000 Ambient temperature: Relative humidity: Overvoltage category: Pollution degree:

T6.3 A (IEC 60127-2/3) T6.3 A (IEC 60127-2/3) See CTR5000 15-35°C max. 80% to 30°C II 2

ebq 100 supply unit*

For indoor use only. Supply voltage: Frequency: Power input: Fuses: Ambient temperature: Relative humidity: Overvoltage category: Pollution degree:

90-250 V~ 50-60 Hz max. 155VA 2xT2A (IEC 127) 10-36°C max. 80% to 30°C II 2

(see enclosed manual)

Caution!

The power plug may only be plugged into an outlet equipped with a grounding contact.

Do not interfere with the grounding function by using an extension cord without a ground wire. Any interruption of the ground wire inside or outside of the device, or release of the ground wire connection, can cause the device to become hazardous. Intentional ground interruption is not permitted!

Caution!

Through connection to the grounding connection, ancillary equipment with its own and/or extra power supply may be brought to the same ground wire potential. For connections without a ground connector, Leica Service must be consulted.

Page 11

Caution!

Never use any fuses as replacements other than those of the types and the current ratings listed here. Bypassing fuse holders is not permitted.

Caution!

The microscope’s electrical accessory components are not protected against water. Water can cause electric shock.

Caution!

Protect the microscope from excessive temperature fluctuations. Such fluctuations can lead to the accumulation of condensation, which can damage the electrical and optical components. Operating temperature: 15-35°C

3. Safety notes

3.3 Disposal

Once the product has reached the end of its service life, please contact Leica Service or Sales about disposal.

Please observe and comply with the national and federal laws and regulations that are equivalent to EU guidelines such as WEEE.

Note!

Like all electronic devices, the microscope, its accessory components and consumables must never be disposed of with general household waste.

Caution!

Before exchanging the fuses or lamps, be absolutely certain to switch off the main power switch and remove the power cable.

Page 12

4. Overview of the instrument

Specification

Contrast methods

Transmitted light axis

Incident light axis

Leica DM4000 B Leica DM5000 B

• Transmitted light: DM4000 B: BF, DF, PH, Pol DM5000 B: and ICT (mot.)

• Incident light: Fluorescent

• Automatic illumination manager (mot. aperture diaphragm and field diaphragm, mot. intensity control)

• Automatic constant-color intensity control

• Motorized shutter

• Integrated into the stand

• Motorized 5x filter turret disk (DM5000 B 8x optional)

• With FIM (fluorescence intensity manager) for reducing the light intensity in 5 increments

• Mechanical booster lens for increasing fluorescence intensity

• Motorized shutter

Leica DM4000 M Leica DM4500 P

• Transmitted light: DM4000 M: BF, DF, PH, ICT,

Pol

DM4500 P: BF, DF, PH, ICT,

Pol (conoscopy)

• Incident light: BF, DF, ICR, Pol, Fluo

• Integrated into the stand

• Motorized 4x filter turret disk

• Automatic illumination manager

• DM4000 M: motorized shutter

Z pinion

Objective turret

• Manual

• Manual, fully encoded

• DM4000 B: 6x/7x with M25 thread

DM5000 B: 7x (M25) DM5000 B: With object prism disk (3 positions)

• Manual, fully encoded

• DM4000 M: 6x with M32 thread DM4500 P: 6x with M25 thread, centerable, encoded

• Receptacle for DIC prisms and Pol compensators (for DM4000 M: optional)

Page 13

4. Overview of the instrument

Specification

X/Y stage

Tube

Condenser

Magnification changer

(optional)

Controls

Leica DM4000 B Leica DM5000 B

• Manual

• Replaceable specimen stage

• Coaxial drive length: 155 mm

Leica DM4000 M Leica DM4500 P

• Manual

• DM4000 M:

Replaceable specimen stage

•

Coaxial drive length: 140 mm

DM4500 P:

• Replaceable Pol stage

• Manual or motorized (DM4500P: manual)

• Optionally with one or two camera outputs

• DM4500 P: conoscopy module (tube optics HC P1x/1.6x with Bertrand lens, encoded)

• Motorized condenser head

• Condenser disk for the light ring, DF-Stop, DIC prisms

• Automatic Köhler illumination

• Optional polarizer (integrated and motorized)

• Manual

• 3x fully encoded

• 1x; 1.25x; 1.6x

• Manual

• 3x fully encoded

• 1x; 1.5x; 2x

• Operating buttons on the stand for all motorized microscope functions

• Additional variable multifunction keys

• Focus wheels

• LCD

• DM5000 B with Leica SmartTouch

Computer interface

Software tools

• USB2.0

• Leica Application Suite (LAS)

for Windows

2000, XP, Vista

• With plug-ins for:

• Microscope and camera configuration

• Microscope and camera control

• Image acquisition

Page 14

4. Overview of the instrument

Specification

Electronics box Leica CTR5000

Leica DM4000 B Leica DM5000 B

Only for the Leica DM5000 B: Separate operating unit with a power supply for 100 W halogen lamps. See → p.10 (Electrical safety)

Leica DM4000 M Leica DM4500 P

Page 15

4. Overview of the instrument

5 6

Fig. 1 Left side of the stand with the advanced AET22 ErgoTube 1 Eyepiece

2 Eyepiece tube 3 Tube 4 Objective turret with objectives 5 Specimen stage with specimen holder 6 Condenser 7 LCD

8 Field diaphragm operating buttons 9 Transmitted light / incident light switch 10 Aperture diaphragm operating buttons 11 Brightness adjustment buttons 12 Focus wheel with coarse and fine adjustment 13 Variable function keys (preset at the factory) 14 Lamp adjustment window

891011

Page 16

4. Overview of the instrument

21 20 19 18 17

Fig. 2 Right side of the stand with the advanced ErgoTube AET22 15 Lamp housing for incident light

16 Lamp housing for transmitted light 17 Transmitted light filter, optional 18 Transmitted light filter, optional 19 Variable function keys (preset at the factory) 20 x/y coaxial drive, adjustable height 21 Handwheel for fine focus 22 Motorized filter block exchanger

Page 17

5. Unpacking the microscope

The microscope is delivered in two packages.

The stand package contains the following com- ponents:

• Stand with integrated incident light axis and objective turret

• Specimen stage with stage bracket

• Power cable and PC connecting cable

• CD with Leica Application Suite (LAS) software package

• Instructions and list of microscope default settings

The system package contains the microscope’s accessories:

• Tube

• Eyepieces

The external ebq 100 supply unit* is delivered in a separate package.

For the Leica DM5000 B microscope: The Leica CTR5000 electronics box is delivered in a separate package.

First, carefully remove all components from the transportation and packaging materials.

Note:

If at all possible, avoid touching the lens surfaces of the objectives. If fingerprints do appear on the glass surfaces, remove them with a soft leather or linen cloth. Even small traces of finger perspiration can damage the surfaces of optical devices in a short time. See the chapter on "Care of the microscope" → structions.

Caution!

p. 77 for additional in-

• Objectives

• Condenser

• Lamp housings with accessories

• Assembly tools

• Additional microscope accessories such as filter cubes, etc. depending on product configuration

Do not connect the microscope or peripherals to an AC power source at this time under any circumstances!

Page 18

5. Unpacking the microscope

Installation location

Work with the microscope should be performed in a dust-free room, which is free of oil vapors and other chemical vapors, as well as extreme humidity. At the workstation, large temperature fluctuations, direct sunlight and vibrations should be avoided. These conditions can distort measurements and micrographic images.

Allowable ambient conditions Temperature 15-35°C Relative humidity maximum 80% up to 30°C

Microscopes in warm and warm-damp climatic zones require special care in order to prevent the build up of fungus. See the chapter on "Care of the microscope" → p. 77 for additional instructions.

Caution!

Electrical components must be assembled at least 10 cm away from the wall and from flammable substances.

Transport

For shipping or transporting the microscope and its accessory components, the original packaging should be used.

As a precaution to prevent damage from vibrations, the following components should be disassembled and packaged separately:

• Unscrew the objectives.

• Remove the condenser.

• Remove the specimen stage.

• Remove the lamp housings.

• Disassemble the burner of 106 z lamp housing.

• Remove all moving or loose parts.

Page 19

6. Assembling the microscope

6. Assembly

The microscope components are logically assembled in this order:

• Specimen stage

• Condenser with condenser head

• Tube

• Eyepieces

• Objectives

• Lamp housings with light sources

• Equipment for the incident light turret disk*

Only a few commonly used screwdrivers and keys are necessary for assembly; these are included in the delivery package.

When using intermediate systems and optical accessories, the sequence may vary. In this case, read chapter, "6.10 Optional accessories" →

p.33

6.1 Specimen stage

Caution:

Never install objectives before assembling the stage.

• Place the specimen holder on the stage and fasten it with the two screws (3.1).

• Using the condenser height adjuster (3.2), turn the condenser holder completely upwards, i.e. as close to the stage as possible.

• Loosen the stage clamp (3.3) slightly.

Fig. 3 Mechanical object stage 1 Locking screws for specimen holder 2 Condenser height adjuster 3 Stage clamp

Page 20

6. Assembly

• From above, set the stage clamp onto the dovetail guide (4.2) and push the stage downwards until the upper end of the dovetail guide is tightly fastened to the upper end of the stage clamp.

• Firmly tighten the stage clamp (4.1).

Note:

For thicker specimens (Leica DM4000 M) the stage can be set to a correspondingly lower level.

Fig. 4 Assembling the stage 1 Stage clamp 2 Dovetail guide

• Only for DM4500 P:

Pol attachable mechanical stage*

Adjust the attachable mechanical stage so that the fastening screw is visible below the holes (4a.1). Set the attachable mechanical stage in the guide holes on the rotating stage and tighten the fastening screw using the hexagonal key.

Attachable mechanical stage*

The attachable mechanical stage can be installed on the left, on the right or on the front (not pictured). The two clamping screws fasten it into place.

Fig. 4a Pol rotating stage* and Pol 3 attachable mechanical stage*

1 Holes for the fastening screw. 2 Lever for the holder for glass slides of various formats,

which can be turned inward and outward

3 Storage for the centering key 4 Locking button pair 5 45° click stop 6 Clamping system for the stage rotation

Page 21

6.2 Condenser

6. Assembly

• Screw the condenser head into the condenser.

• Using the condenser height adjuster (5.4), turn the condenser holder (5.1) downward as far as it will go.

• Unscrew the clamping screw for the condenser (5.3) far enough so that the condenser can be inserted from the front.

• From the front, insert the condenser into the condenser holder as far as it will go. On the underside of the condenser, there is an orientation pin (6.1) that must be locked into place in the guiding notch (7.1).

• Tighten the condenser’s clamping screw (5.3) until the condenser locks into place.

• Connect the condenser over the connection (8.1) with the stand.

Note:

The condenser must be centered before using the microscope.

Köhler illumination p. 41.

→

Fig. 6

Underside of condenser

1 Orientation pin

Fig. 7 Condenser holder 1 Guiding groove

Fig. 5 Condenser holder 1 Condenser holder 2 Condenser centering 3 Clamping screw for the condenser 4 Condenser height adjuster

23 4

Fig. 8 Condenser connection 1 Condenser cable socket

Page 22

6. Assembly

6.3 Tube and eyepieces

The tube is mounted on the stand either directly or with the use of intermediate modules. The side clamping screw fastens it into place (9b.1).

For the MBDT motorized tube only:

• Remove the transport anchor (9a.1) on the underside of the tube.

• Partially unscrew the clamping screw (9b.1).

• Insert the tube into the circular receptacle (dovetail ring).

• Retighten the clamping screw (9b.1).

For the MBDT motorized tube only:

• Connect the tube to the stand with the connector bushing (10.1).

• The eyepieces are inserted into the eyepiece tubes on the tube.

For the BDTP tube only:

• The Pol eyepieces are inserted into the eyepiece tubes (using the locking groove).

Fig. 9b Fastening the tube 1 Clamping screw

Fig. 9a Underside of the tube 1 Transport anchor

Fig. 10 Motorized tube connection 1 Connector socket

Page 23

6. Assembly

6.4 Objectives

The receptacles on the objective turrets are numbered (Fig. 11). Based on your equipment, the individual objectives have already been assigned to specific positions at the factory. For details on the exact positions of the objectives, please refer to the enclosed identification sheet.

Caution:

Close vacant threads in the turret with dust protection caps!

Fig. 11

Objective turret with engraved objective receptacles

6.5 Light sources for the transmitted light axis

Caution!

Ensure that the lamp housing has been disconnected from the power supply. Unplug the power plug and the power supply during assembly.

Caution!

Light sources pose a potential irradiation risk (glare, UV-radiation, IR-radiation). Therefore, lamps have to be operated in closed housings.

Lamp housing 107/2

This lamp housing is used with a 12 V 100 W halogen lamp, which is already mounted. In case the lamp has to be removed:

• Remove the fastener screw on the housing (Fig. 12).

• Remove the housing by pulling it upwards.

• Remove the lamp.

Page 24

6. Assembly

• Insert the new 12 V 100 W lamp (13.1) with the dust cover straight into the socket until it stops. Be sure that the lamp is inserted straight.

• Remove the lamp’s dust cover.

Caution!

Do not remove the lamp’s dust cover until after you have installed the lamp. Avoid fingerprints on the lamp.

• Replace the housing and fasten it in place using the fastening screw.

Fig. 12

Lamp housing 107/2 Releasing the fastening screw

• Place the lamp housing in the transmitted light lamp housing receptacle (14.2) and fasten it with the clamping screw on the side.

• Connect the lamp housing to the power supply for transmitted light (symbol:

Fig. 14 Rear view of the stand 1 Incident light lamp housing receptacle 2 Transmitted light lamp housing receptacle 3 12 V 100 W connection for transmitted light (symbol: ) 4 12 V 100 W connection for incident light (symbol: )

) (14.3).

Fig. 13

Lamp housing 107/2 opened 1 Mount with

halogen bulb

2 Collector

Page 25

6. Assembly

6.6 Light sources for the incident light axis

Caution!

Light sources pose a potential irradiation risk (glare, UV-radiation, IR-radiation). Therefore, lamps have to be operated in closed housings.

Ensure that the lamp housing has been disconnected from the power supply. Unplug the power plug and the power supply during assembly.

During assembly work on xenon burners, always wear the protective gloves and face protection supplied (Fig. 15) (risk of explosion).

Never touch the glass parts of the burner with bare hands. Never look directly into the beam path (blinding hazard).

Lamp housing 106/106 z

This lamp housing is suitable for use with a 12 V 100 W halogen lamp or a variety of gas discharge lamps.

Caution!

Make sure to follow the instructions and safety notes of the lamp supplier. Before changing lamps allow it to cool down for at least 30 min.!

Fig. 16 106/106 z lamp housing (on the side, open) 1 Cover raised 2 Collector 3 12 V 100 W lamp or

gas discharge lamp in mount

4 Reflector (mirror) 5, 6, 7 Adjusting screw for x-y reflector 8 Fastening screw for the lamp mount 9 Socket for contact plug

Fig. 15

Protective gloves and mask

898

Page 26

6. Assembly

Inserting the 12 V 100W halogen bulb into the 106/106 z lamp housing

• Unscrew the fastening screws of the cover and flip the cover up (16.1).

• Unscrew the fastening screws of the lamp mount (16.8) and pull out the mount (Fig. 17).

• Insert the lamp with the dust cover straight into the socket until it stops.

Caution!

Do not remove the lamp’s dust cover until after you have installed the lamp. Avoid fingerprints on the lamp.

• Remove the dust cover.

Fig. 17 Lamp mount with 12 V 100 W halogen bulb

• Insert the lamp mount, with the burner installed, into the lamp housing and tighten it with the screws (16.8).

• Close the lamp housing and retighten the fastening screws.

• Place the lamp housing in the incident light lamp housing receptacle (18.1) and fasten it with the clamping screw on the side.

• Connect the lamp housing to the power supply for incident light (symbol:

Fig. 18 Rear view of stand 1 Incident light lamp housing receptacle 2 Transmitted light lamp housing receptacle 3 12 V 100 W connection for transmitted light (symbol: ) 4 12 V 100 W connection for incident light (symbol: )

) (18.4).

Page 27

6. Assembly

Inserting gas discharge lamps (Hg and Xe) into the 106/106z lamp housing

Hg and Xe lamps are powered by separate supply units. Please also read the separate instruction manual provided with these supply units.

The following gas discharge lamps may be used and require different power supplies and lamp mounts (Fig. 19):

Type Typical bulb life*

50 W high-pressure mercury burner (alternating current) 100 hrs. 100 W high-pressure mercury burner (direct current) 200 hrs. 100 W high-pressure mercury burner (direct current, type 103 W/2) 300 hrs. 75 W high-pressure xenon burner (direct current) 400 hrs.

* Please observe the data sheets of the lamp manufacturer.

Page 28

6. Assembly

• To open the 106 z lamp housing, unscrew the fastening screws on the cover.

Caution!

• Remove the transport anchor (red plastic rod in place of the burner) in the lamp mount. To do so, remove the lower clamp (19.1). Pull up the cooling element (19.3) and turn it to the side. Detach the lower clamp system (19.2) and remove the transport anchor.

• Install the burner in reverse order.

Fig. 19 a-c Lamp mounts for gas discharge lamps 1 Upper clamping system, 2 Lower clamping system, 3 Cooling element 4 Melt nipple for the Hg 50 arc lamp, 5 Dust cover for the Xe 75 arc lamp

Hg 50

Hg 50 burner:

After installation, the labeling must be right. If a glass melt nipple is present (19a.4), position it by turning the burner so that the nipple does not impede the beam path later, but instead is positioned

Xe 75 burner:

Remove the burner’s dust cover (19b.5) after you have installed the burner.

Xe 75

up-

sideways.

Hg 100

Page 29

• Insert the lamp mount, with the burner installed, into the lamp housing and tighten it with the screws (20.8).

6. Assembly

• Close the lamp housing and retighten the fastening screws.

• Place the lamp housing in the incident light lamp housing receptacle (21.1) and fasten it with the clamping screw on the side.

• Connect the lamp housing to the external power supply (22.1).

Fig. 21 Rear view of the stand 1 Incident light lamp housing receptacle 2 Transmitted light lamp housing receptacle 3 12 V 100 W connection for transmitted light (symbol: ) 4 12 V 100 W connection for incident light (symbol: )

Fig. 20 106/106 z lamp housing (on the side, open) 1 Cover raised 2 Collector 3 12 V 100 W lamp or

gas discharge lamp in mount

4 Reflector (mirror) 5, 6, 7 Adjusting screw for x-y reflector 8 Fastening screw for lamp mount 9 Socket for contact plug

898

Fig. 22 Rear panel of the ebq 100 supply unit 1 Lamp connection

Page 30

6. Assembly

6.7 Equipping the incident light turret disk

The positions in the turret disk are numbered. Depending on how they are equipped, the individual filter and/or reflector cubes are set in pre-assigned positions at the factory. For details, check the identification sheet included with your order.

Insert the filter and reflector cubes in the following manner:

• Never fit the incident light turret disk while the microscope is in operation.

• Remove the face plate from the upper part of the microscope (Fig. 25). Press the locking pin (25.2) to turn the turret disk. When the locking pin is released, the turret disk locks into place again.

• With the holder facing you squarely, insert the filter cube or reflector cube into the holder as described in the identification sheet provided. To do so, place the filter or reflector cube on the right side and press it toward the left into the mounting (Fig. 26).

Fig. 23 Filter cube,

front side

Fig. 25 Removing the front panel 1 Filter receptacle 2 Locking pin 3 Front panel

Fig. 24 Filter cube,

back side

1 2 3

• Press the locking pin (25.2) and turn the filter turret to the next click stop.

• Make sure that the turret engages (the locking pin springs forward) and insert the next filter and/or reflector cube as described above.

• When all filters and reflector cubes have been inserted, close the front cover plate again.

Fig. 26 Inserting the filter or reflector cubes 1 Mounting

Page 31

6. Assembly

6.8 Polarizer and analyzer

Transmitted light polarizer: ICT/P

• Using the left clamping screw, fasten the ICT/P transmitted light polarizer to the underside of the condenser holder (Fig. 27).

• Make sure that the red index point on the front of the polarizer is at 0.

• If necessary, insert the compensators (λ-, λ/4 plates or elliptical compensators) into the polarizer’s receptacle (Fig. 28) or into the compensation slot.

Incident light polarizers:

R/P polarizer, rotating polarizer,

L/ICR polarizer, R/ICR polarizer

Fig. 27 Assembly of the ICT/P transmitted light polarizer 1 Clamping screw

Fig. 28 Inserting the compensators

• Remove the plug cap on the right side of the incident light axis (Fig. 29).

• Insert the polarizer into the receptacle until it locks into place.

Caution:

Push the polarizer into the front receptacle only.

Motorized polarizer

• A motorized polarizer is already installed and ready for operation in the DIC condenser.

Fig. 29 Inserting the polarizer 1 The plug cap is replaced with the polarizer.

Page 32

6. Assembly

Transmitted light and incident light analyzer

• Remove the plug cap on the left side of the stand.

• Insert the analyzer into the receptacle until it latches in place (Fig. 30).

Motorized analyzer

• Insert the analyzer cube into the corresponding position on the filter turret as described in section "6.7 Equipping the incident light turret disk" → p. 30. For details on the proper position for the analyzer cube, please refer to the identification sheet provided.

6.9 DIC prisms

• Insert the objective prism slide into the tube slot (Fig. 31.1). The code letter must match the code letter on the objective.

• In the Leica DM5000 B microscope, the DIC prisms are already installed in the DIC disk above the objective turret (Fig. 68).

Fig. 30 Inserting the analyzer 1 The plug cap is replaced with the analyzer.

Fig. 31 Inserting the objective prism slide 1 Objective prism slide

Page 33

6. Assembly

6.10 Optional accessories

ErgoModule

For raising the eye level of the tube opening, the ErgoModule may be used. It is fastened in place with the side clamping screw.

Mirror housing

• Place the mirror housing directly onto the lamp housing receptacle on the back of the stand and attach it using the side clamping screw.

• Place the lamp housing(s) on the mirror housing and fasten it using the accompanying lateral clamping screw.

Booster lens / excitation manager

• Insert the filter slide into the front receptacle on the right side of the stand (Fig. 32.1, 33.1).

Fig. 32 1 Inserting the booster lens

Page 34

6. Assembly

6.11 Connection to the power supply

After completing the assembly work, connect the stand to the power supply using the power cable (Fig. 34.2).

Fig. 34 Rear side of the Leica DM4000 B/M stand 1 Power switch 2 Power supply

Fig. 35 CTR5000 electronics box connector panel 1 Microscope connection 2 Power supply

6.12 Connection to the CTR5000 electronics box

For the Leica DM5000 B only:

• Connect terminals (35.1) and (36.1) to the 25-pin microscope cable.

• Connect the electronics box to the power supply using the power cable (35.2).

Fig. 36 Rear side of the Leica DM5000 B stand 1 Connecting to the CTR5000 electronics box

Page 35

7. Startup

7.1 Functional principle

The microscope's most important functions may be easily accessed using function keys.

• The microscope may be switched between various contrast methods by pressing a button.

• The microscope recognizes the selected objective and associated contrast method. Therefore, the values for intensity (INT), aperture diaphragm (AP) and field diaphragm (FD) are always set correctly.

• The INT, AP and FD values can be adjusted individually. Manual adjustments overwrite the previous settings. The current setting is saved.

• The INT, AP and FD values are always based on the currently activated light axis (transmitted light or incident light).

• In addition to the preset function keys for INT, AP and FD, there are also variable function keys.

Variable function keys:

• At the time of delivery, these function keys are assigned functions suitable to the configuration of your microscope.

• However, the functions can be reprogrammed and/or adapted to your specific requirements.

Note: (reset function)

The microscope can be reset to its factory default programming:

When the microscope is switched off, press all 3 variable function keys on the left stand section.

•

• Switch on the power for the stand.

• Hold the buttons until the initialization is complete.

• The standard indicator appears in the display.

• Switch the instrument off and back on. The settings are now saved.

Page 36

7. Startup

Possible assignments for the function keys

For Leica DM4000 B/DM5000 B:

Function button Function

BF Bright field transmitted light PH Phase contrast transmitted light ICT Interference contrast, transmitted light DF Dark field transmitted light POL Polarization transmitted light CHANGE TL

INT ↑ Increases the brightness (transmitted light) INT ↓ Reduces the brightness (transmitted light) AP ↑ Opens the aperture diaphragm (transmitted light) AP ↓ Closes the aperture diaphragm (transmitted light) FD ↑ Opens the field diaphragm (transmitted light) FD ↓ Closes the field diaphragm (transmitted light)

SHUTTER TL Opens/closes the transmitted light shutter

FLUO Fluorescence (last filter cube) CUBE 1 Chooses the fluorescence cube on position 1 CHANGE CUBE CHANGE CUBE SHUTTER FLUO Opens/closes the fluorescence shutter

INT FLUO ↑ Increases the brightness (fluorescence) INT FLUO ↓ Reduces the brightness (fluorescence) FD FLUO ↑ Opens the field diaphragm (fluorescence) FD FLUO ↓ Closes the field diaphragm (fluorescence)

COMBI

CHANGE COMBI

Switches through all transmitted light contrast methods

Switches through the fluorescence cube clockwise (1 → 4) Switches through the fluorescence cube counterclockwise (4 → 1)

Combination method

(PH fluorescence or ICT fluorescence)

Switches through all combination methods

CHANGE TUBE Switches through various types of beamsplitting 100% VIS 100% documentation port 50:50 50% documentation port / 50% camera 100% CAMERA 100% camera

Page 37

For Leica DM4000 M/DM4500 P:

Function button Function

BF Bright field incident light ICR Interference contrast, incident light DF Dark field incident light POL Polarization incident light CHANGE RL

INT ↑ Increases the brightness (incident light) INT ↓ Reduces the brightness (incident light) AP ↑ Opens the aperture diaphragm (incident light) AP ↓ Closes the aperture diaphragm (incident light) FD ↑ Opens the field diaphragm (incident light) FD ↓ Closes the field diaphragm (incident light)

SHUTTER RL DM4000 M only: Opens/closes the incident light shutter

FLUO Fluorescence (last filter cube) CUBE 1 Chooses the fluorescence cube on position 1 CHANGE FLUO Switches through all filter cubes CONOS DM4500 P only: The

Switches through all incident light contrast methods

Bertrand lens is in the beam path

(conoscopic beam path)

7. Startup

FOCUS FINDER Finds the smallest incident light field diaphragm

and toggles back to the original field diaphragm.

BF TL Bright field transmitted light BF-POL DM4500 P only: Polarization transmitted light (conoscopy) INT ↑ Increases the brightness (transmitted light) INT ↓ Reduces the brightness (transmitted light) AP ↑ Opens the aperture diaphragm (transmitted light) AP ↓ Closes the aperture diaphragm (transmitted light) FD ↑ Opens the field diaphragm (transmitted light) FD ↓ Closes the field diaphragm (transmitted light)

COMBI

CHANGE TUBE Switches through various types of beamsplitting 100% VIS 100% documentation port 50:50 50% documentation port / 50% camera 100% CAMERA 100% camera

Combination methods (BF and BF TL)

Page 38

7. Startup

7.2 Switching on the unit

• First, swivel the objective with the lowest magnification into position.

• Switch on the microscope or CTR5000. All motorized microscope components will then run through an initialization phase.

After the initialization is complete, the display on the stand (Fig. 37) shows the current microscope setting

Caution!

After turning on the gas discharge lamp, the burner must be immediately adjusted. Therefore, do not turn on the power supply unit yet. First, work in transmitted light in order to familiarize yourself with the microscope’s controls.

Components such as diaphragms, condensers, light and phase rings have been pre-centered at the factory. It may be necessary to correct the centering after the microscope has been transported and assembled. Before proceeding with the necessary steps, first familiarize yourself with the stand’s display and control panel.

Note:

The Leica DM4500 P microscope is equipped at the factory with an encoded conoscopy module.

• Moving the Bertrand lens activates the conoscopic or orthoscopic beam path. The status is indicated on the display with the methods "Conos" or "TL Pol".

Fig. 37 Display after initialization

Page 39

7. Startup

7.3 The display (Leica DM4000 B/ DM4000 M/DM4500 P)

The screen displays the microscope’s current settings. The content of the display depends on the features of the individual microscope. In the first column, corresponding symbols indicate the type of information: contrast method, magnification, light intensity, diaphragms, light splitting for photo tubes. Please see the abbreviation index for a list of abbreviations used, →

Contrast method

In the first row, you find an indication of the active light axis (incident light or transmitted light) of the current contrast method and, if used, the current filter cube.

The shutter status display for the transmitted light or incident light shutter:

Transmitted light shutter is open

↑

Transmitted light shutter closed

p. 36f.



The current brightness setting is graphically depicted by a beam. In addition, the light intensity is specified in 20 (coarse) or 255 (fine) increments →



The values for the field diaphragm (FD) and the aperture diaphragm (AP) are indicated numerically. The field diaphragm in the incident light may be either round or rectangular. Accordingly, the FD designation is set in parentheses or in brackets: (FD) or [FD].

When using a digital camera, angular field diaphragms are recommended.

Light intensity

p. 59.

Diaphragm

Note:

Light distribution

Incident light shutter is open

↓

(for fluorescence only)

↓

Incident light shutter is closed (for fluorescence only)

Magnification

The current objective magnification (OBJ), sometimes followed by the re-magnification of the magnification changer*, appears along with the total magnification:

Σ = Objective x re-magnification x eyepiece

If a motorized tube is present, the light division between ocular (Eye) and photo port (Docu) is indicated in %.

Note:

The display may flash after the initialization phase or even during microscopy. This always occurs when the contrast method selected can not be performed with the microscopic settings. For example, an objective may be rotated in that is not suited to the contrast method chosen. Then check your settings.

Page 40

7. Startup

7.4 The function keys

There is a row of function keys both on the right and left side of the stand. These can be broken down into fixed and variable buttons. The variable function keys have different functions depending on the features of the individual microscope.

Fixed function keys on the left side of the stand

The TL/IL button (38.1) toggles between the incident light and transmitted light axis. The contrast method last used with a given axis is restored when switching. The INT buttons (38.3) adjust the light intensity. The adjustment can be made in coarse or fine steps. Pressing both INT buttons at the same time toggles between coarse and fine adjustment. The display indicator changes accordingly → p. 59.

Fig. 38 Fixed function keys (left side of stand) 1 Toggling between transmitted light / incident light 2 Aperture diaphragm 3 Light intensity 4 Field diaphragm

The AP buttons (38.4) for the aperture diaphragm and FD (38.2) for the field diaphragm open and close their respective diaphragms.

Note:

Changes to the light intensity as well as aperture and field diaphragm settings are stored for the individual objectives and contrast methods.

Variable function buttons

The variable function buttons are assigned functions at the factory that are appropriate to the features of your microscope. They are labeled accordingly. For details on button assignments, please refer to the included identification sheet. For information on the abbreviations used, please refer to the list → p. 36f.

Note:

The Leica Application Suite (LAS) module "Configuration" is required for changing the key assignments.

Page 41

7. Startup

7.5 Köhler illumination

7.5.1 Transmitted light

Suitable aperture and field diaphragm values have been preset for each objective. The condenser is also already centered at the factory. However, depending on how the condenser is disassembled and reassembled, it may be necessary to re-center the condenser in some cases. Therefore, check the condenser centering.

The following procedure is provided for the transmitted light-bright field illumination.

• Select an objective with moderate magnifica-

tion (10x-20x).

• Push the TL/IL button (38.1) as needed to acti-

vate the transmitted light axis. "TL" appears in the first line of the display.

Note on DM4500 P: Ensure that the Bertrand lens is swiveled outwards.

• Press the BF button to activate the bright field

contrast method (one of the variable function keys behind the focus wheel). "TL BF" appears in the first line of the display.

• Insert the specimen in the stage’s specimen holder (39.3).

• Focus on the specimen. The focus wheel on the left side of the stand allows focus adjustment in course and fine increments. On the right side of the stand, there is also a focus wheel for fine focusing.

• Adjust the light intensity with the INT buttons (38.3).

• Close the field diaphragm with the FD function key (38.2) until the edge of the diaphragm appears in the specimen plane.

Fig. 39 Stage with specimen holder 1 Object motion (X direction) 2 Object motion (Y direction) 3 Specimen holder 4 Condenser height adjuster

Fig. 40 Condenser centering 1 Centering screws

Page 42

7. Startup

• Using the condenser height adjuster (39.4), adjust the condenser until the edge of the field diaphragm appears in sharp relief.

• If the image does not appear in the middle of the field of view (41c), the condenser must be moved into the middle of the field of view with the help of the two centering screws (40.1).

• Open the field diaphragm just enough for it to disappear from the field of view (41d).

Caution:

The condenser height adjustment depends on the thickness of the specimen. It may need to be adjusted for each specimen.

Fig. 41 Köhler illumination a Field diaphragm not focused, not centered b Field diaphragm focused, but not centered c Field diaphragm focused and centered

Diameter is too small, however

d Field diameter (light) = Field diameter (view)

(Köhler illumination)

7.5.2 Incident light

Suitable aperture and field diaphragm values have been preset for each objective. The incident light module has also been centered at the factory.

However, it may be necessary to readjust the incident light module in some cases after transporting and setting up the stand. Therefore, check the aperture and field diaphragm centering. The following procedure is provided for the incident light-bright field illumination.

• Select an objective with moderate magnification (10x-20x).

• Activate the incident-light axis with the TL/IL button (38.1). "IL" appears in the first line of the display.

• Select the bright field contrast method by pressing the IL-BF button (DM4000 M) or select fluorescence by pressing the FLUO button (DM4000 B, DM5000 B). These functions can be assigned to the variable function keys on the stand. IL BF / FLUO appears in the first line of the display.

• Insert the specimen in the stage’s specimen holder (39.3).

• Focus on the specimen with the focus wheels.

• Adjust the light intensity using the INT buttons (38.3).

Page 43

7. Startup

Adjusting the field diaphragm

• Close the field diaphragm with the FD button

(38.4) or manually until the edge of the diaphragm (round or rectangular) appears in the field of view.

• If the border of the field diaphragm does not appear in the middle of the field of view, the field diaphragm must be moved into the middle of the field of view using the two centering screws (42a.1) on the right side of the stand.

• Use the function buttons FD (38.4) to open the field diaphragm to the point that they just disappear from the field of view.

• We recommend the use of a rectangular field diaphragm when using a digital camera. Match the size of the diaphragm to the chip size of the camera.

Adjusting the aperture diaphragm (for DM4000 M and DM4500 P only)

• Remove one eyepiece (e.g. right).

• Close the aperture diaphragm with the AP

function key (38.2) until the edge of the diaphragm appears in the exit pupil of the objective (aperture diaphragm plane).

• If the image is not in the middle of the field of view of the exit pupil, move the position of the aperture diaphragm to the center of the exit pupil using the two centering screws (42b.2) located on the left side of the stand.

• Open the aperture diaphragm to cover 2/3 of the field of view.

Fig. 42a Adjusting the field diaphragm in the incident light axis 1 Adjusting screws for moving the field diaphragm

Fig. 42b Adjusting the aperture diaphragm in the incident light axis 1 Adjusting screws for moving the aperture diaphragm

Page 44

7. Startup

7.6 Checking the phase contrast rings

If your microscope is equipped for the use of phase contrast, the light rings that fit the objectives are built into the condenser. The light rings are already centered in the factory. However, the centering should be rechecked.

Note:

Every objective is assigned its own light ring in the condenser disk. Therefore, a check must be performed for each objective. When rotating in an objective that is suitable for phase contrast, the corresponding light ring is set automatically.

• Press the BF (bright field) button (one of the

variable function keys, to the left behind the focus wheels).

• In the place of an eyepiece, insert the focusing telescope (Fig. 43) into the observation tube.

• Rotate the phase contrast objective with the lowest magnification into place.

• Focus on the specimen with the focus wheel .

• Focus the ring structure (44a.a) by slightly loosening the clamping ring (43.2) and moving the eyelens (43.1).

• Retighten the clamping ring.

• Press the PH (Phase Contrast) button. The ring diaphragm (light ring) in the condenser is turned inward.

• If the light ring and the phase ring are not shown as arranged in Fig. 44a.c, the light ring must be centered.

Fig. 43 Focusing telescope 1 Adjustable eyelens 2 Clamping ring for fixing the focus position

Fig. 44a Phase contrast centering procedure PH=phase contrast ring, LR=light ring

a Condenser in bright field (BF) position b Condenser in phase contrast (PH) position

Light ring (LR) not centered

c Light ring and phase ring centered

ab c

Page 45

7. Startup

• Insert the centering key through the corresponding openings (44b.1) on both sides of the condenser.

• Turn the centering key until the dark ring (phase ring in the objective) is congruent with the slightly narrower bright ring (light ring in condenser) (44a.c).

Note:

When changing the objective, the centering key must no longer be in the corresponding openings.

• Repeat the process for all other phase contrast objectives.

• Always remove the centering keys after the centering procedure.

7.7 Setting the motorized polarizers (DM4500 P/DM5000 B)

• Select the POL method (one of the variable

function keys on the stand or on the LeicaScreen).

• Insert the centering key into the correspond-

ing openings (44b.2) on the condenser.

• Set the optimum extinction (max. darkness).

7.8 Adjusting the light sources

Transmitted-light axis (TL) with lamp housing 107/2

The lamp housing 107/2 with a 12 V 100 W halogen lamp is fixed. Centering the lamp is not required.

Fig. 44b Light ring centering 1 Centering key in the phase contrast centering hole 2 Polarization contrast centering hole

Fig. 45 Reflector cube for lamp adjustment

Page 46

7. Startup

Incident-light axis (IL) with lamp housing 106 z

•

When a supply unit is used, it is turned on first.

• Activate the incident-light axis with the TL/IL function button. FLUO (Leica DM4000 B/ DM5000 B) or IL (Leica DM4000 M/DM4500 P) appears in the display.

• Insert the lamp adjustment reflector (Fig. 45) in the filter turret in place of a filter cube. Make sure to switch off the instrument first. (See →

p. 30). Make a note of the designation of the replaced filter cube.

Note:

To avoid faulty adjustments, it is a good idea to remove the filter cube located at the left of the reflector cube as well.

• Turn the reflector into the beam path. The reflector has reached the correct position when the name of the exchanged filter cube is shown in the upper right of the display.

In the lamp housing 106z, the direct image of the filament (in halogen lamps) or the arc (in gas discharge lamps) and its reflection are focused separately and adjusted in relation to one another.

On the left side of the microscope, there is an adjustment window (1.14, p. 15) for mapping the light source.

Adjust the lamp as follows while observing the light source in the adjustment window.

Fig. 46 106 z lamp housing 1 Lamp height adjustment

2.4 Mirror image height and side adjustment 3 Focusing the reflector 5 Lamp side adjustment 6 Collector (focusing of the lamp image)

Caution!

Never look directly into the beam path! Beware of the glare hazard when switching to reflector BF or Smith!

Caution!

Light sources pose a potential irradiation risk (glare, UV-radiation, IR-radiation).

516

Page 47

7. Startup

Centering the 12 V 100 W halogen bulb

• In the adjustment window, you see the direct filament image and the mirror image, which as a rule are shifted together.

• Focus the direct filament image with the collector (46.6).

• Use the adjusting buttons on the rear side of the lamp housing (46.2, 46.4) to rotate the mirror image of the lamp filament to the side or completely out of the beam path. The lamp filament’s focused image remains visible (Fig. 47).

• Adjust the direct filament image using the adjusting knobs (46.1) and (46.5) so that the centering surface is halfway covered (Fig. 48).

• Then rotate the mirror image of the lamp filament using the adjusting knobs (46.2) and (46.4), and focus it using the reflector (46.3).

• Align the mirror image symmetrically to the filament image (Fig. 49). To do so, again use the adjusting knobs (46.2) and (46.4).

Fig. 47 Direct lamp filament image focused,

but not centered (in reality, the image is less focused)

Fig. 48 Direct lamp filament image in target position

(in reality, the image is less focused)

• Defocus the image with the collector head (46.6) until the filament image and mirror image are no longer recognizable and the image is uniformly illuminated.

• Replace the lamp adjustment reflector with the original filter cube.

Note:

Make sure to switch off the instrument.

Fig. 49 Direct lamp filament image and mirror image in

target position (in reality, the image is less focused)

Page 48

7. Startup

Centering the Hg 50 W Mercury Lamp

• The adjustment window shows the direct image of the arc and its mirror image. These are generally not in alignment with one another.

• Focus the direct image with the collector (46.6).

• Use the adjusting buttons on the rear side of the lamp housing (46.2, 46.4) to rotate the mirror image of the arc to the side or completely out of the beam path. The arc’s focused image remains visible (Fig. 50).

• Use the adjusting buttons (46.1) and (46.5) to place the direct arc image right or left on an imaginary center line of the centering plane (Fig. 51).

• Then rotate the mirror image of the arc with the adjusting knobs (46.2) and (46.4), and focus it using the reflector (46.3).

• Orient the mirror image symmetrically to the direct image (Fig. 52). To do so, again use the adjusting knobs (46.2) and (46.4).

Fig. 50 Direct arc image focused but not centered

(in reality, the image is less focused)

Fig. 51 Direct arc image in target position

(in reality, the image is less focused)

• Using the collector, defocus the image with the collector head (46.6) until the arc image and mirror image are no longer recognizable and the image is uniformly illuminated.

• Replace the lamp adjustment reflector with the original filter cube.

Fig. 52 Direct arc image and mirror image in target

position (in reality, the image is less focused)

Page 49

7. Startup

Centering the Hg 100 W and Xe 75 W mercury lamps

• The adjustment window shows the direct image of the arc and its mirror image. These are generally not in alignment with one another.

• Focus the direct image with the collector (46.6).

• Use the adjusting buttons to pivot the arc’s mirror image on the rear side of the lamp housing (46.2, 46.4) to the side or completely out of the beam path. The arc’s focused image remains visible (Fig. 53).

• Use the adjusting buttons (46.1) and (46.5) to place the direct arc image in the middle of the centering plane, whereby the bright tip of the arc, the focal spot, should lie slightly outside of center (Fig. 54).

• Then pivot the arc’s mirror image with the adjusting knobs (46.2) and (46.4), and focus it using the reflector (46.3).

Fig. 53 Direct arc image focused but not centered

(in reality, the image is less focused)

Fig. 54 Direct arc image in target position

(in reality, the image is less focused)

• Orient the mirror image symmetrically to the direct image (Fig. 55). To do so, again use the adjusting knobs (46.2) and (46.4). The V-shaped irradiation of the direct image and mirror image arcs can be superimposed.

Caution!

The bright tips of the arcs, the focal spot, must never be projected onto each other, as this results in a danger of explosion by overheating.

Fig. 55 Direct arc image and mirror image in target

position (in reality, the image is less focused)

Page 50

7. Startup

Caution!

In older lamps, the structure of the arc is no longer clearly recognizable. The image is then more like that of a HG 50 lamp. The image and mirror image can no longer be superimposed exactly. In this case, align both images.

• Using the collector, defocus the image with the knob (46.6) until the arc image and mirror image are no longer recognizable and the image is homogeneously illuminated.

• Replace the lamp adjustment reflector with the original filter cube.

Note:

Make sure to switch off the instrument.

Page 51

8. Operation

8.1 Switching on the unit

When using a gas discharge lamp, the external supply unit must be turned on separately (56.1). Switch on the microscope or the CTR5000 at the power switch. All motorized microscope components will then run through an initialization phase. After the initialization is complete, the display on the stand (Fig. 57) shows the current microscope setting.

Fig. 56 Front view of the ebq 100 supply unit 1 Power switch 2 Lamp status

1 2

8.2 Stages and object displacement

Lengthening the coaxial drive

• For lengthening, pull the lower handle (58.2) downwards. Repeat with the upper handle (58.1).

Setting the movement rate (torque)

The torque for x and y can be individually adjusted using two knurled rings (58.3, 58.4).

Fig. 58 Rotating specimen stage 1 Object displacement (Y direction) 2 Object displacement (X direction) 3 Torque adjuster (Y-direction) 4 Torque adjuster (X-direction) 5 Focus wheel for fine focusing

Fig. 57 Display after initialization

1 2

Page 52

8. Operation

Rotating the stage

The swivel range of the rotating stages is 0° - 110°.

• Rotate the stage to loosen the locking screw (59b.1).

• Bring the table into the desired position.

• Retighten the locking screw.

Pol rotating stage (360°)*, Pol attachable mechanical stage *

The specimen can be fastened using either two spring-back stage clips or the Pol 3 multi-format attachable mechanical stage (Fig. 59a). For glass slides of approx. 26 mm (1") in width, the metal plate (59a.2) needs to be pivoted outwards and the specimen needs to be positioned according to Fig. 59a. If commercially available glass slides of 26 mm in width are placed perpendicular to that, then the range of movement of the attachable mechanical stage, which is normally approx. 30 mm x 40 mm, is not fully used. The pair of lock buttons provided allow for locking distances of 0.1, 0.3, 0.5, 1 and 2 mm. Changing them requires forceful, axial removal. When attaching the new lock button, make sure that the catch pins on the inside are properly positioned. For smaller microscopes, the limit stop screw on the underside must be shifted approx. 2 mm inwards toward the stroke limit. The two verniers allow the angular measurements to be read with a precision of 0.1.

45°click stop:

• Screw in the rotary knob (59a.5) until you can feel it starting to resist.

• Then turn the specimen stage to the next click stop.

• Loosen the rotary knob and look for the starting point of the next click stop (e.g., the object darkness setting).

• Retighten the rotary knob.

Now the rotating stage can be turned using 45° click stop intervals.

Fig. 59a Pol rotating stage* and Pol 3 attachable mechanical stage*

1 Holes for the fastening screw. 2 Lever for the holder for glass slides of various formats,

which can be turned inward and outward

3 Storage for the centering key 4 Locking button pair 5 45° click stop 6 Clamping system for the stage rotation

Page 53

8. Operation

8.3 Focusing

There is a focus wheel on the left side of the stand for coarse and fine focus adjustment (Fig. 59b).

On the right side of the stand, there is also a focus wheel, which is used exclusively for fine focusing (58.4). The special form of this handwheel makes it possible to simultaneously grasp the coaxial drive with your hand while operating the fine adjustment with one finger.

8.4 Tubes

Note:

Close any unused tube openings, as otherwise stray light can interfere with observation.

Note:

Make sure that the connecting cable is plugged in on the MBDT25+ motorized tube (60.1).

Adjust the interpupillary distance

• Adjust the interpupillary distance of the eyepiece tubes so that a congruent total image is seen (Fig. 60).

Fig. 59b Rotating specimen stage 1 Locking screw 2 Fine focusing 3 Coarse focusing

Fig. 60 Tube setting

↔ Setting the personal interpupillary distance

1 Motorized tube connection

↔

Page 54

8. Operation

Adjusting the viewing angle

• For the AET22 and EDT22 ergo tubes, the viewing angle can be adjusted by tilting the binocular eyepiece in the range of 5° – 32° (Fig. 61).

Adjusting the eyepiece extension to the arm length

• On the AET22 tube, the eyepieces can be extended up to 30 mm (Fig. 61).

Beamsplitting in photo tubes

EDT22 tube: The light division between the observation and documentation ports has a definite presetting (50:50).

BDT25+ tube: The light division is set manually by pulling out a control bar.

Control Bar Observation Photo VIS 100 % 0 % 50/50 150 % 50 % PHOTO 110 % 100 %

MBDT25+ tube: This tube is similar to the documentation tube BDT25+, but it is motorized. The control positions are selected using a variable function key on the stand.

HC L 2TU tube: The light division is set manually by pulling out a control bar.

Control Bar Observation Photo VIS 100 % 0 % PHOTO 110 % 100 %

Fig. 61 Individual settings of the AET22 tube

Fig. 62 BDT25+ tube with digital camera 1 Control bar

Page 55

8. Operation

8.5 Eyepieces

Note:

The eyepiece’s aperture protector must be removed or folded back, during microscopy while wearing glasses. We recommend removing bifocals and spectacles with progressive-addition lenses when using the microscope.

• For the adjustable tubes with documentation output, choose the 100% VIS position.

Eyepieces with inlaid reticle

• Focus the reticle by adjusting the eyelens in the eyepiece.

• Focus on the object through this eyepiece.

• Then, close that eye and focus on the object by adjusting the second ocular only.

8.6 Objectives

The objectives are moved into the beam path manually. Be sure that the turret locks into place.

The positions of the objectives in the objective turret have been specified at the factory and must be observed when installing the objectives. (See Installing objectives → p. 23)

When pivoting an objective inwards, the micro-

automatically selects:

scope

• The optimum setting for the field diaphragm

• The optimum setting for the aperture diaphragm and the light intensity for each contrast method.

The objective magnification and the total magnification appear in the display → p.39.

• Begin with a small level of magnification. Then switch to the next higher objective.

Correction for vision problems

• With your right eye, look through the right eyepiece and bring the specimen into sharp focus.

• Then, with your left eye, view the same position of the specimen and rotate the left eyepiece tube until this position is brought into sharp focus. Do not use the focus wheel.

• For immersion objectives use the appropriate

immersion medium. OIL: use optical immersion oil only

according to DIN/ISO standards.

Cleaning → p.78. W: Water immersion. IMM: Universal objective for water, glycerol,

oil immersion.

Caution!

Follow the safety instructions for immersion oil!

Page 56

8. Operation

For lockable immersion objectives:

• Lock these by pushing the front part upwards until it stops (approx. 2 mm).

• Then, after a gentle turning motion to the right, the objective is locked (Fig. 63).

For objectives with corrective mounts:

• Turn the knurled ring to adjust the objective to the thickness of the cover glass.

Objective centering * (DM4500 P)

Caution:

When changing the objective, the centering key must no longer be in the corresponding openings.

When centering the objectives (Fig. 64, 65), use two hexagon wrenches to move the objectives so that the optical axis of the objective (and, therefore, the center of the image) is aligned with the axis of rotation of the objective stage. If the objectives are centered correctly, a programmed specimen position will not drift out of the field of vision when the stage is turned. Therefore, a specimen point located inside the center of the cross-hairs does not change its position when the stage is rotated a full turn. When centering objectives, it is best to use a detailed, high-contrast specimen.

Fig. 63a Immersion objective, released Fig. 63b Immersion objective, locked

↔

Page 57

8. Operation

• Switch off the analyzer, the 1.6x tube lens and the Bertrand lens.

• Reduce the aperture diaphragm so that it is very small.

• Insert both objective centering keys above the objectives that need to be centered.

• Focus the specimen.

Two resembling methods can be used for centering objectives:

Method I (Fig. 64)

• Turn the specimen stage and note the position of the specimen that does not move in a circular path. This position of the specimen corresponds to the mechanical axis of rotation of the specimen stage.

• Now move the marked specimen position by shifting the two centering keys to the center of the cross-hairs.

• Turn the specimen stage and refine the centering as needed.

Method II (Fig. 65)

• Move the marked specimen position (65a) into the center of the M cross-hairs.

• Turn the specimen stage until the specimen position is as far from the center of the M cross-hairs as possible (Position A, Fig. 65b). In extreme cases, point A (= maximum deviation of the specimen position) can also be located outside of the field of view.

• Move the image by turning the centering key until position A of the specimen is located in the center (= Pos. B) between pos. A and the center of the M cross-hairs (65c).

• Move position A of the specimen to M and check to see whether A remains in M when the stage is rotated (65d). If necessary, repeat the centering procedure.

The objective centering procedure needs to be repeated for each objective. This ensures that the objectives retain their approximate centering settings when they are removed for cleaning, or other such procedures, and then reinserted into the same holes. If the coarse drive or the height adjustment device is used to change the height of the specimen stage (for example, when viewing thick specimens) the centering precision for all of the objectives may be reduced slightly.

Fig. 64 Centering method I

Fig. 65 Centering method II

M M

abcd

Page 58

8. Operation

8.7 Magnification changer

Optionally, a coded magnification changer can be used, which is manually operated. On the knurled ring, the following magnification factors can be set:

B stand M stand

1x 1x

1.25x 1.5x

1.6x 2x

The selected factor is indicated in the display and included in the total magnification.

8.8 HC P 1x/1.6x tube optics

with Bertrand lens (encoded)

These optics were developed especially for polarizing microscopy; however, they can also be used for all other methods.

• If needed, switch the Bertrand lens off and the 1x tube factor on. For the HC P (Pol) tube optics, switching to the 1x tube factor is sufficient.

Setting the tubes and eyepieces → p. 53f.

Features:

• 1x tube factor, can be switched to 1.6x

• Bertrand lens, can be activated, encoded, focused and centered

• Iris diaphragm in the intermediate image for masking out small particles (15 µm for the 100x objective).

Built-in, depolarized quartz plate:

This plate prevents interference colors, which are caused by tube prism polarization effects (pseudopleochroism), when the analyzer is connected and the polarizer is switched on; however, it only prevents this when the 1x tube factor is being used.

When using the 1.6 fix tube factor, make sure that the useful magnification for high objective magnifications and apertures (objective aperture x 1000) is not exceeded to the point that overmagnification causes a blurry image impression.

Page 59

8. Operation

8.9 Light sources

• Adjust the brightness using the function keys

(66.5). The INT function buttons are always assigned to the currently active transmitted light (TL) or incident light (IL) axis.

• For TL and IL: The setting can be made in coarse and fine steps. Pressing both INT buttons at the same time toggles between coarse and fine adjustment. The light intensity in the display changes accordingly.

0-20

Coarse adjustment:

======

0-255

Fine adjustment:

----------

• The intensity is individually adjusted and stored for each objective and contrast method.

• For fluorescence: The brightness is adjusted in 5 defined increments (FIM): 100% / 55% / 35% / 20% / 10%

8.10 Aperture diaphragm and field diaphragm

Both diaphragms have been set to suitable values for the current objective and contrast method at the factory.

• The diaphragms can be adjusted at any time

using AP (aperture diaphragm) (66.2) and FD (field diaphragm) (66.4) function keys.

Caution!

The old values will be overwritten by the current ones!

• The function keys are assigned to the currently active transmitted light (TL) or incident light (IL) axis.

Caution!

When the PH or DF is being used, the aperture diaphragm is opened completely and

cannot be

closed to prevent incorrect operations.

Fig. 66 Control panel 1 Variable function keys

(also on the right side of the stand)

2 Aperture diaphragm 3 Transmitted light / incident light 4 Field diaphragm 5 Light intensity

12345

Page 60

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

9.1 Transmitted light

9.1.1 Bright field (TL)

• Switch to the transmitted light axis (TL) by

pushing the TL/IL function key.

• Select the BF (bright field) contrast method.

Do so by pressing the BF variable key. Alternatively: press the variable button

CHANGE TL

(For details on key assignments, please see the identification sheet.) The display indicates BF.

• Insert a transmitted light specimen.

• Rotate an appropriate objective into place.

• Use the focus wheel to bring the image into

focus and set the brightness using the INT function key.

9.1.2 Phase contrast (TL)

• Switch to the transmitted light axis (TL) by

pushing the TL/IL function key.

• Select the PH (phase contrast) contrast method. Do so by pressing the variable button PH. Alternatively: press the variable button

CHANGE TL

(For details on key assignments, please see the identification sheet.) The display indicates PH.

• Insert a transmitted light specimen.

• Rotate an appropriate objective into place. Objectives that are suitable for phase contrast are engraved with PH.

• Bring the image into focus using the focus wheel and set the brightness using the INT function key.

Notes:

The microscope automatically selects the correct light ring in the condenser.

When selecting the phase contrast method, the aperture diaphragm is opened fully and can not be adjusted.

Page 61

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

9.1.3 Dark field (TL)

• Switch to the transmitted light axis (TL) by

pushing the TL/IL function key.

• Select the DF (dark field) contrast method by

pressing the variable button DF. Alternatively: press the variable button CHANGE TL (For details on key assignments, please see the identification sheet.) The display indicates DF. The dark field ring (dark field stop) is set automatically in the condenser.

• Insert a transmitted light specimen.

• Rotate an appropriate objective into place.

• Bring the image into focus using the focus

wheel and set the brightness using the INT function key.

The maximum objective aperture that can be used for dark field is 0.75. All objectives with greater apertures are automatically blocked for this method ("DF" flashes in the display).

Notes:

9.1.4 Polarization (TL)

• Switch to the transmitted light axis (TL) by pushing the TL/IL function key.

• Select the POL (polarization) contrast method by pressing the variable button POL. Alternatively: press the variable button

CHANGE TL

(For details on key assignments, please see the identification sheet.) The display indicates POL.

9.1.4.1 Manual method

• Rotate the polarizer on the underside of the condenser into the beam path (Fig. 67a). Make sure that the red index point on the front of the polarizer is at 0.

• Insert the analyzer up to the stop into the left side of the stand (Fig. 67b.1).

• Bring the polarizer and analyzer into cross position until they reach maximum darkness.

• Insert a specimen and rotate a suitable objective into place.

The microscope automatically selects the correct light ring in the condenser.

When the dark field method is selected, the aperture diaphragm is opened fully and cannot be adjusted.

Page 62

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

9.1.4.2 DM4500 P - examinations in polarized transmitted light

One polarizer only

If specimens need to be examined using other transmitted light methods, such as bright field, phase contrast and dark field, instead of with polarizers, then it is usually sufficient to switch off the analyzer or polarizer. If the brightness of the image is not adequate, the polarizer and the analyzer need to be switched off. Stained, birefringent specimens may exhibit brightness or color fluctuations when the specimen stage or the polarizer is rotated (with the analyzer switched off). This is called dichroism or pleochroism, which is an important indication when examining crystals. However, this effect can be simulated by nonpolarizing microscopes, because they do not contain depolarized quartz plates, or when an incident light reflector is left in the beam path during the transmitted light method. This also applies to using the 1.6x tube lens in the DM4500 P.

Fig. 67 Crossing the polarizers for observation using the Bertrand lens, objective with high aperture, without specimen

a crossed precisely b not crossed precisely

If pressure is present in the condenser or in the objective, position "a" cannot be set at all

Crossed polarizers

According to DIN and ISO ratings the run of the vibration directions corresponds to the table on p. 63. However, when polarizers are crossed, these vibrations have the same polarized optical appearances as they do when each of the polarizers are interchanged by 90°. If the specimen contains a large amount of nonbirefringent or opaque particles, then the analyzer is usually turned a few degrees away from the cross position to make these particles somewhat visible (for specimens that remain dark when the polarizers are crossed precisely). Examinations are not usually performed when the polarizers are parallel because this configuration is not sensitive enough to detect birefringent elements.

Changing the brightness when rotating birefringent objectives

When the specimen stage is rotated, the brightness of birefringent (anisotropic) specimens changes periodically. When the specimen is rotated, a total of 4 extinction positions (also called normal position) occur at each 90° interval. Exactly 45° between 2 extinction positions, 4 orientations of maximum intensity occur, the diagonal or 45° positions. In extinction, the specimen vibration directions are parallel to the transmission directions of the polarizers; at maximum intensity, the specimen vibration directions represent the bisectors of the polarizer directions. The cross-hairs in the (right) eyepiece on the polarizing microscope can be rotated N – S/E – W; therefore, they can be rotated in the direction of polarization or 45°, or set up according to the specimen vibration directions when in a diagonal position.

Page 63

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

Simple overview observations

• Place the transmitted light specimen on the

polarizer.

• Turn the condenser head inward.

• Focus through the condenser using a low-

power magnifying lens, such as a 5x.

Although this method does not provide good imaging performance; it does make it possible to view rows of specimen very quickly.

λ/4- and λ plate, quartz wedge

Depending on their design, λ/4 and λ plates are installed on the underside of the condenser or, for polarizing microscope, in the 8x condenser

∩

disk (the vibration direction γ runs: ) or they are inserted in the tube slot. An automatic, spring-mounted dust cover flap closes the tube slot. For the IC/P analyzer, the λ plate can be activat- ed by turning it so that the mark "I" is facing upwards. When the plate is activated, the path difference is increased or reduced according to Fig. 68. The corresponding color changes can be used to determine the vibration direction γ according to the larger refractive index (i.e. refractive index n

). (The quartz wedge (69.7) allows

various color shifts on the polarizing microscope.)

Circular polarization

When the specimen stage is rotated, birefringent specimens show 4 extinction positions. When a larger number of birefringent specimens are present, a few birefringent specimens always show up randomly in the extinction position. Circular polarization is used for simultaneous interference color observation of all specimens:

• Remove the specimen from the beam path or locate the blank position of the specimen.

• Cross the polarizers precisely. The polarizers must be positioned exactly in the N – S/E – W direction. This means that the analyzer must either be set exactly at 90° or 0° of polarization.

Fig. 68 Interference color with dependence on the path difference or on the thickness and color changes in the addition and subtraction areas of the λ and λ/4 plates.

schwarz lavendelgrau

– λ

+ λ

graublau gelblichweiß

lebhaftgelb

rotorange tiefrot indigo himmelblau

grünlichblau hellgrün reingelb

orangerot dunkelviolettrot

indigo grünlichblau

meergrün grünlichgelb

fleischfarben kaminrot

mattpurpur

λ

– – 4

λ

+ –

200

1. Ordnung

400

600

800

2. Ordnung

1000

1200

Gangunterschied

1400

3. Ordnung

1600

Page 64

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

• Insert the λ/4 plate (69.5) into the tube slot.

• Insert the λ/4 plate (69.1) into the receptacle located on top of the polarizer and rotate it until a specimen-free field of vision appears at its maximum extinction position (make sure to cross the polarizers precisely before doing this).

Compensators for quantitative measurements

Adjustable compensators are used to obtain exact measurements of path differences. To obtain the usual specimen thickness, d, and the measured path difference, gamma (Γ), the birefringence ∆n’ is calculated according to the following formula:

Γ = d x ∆n’ [nm] or ∆n = ––

When obtaining measurements, the compensator is inserted into the compensator slot and adjusted until the specimen position being measured is at the maximum extinction position. For this, the specimen must be pitched at a diagonal.

For HC P tube optics, use an iris diaphragm to mask out the measuring points. Detailed information can be found in the instructions for the compensator.

The following compensators are available:

Elliptical compensator based on Brace-Köhler techniques 69.9) Rotating compensator with compensator plate and a path difference of approx. λ/10. Measurements can be obtained in white or monochromatic light with a measurement range of up to approx. 50 nm.

Elliptical compensator based on Sénarmont techniques 69.6) (λ/4 plate in subparallel position) Measurements are obtained in monochromatic light (546 nm), which requires an analyzer that can be rotated 360°. In most cases, the compensator is used to measure path differences of up to 1 order. But higher path differences can be measured. However, when doing so, the compensator does not provide the entire path differ-

Fig. 69 Compensators 1, 2 λ/4 u. λplate. For polarizing microscopes only: 3 λ/4 u. λplate for the 8x turret disk, 4, 5 λ/4 u. λplate for the tube slot, 6 Revolving λ/4 plate (Sénarmont compensator), 7 Quartz wedge, 8 Tilt compensator, 9 Brace-Köhler compensator

Page 65

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

ence; rather, it provides only the difference over the entire wavelength or over a multiple of the wavelength. Entire wavelengths must be defined using a tilt compensator, a quartz wedge and by measuring the interference color. The results are more accurate than those obtained using a tilt compensator only.

Tilt compensator B with a measurement range of up to 5 orders based on Berek techniques Compensator (69.8) with an MgF

plate for mea-

suring up to approx. 5 orders of path differences in white or monochromatic light. You can read the path difference directly from the provided measurement table by calculating the sum of the two angles that are formed when the compensators are tilted simultaneously.

Tilt compensator K with a measurement range of up to 30 Orders (69.7) For measuring path differences in white or monochromatic light up to the specified maximum path difference. The compensator plate is made of calcite. The evaluation is created by performing simple calculations using the provided tables and the specified measured constants. Measure in white or monochromatic light.

Using conoscopy for crystal structures

Because the interference images appear in the eyepoint, they are not visible during typical observation (orthoscopy). An improvisational method for observing these images is to remove the eyepiece from the tube and to use a monocular, held a few cm away, for looking into the tube. You can improve the observation by using a focusing telescope for the phase contrast.

However, additional crystals located in the field of view need to be masked out because they disrupt the interference images of a crystal located in the middle of the field of view.

Setting the conoscopy

For conoscopy, the specimen positions that are most suitable are those that have the lowest path differences (table Fig. 68). Efficient conoscopic observation requires that the objective be centered precisely and that the cross position of the polarizers is exact.

• Rotate an objective with an aperture that is as high as possible (e.g. 40x, 50x or 63x) into the beam path.

• Rotate the condenser head into the beam path.

Birefringent crystals create interference images, also called axis images or conoscopic images (Fig. 71a,b), in the exit pupil of the objective (e.g. inside the objective). The form of these interference images and the changes that occur in these images when using a compensator make it possible to state how many axes the crystals have (uniaxial or biaxial crystals), how the axes are oriented, and whether or not the birefringence is positive or negative (positive or negative birefringent crystals).

• Open the aperture diaphragm.

• Place the crystal being examined as close to the center of the field of vision as possible.

• Turn the 1.6x tube lens inward.

• Widen or narrow the iris diaphragm according to the size of crystal, and make the field diaphragm narrower if necessary.

Page 66

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

• Insert the Bertrand lens (Fig. 70B) and focus it by rotating the operating button until the interference image or the circular diffuse edge of the eyepoint is in focus. If needed, center the Bertrand lens: Insert the hexagonal keys into the centering holes in sequence. If necessary, align the right eyepieces so that the cross-hairs correspond approximately to the direction of movement during the centering process.

• Adjust the collector to its optimum setting; use the diffuser if necessary.

Determining the optical character

Uniaxial crystals (Fig. 71a) When observing uniaxial crystals in the conoscopic (diverging) beam path, a dark cross appears whose center point indicates the optical axis. The cross is surrounded by colored interference bands*. When a variable compensator (quartz wedge or tilt compensator) is used, the rings move toward the center point or outside two opposite quadrants in the cross. The optical character results from the direction of the movement of the rings according to Fig. 71. Cross sections in which the crystal optical axis is sloped toward the direction of the viewer are suitable for determining the optical character. In addition, an optical character can be determined even if the center point of the cross is located outside of the field of view. Fig. 71 shows that fixed compensators can be used in place of variable compensators for determining the optical character.

Even if only one of the optical axes is within the observer’s direction of sight, the optical character can usually be identified. The brightness for specimens oriented in this way changes in the orthoscopic beam path very little or not at all when the objective is rotated. Consequently, only one of the two isogyres are visible in the conoscopic beam path.

Biaxial crystals (Fig. 71b) The cross sections in which the bisectors of the optical axes run parallel to the direction of sight are especially suitable for determining the optical character (the section is perpendicular to the acute bisectrix). A dark cross can be identified in the divergent beam path. This cross splits into two hyperbolic lines, also called isogyres, when the specimen stage is turned. The cross and or hyperbolic isogyres are surrounded by colored interference bands. After the compensator has been activated, the optical character can be determined from the direction of movement of these bands according to Fig. 71 or the following rule: The screw axis symmetry of the isogyres must run perpendicular to the γ−direction of the compensator:

Fig. 70 Functions of the HC P Pol tube optics

Control element 1x Orthoscopy 1.6x Orthoscopy Conoscopy

Tube lens 1x 1.6x 1.6x Iris diaphragm user-defined, adapted for > Specimen

Bertrand lens in the beam path off on Polarization on or off on or off crossed

as it is not the field of vision

(not for Dichroism/ Pleochroism)

O/B

1.6x

Page 67

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

Biaxial, positive crystals: When the compensator is activated, the movement of the interference bands proceeds from the convex side to the concave side of the isogyres.

Biaxial, negative crystals: The interference bands move from the concave side to the convex side.

Possible faults

The polarizers have been damaged (discolored) by strong light sources or they are heavily soiled.

The objectives or the condenser have been deformed by mechanical damage.

There is a beam splitter or filter between the polarizers.

The embedding material for transmitted light specimens is birefringent.

Fig. 71a Determining the optical characters for uniaxial structures Left: Positive uniaxial crystal, cut perpendicular to the optical axis. Right: Negative uniaxial crystal, cut perpendicular to the optical axis.

1 Illustration of the vibration directions in the specimen and in the compensator 2 Interference figure changes when using a λ/4 plate, 3 Interference figure changes when using a λ/plate,

Fig. 71b Table for determining the optical character

Orientation

of the

condenser

plate

ubu

ssch

itz

bleu

blue

blau

jaune

yellow

gelb

blau

blue

gelb

bleu

yellow

jaune

–

jaune

yellow

bleu

gelb

blue

blau

gelb

yellow

jaune

blau

blue

bleu

–

* When using 1/4-λ glimmer plates, black points appear instead of black arcs.

Uniaxial Biaxial

–

Page 68

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

9.1.4.3 Motorized method

• After the POL contrast method has been se- lected, the position of the polarizer is switched inside the condenser (if the microscope is equipped with these components). The analyzer cube is also automatically brought into the beam path.

9.1.4.4 Combined methods

• For the Leica DM4000 B, DM4500 P and Leica DM5000 B microscopes, it is possible to combine purely mechanical and motorized components.

For DM4500 P only:

After the Bertrand lens (links) is turned inwards, CONOS appears on the display. The light intensi-

ty and aperture and field diaphragms are assigned to the objective currently in use (objective magnification for conoscopy is 40x, 63x or 100x).

9.1.5 Differential interference contrast (TL)

9.1.5.1 DM4500 P

• Switch to the transmitted light axis (TL) by

pushing the TL/IL function key.

• Insert a specimen and rotate a suitable objective into place.

• Select the DIC contrast method. Do so by pressing the DIC variable key. Or by pressing the variable button. CHANGE TL

. (For details on key assignments, please see the identification sheet.) ICT appears on the display.

• Turn the polarizer on the underside of the condenser into the light path. Make sure that the red index point on the front of the polarizer is at 0 (Fig. 72).

Fig. 72 Turn the polarizer inward 1 Polarizer

Fig. 73 Inserting the analyzer 1 Analyzer

Page 69

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

• Insert the analyzer on the left side of the stand up to the click stop (fig. 73).

• Then insert the objective prism slider into the slot on the objective turret.

• The DIC prism that needs to be used appears on the display.

• For fine adjustment, rotate the knurled screw (74.1) on the objective prism slide.

9.1.5.2 DM5000 B

• Switch to the transmitted light axis (TL) by pushing the TL/IL function key.

• Apply a specimen and rotate a suitable objective into place.

• Select the DIC contrast method. Do so by pressing the DIC variable key. Alternatively: press the variable button CHANGE TL

. (For details on key assignments, please see the identification sheet.) ICT appears on the display.

Alternatively:

• Manually rotate the polarizer on the underside of the condenser into the beam path (Fig. 72).

• Likewise, manually insert the analyzer up to the stop into the left side of the stand (Fig. 73). The objective and condenser prisms are automatically moved into the beam path.

• The polarizer located in the condenser and the fitting condenser prisms are automatically brought into the beam path. The corresponding objective prism and the analyzer cube are also positioned automatically.

• For fine adjustment, use the knurled wheel above the objective turret.

Fig. 74 Motorized variants 1 Knurled wheel for fine focusing

Page 70

9. Contrast methods for Leica DM4000 B/DM4500 P/DM5000 B

• For fine adjustment, use the knurled wheel above the objective turret.

9.2 Fluorescence

• Switch to the fluorescence axis / fluorescence cube (FLUO) by pushing the TL/IL function key.

• Apply a specimen and rotate a suitable objective into place.

• The current fluorescence filter cube is indicated on the display.

• You may protect your specimen from fading by closing the incident light shutter. Do so by pressing the SHUTTER variable button. (For details on key assignments, please see the identification sheet.)

↓

This symbol appears on the display:

• Changing the fluorescence filter cube: By pressing the variable key

Cube

or Cube

• Bring the image into focus using the focus

wheel and set the brightness using the INT function key.

• The fluorescence intensity can be increased using the booster lens on the right side of the stand (Fig. 75). Leica recommends that you insert the booster lens into the front slot.

• For multifluorescence, use of an excitation manager is recommended. The excitation manager is inserted into the stand up to the last click stop on the right side of the stand (Fig. 76). Leica recommends that you insert the excitation manager into the front slot.

Fig. 75 Inserting the booster lens

Fig. 76 Inserting the excitation manager

Page 71

10. Contrast methods for Leica DM4000 M

10.1 Incident light

10.1.1 Bright field (IL)

• Switch to the incident light axis (IL) by push-

ing the TL/IL function key.

• Select the BF (bright field) contrast method

Do so by pressing the BF variable key. Alternatively: press the variable button

CHANGE RL

(For details on key assignments, please see the identification sheet.) The display indicates BF.

• Insert a specimen.

• Rotate an appropriate objective into place.

• Bring the image into focus using the focus

wheel and set the brightness using the INT function key. The brightness values for each objective are saved.

10.1.2 Dark field (IL)

• Switch to the incident light axis (IL) by push-

ing the TL/IL function key.

• Select the DF (dark field) contrast method

by pressing the variable button DF. Alternatively: press the variable button

CHANGE RL

(For details on key assignments, please see the identification sheet.) The display indicates DF. The DF reflector is rotated into the beam path.

• Insert a specimen.

• Rotate an appropriate objective into place.

• Bring the image into focus using the focus

wheel and set the brightness using the INT function key.

The maximum objective aperture that can be used for dark field is 0.75. All objectives with greater aperture are automatically blocked for this method ("DF" flashes in the display).

Notes:

When the dark field method is selected, the aperture diaphragm is opened fully and cannot be adjusted.

Page 72

10. Contrast methods for Leica DM4000 M

10.1.3 Polarization (IL)

• Switch to the incident light axis (IL) by push-

ing the TL/IL function key.

• Select the POL (polarization) contrast method

by pressing the variable button POL. Alternatively: press the variable button CHANGE RL

. (For details on key assignments, please see the identification sheet.) The display indicates POL.

Automatic procedure:

• The ICR filter cube is automatically brought into the beam path.

Manual procedure:

• Manually push the corresponding polarizer (77.3) and the IC/P analyzer (78.1) on the stand into the beam path. Then bring the polarizer and analyzer into cross position until they reach maximum darkness.

• Apply a specimen and rotate an objective with a low magnification level into place.

Fig. 77 Objective prism slide 1 Knurled wheel for fine focusing 2 Prism slot with inserted prism slide bar 3 Insert the polarizer

Fig. 78 1 Insert the analyzer

Page 73

10. Contrast methods for Leica DM4000 M

10.1.4 Interference contrast (RL)

• Switch to the incident light axis (IL) by push-

ing the TL/IL function key.

• Insert a specimen and rotate a suitable objective into place.

• Select the DIC contrast method Do so by pressing the DIC variable key. Alternatively: press the variable button

CHANGE RL

(For details on key assignments, please see the identification sheet.) The display indicates ICR.

• The ICR filter cube (containing the polarizer and analyzer) is automatically brought into the beam path on the incident light axis. Insert the objective prism slide into the prism slot (Fig. 77.2).

Alternatively:

• Manually push the ICR polarizer (77.3) and the IC/P analyzer (78.1) on the stand into the beam path.

• Then insert the objective prism slider into the slot on the objective turret. (77.2).

• For fine adjustment, rotate the knurled screw (77.1) on the objective prism slide.

10.2 Transmitted light

10.2.1 Bright field (TL)

• Switch to the transmitted light axis (TL) by

pushing the TL/IL function key.

• Select the BF (bright field) contrast method Do so by pressing the BF TL variable key. (For details on key assignments, please see the identification sheet.) The display indicates BF TL.

• Insert a transmitted light specimen.

• Rotate an appropriate objective into place.

• Bring the image into focus using the focus wheel and set the brightness using the INT function key.

10.2.2 Polarization (TL)

• Switch to the transmitted light axis (TL) by pushing the TL/IL function key.

• Select the POL (polarization) contrast method.

• Push the analyzer on the left side of the stand into the beam path (Fig. 78).

• Turn the polarizer on the underside of the condenser into the light path. Make sure that the red index point on the front of the polarizer is at 0 (Fig. 72).

Page 74

11. Troubleshooting

Problem

Stand

The microscope does not respond.

Illumination

The image is completely dark.

Cause/remedy

Ensure that the AC outlet has power. Make sure that the microscope is connected

to the power supply.

Check the cable connections. Inform Service and have the supply unit fuse

checked.

Open the shutter (→ Check the connection to the lamp housing on

p. 39).

the microscope. Transmitted light connection: Incident light (fluorescence) connection:

Make sure that the lamps are connected to

the power supply.

Inform Service and have the ebq 100 supply

unit fuse checked.

The image is unevenly or not uniformly illuminated.

The illumination flickers.

The lamp does not illuminate immediately upon being switched on.

Remove all unneeded filters from the light

path.

Center the lamp (→ Replace the old lamp (→

Be sure that there is no loose connection at

p. 47ff).

p. 23ff).

the power supply.

Replace the old lamp (→

The ebq 100 must be switched-on repeatedly. Hot Hg lamps should cool down before being

p. 23ff).

switching on again.

Page 75

11. Troubleshooting

Problem

Bright field

The specimen can not be brought into focus.

Dark field

No definite DF contrast is possible.

The image is unevenly or not uniformly illuminated.

Cause/remedy

Use the correct immersion medium. Lay the specimen with the cover glass toward

the top.

Make sure that the cover glass thickness is

correct and that it meets the specifications on the objective.

Be sure that a DF objective is being used. The objective aperture setting is too high

(maximum 0.75). If necessary, reduce the objective aperture using the iris diaphragm on the objective.

Check the condenser centering.

The magnification is too weak.

Use a higher magnification.

Undesirable stray light.

Phase contrast

No phase contrast is possible.

Clean the specimen and neighboring lens sur-

faces (→

The specimen is too thick. The cover glass is not placed planar. Check the centering of the light rings

p. 77).

(→ p. 44).

Page 76

11. Troubleshooting

Problem

Polarization

No polarization contrast is possible.

The image is insufficient.

The image is too dark.

No conoscopy image is possible.

Fluorescence

Cause/remedy

Bring the polarizer and analyzer into cross po-

sition until they reach maximum darkness (without specimen).(→ p. 61ff, 72).

The embedding material for transmitted light

specimen is birefringent.

Polarizers can be damaged by strong light

sources. Check to see if the polarizer is damaged (discolored) or heavily soiled.

Check to see if a beam splitter or filter is

switched on.

Objectives and condensers can be deformed

by mechanical damage.

The image is completely dark (no fluorescence).

The fluorescence is too weak.

Display

The display flashes.

FAIL! appears on the display.

Open the shutter (→ Select the incident light axis (IL) (→ Check the antigen-antibody combination.

Insert the booster (→ Center the lamp (→ Insert a new lamp (→

Iotate an appropriate objective for the con-

p. 70).

p. 33).

p. 47ff).

p. 23f).

p. 40).

trast method into place.

Check the installed objectives, filter cubes, etc. Switch the microscope off and back on.

Page 77

12. Care of the microscope

12.2 Cleaning

Caution!

12. Care of the microscope

Unplug the power supply before performing cleaning and maintenance work! Protect electrical components from moisture!

Microscopes in warm and warm-damp climatic zones require special care in order to prevent the build up of fungus. The microscope should be cleaned after each use, and the microscope optics should be kept strictly clean.

12.1 Dust cover

Note:

To protect against dust, cover the microscope and accessories with the dust cover after each use.

Caution!

Let lamps cool down before covering the stand with a dust cover. The dust cover is not heat-resistant. In addition condensation water may occur.

Caution:

Residual fiber and dust can create unwanted background fluorescence.

Cleaning coated parts

Dust and loose dirt particles can be removed with a soft brush or lint-free cotton cloth.

Stubborn dirt can be removed with all commonly available aqueous solutions, naphtha or alcohol. For cleaning coated parts, use a linen or leather cloth that is moistened with one of these substances.

Caution:

Thinners containing acetone, xylene or nitrogen can harm the microscope and thus must not be used.

Test cleaning solutions of unknown composition first on a less visible area of the unit. Be sure that coated or plastic surfaces do not become matted or etched.

Cleaning the stage

Rub the stage with paraffin oil or acid-free Vaseline to remove light spots on the stage.

Page 78

12. Care of the microscope

Cleaning glass surfaces

Remove dust on glass surfaces with a fine, dry and grease-free brush made from hair, by blowing with a squeeze blower or vacuum suction.

Remove stubborn dirt on glass surfaces with a clean cloth dampened with distilled water. If the dirt still can not be removed, use pure alcohol, chloroform or petroleum ether.

Cleaning objectives

Caution!

The objective may not be unscrewed during cleaning. If damage appears on inner surfaces, the objectives must be sent to your Leica subsidiary for repair. We also advise against cleaning the inside surfaces of the eyepieces.

The front lenses of objectives are cleaned as described under "Cleaning Glass Surfaces". The upper lens is cleaned by being blown off with a pneumatic pump.

Removing immersion oil

Caution!

Follow safety instructions for immersion oil!

First, wipe off the immersion oil with a clean cotton cloth, and then re-wipe the surface several times with ethyl alcohol.

12.3 Handling acids and bases

For examinations using acids or other aggressive chemicals, particular caution must be taken.

Caution:

Be absolutely certain to prevent the optics and mechanical parts from coming into contact with these chemicals.

Page 79

13. Essential wear and spare parts

Order No. Material No. Name Used for

Replacement lamps 11 500 974 Halogen bulb 12 V 100 W 107/2 lamp housing 11 500 137 High-pressure mercury burner 50W 106 z lamp housing 11 500 138 High-pressure mercury burner 100W 106z lamp housing 11 500 321 High-pressure mercury burner 100W 106z lamp housing

11 500 139 High-pressure xenon burner 75 W 106 z lamp housing

Screw cap for unused objective receptacles 020-422.570-000 Screw cap M 25 Objective turret

Replacement eyecup (antiglare protection) for HC PLAN eyepiece 021-500.017-005 HC PLAN eyecup 10x/25 eyepiece 021-264.520-018 HC PLAN eyecup 10x/22 eyepiece 021-264.520-018 HC PLAN eyecup 10x/20 eyepiece

Immersion oil 11 513 860 120 ml OIL and IMM objectives 11 513 861 250 ml and oil condenser heads 11 513 859 10 ml, without self-fluorescence

(103 W/2)

Page 80

14. Abbreviations and pictograms

Contrast method



Magnification

Light intensity / diaphragms

Light division in the motorized tube

↑

↓

AET Advanced ergo tube AP Aperture diaphragm BF Bright field COMBI Combination methods CONOS Conoscopy CUBE Fluorescence cube DF Dark field incident / transmitted light DIC Differential interference contrast FD Field diaphragm FLUO Fluorescence axis (incident light) ICR Interference contrast (incident light) ICT Interference contrast (transmitted light) IL Incident light INT Intensity MBDT Motorized basic documentation tube PH Phase contrast POL Polarization, incident / transmitted light RL Incident light TL Transmitted light

Transmitted light shutter is open

Transmitted light shutter is closed

Incident light shutter is open

Incident light shutter is closed

Page 81

15. Index

Adjusting the light sources 45

Allowable ambient conditions 18 Ambient conditions 18 Ambient temperature 10 Analyzer 32, 61, 69, 72 Analyzer cube 68, 69 Aperture diaphragm 15, 39, 59

Beam splitting in photo tubes 54 Bertrand lens 58, 68 Booster lens 70 Bright field 60, 71

Checking the phase contrast

rings 44 Circular polarization 63 Cleaning 77 Cleaning objectives 78 Coaxial drive 51 Compensators 64 Condenser 13, 15, 21 Condenser centering 41 Condenser connection 21 Condenser height adjuster 19, 21 Condenser holder 21 Conoscopy 65 Conoscopy module 13, 38 Contrast method 12, 60, 71 Correction for vision problems 55

Dark field 61, 71 Dark field stop 61 Defined function key 40 DIC prisms 32 Differential interference contrast 68 Display 15, 39 Disposal 11

ebq 100 supply unit 10, 29, 51 Electrical safety 10 ErgoModule 33 Excitation manager 33, 70 Eyepiece 15

Field diaphragm 15, 39, 59 Filter block changer 16 Filter cube 30 Filter cube ICR 72, 73 FIM (fluorescence intensity

manager) 12, 59

Fluorescence 70 Fluorescence intensity 70 Focus finder 37 Focusing 41, 53 Focusing telescope 44 Function buttons 35, 40

Gas discharge lamps 27, 28

Halogen lamp 26, 47

HC P tube optics 58, 66 Hg 100 W and Xe 75 W

mercury lamps 49 Hg 50 burner 28 Hg 50 W mercury lamp 48

Identification sheet

23, 30, 60, 61, 69, 70, 71, 72, 73 Illumination manager 12 Immersion oil 55, 78 Incident light axis 12 Incident light polarizers 31 Incident light shutter 39 Incident light turret disk 30 Initialization 38 Interference contrast 73 Intermediate systems 19

Köhler illumination 21, 41

λ/4 and λ plate 63

Lamp housing 16 Lamp housing 106 z 25, 46 Lamp housing 107/2 23, 45 Lamp housing

receptacle 24, 26, 29 LAS software package 13, 17 Leica CTR5000 electronics

box 10, 34 Leica SmartTouch 13 Light intensity 39 Light sources 45, 59 Light sources for the incident

light axis 25 Light sources for the transmitted light

axis 23 Locking pin 30

Magnification changer 13, 58 Mirror housing 33 Motorized analyzer 32 Motorized polarizer 31

Objective aperture 61 Objective centering 56 Objective prism slide 73 Objective turret 12, 15 Objectives 55 Operating buttons 15 Optical character 66

Phase contrast 44, 60 Pol attachable mechanical

stage 52 Polarization 61, 72 Polarizer 61, 62, 69, 72 Polarizer L/ICR 31 Polarizer R/ICR 31 Polarizer R/P 31 Power supply connection 34 Prism slot 73

Quartz plate 58 Quartz wedge 63

Reflector cube 30 Reflector cube for lamp

adjustment 45 Reset function 35 Rotating polarizer 31 Rotating stage 20, 52

Shutter 70 Specimen holder 19 Specimen stage 19

Transmitted light / incident

light toggle 15 Transmitted light and incident

light analyzer 32 Transmitted light axis 12 Transmitted light filter 16 Transmitted light polarizer ICT/P 31 Transmitted light shutter 39

Variable function

buttons 15, 16, 35, 40

Xe 75 burner 28

Page 82

16. EU Declaration of Conformity

Download:

http://www.light-microscopy.com/down_ce-declaration_DM4000b http://www.light-microscopy.com/down_ce-declaration_DM4000m http://www.light-microscopy.com/down_ce-declaration_DM4500p http://www.light-microscopy.com/down_ce-declaration_DM5000b

Page 83

Leica DM4000 B Leica DM4000 M Leica DM4500 P Leica DM5000 B

Bedienungsanleitung

Page 84

Copyrights

Alle Rechte an dieser Dokumentation liegen bei der Leica Microsystems CMS GmbH. Eine Vervielfältigung von Text und Abbildungen – auch von Teilen daraus – durch Druck, Fotokopie, Mikrofilm oder andere Verfahren, inklusive elektronischer Systeme, ist nur mit ausdrücklicher schriftlicher Genehmigung der Leica Microsystems CMS GmbH gestattet.

Der Begriff Windows kann im folgenden Text ohne weitere Kennzeichnung verwendet werden. Hierbei handelt es sich um ein geschütztes Warenzeichen der Firma Microsoft Corporation. Ansonsten kann aus der Verwendung von Warennamen ohne besondere Hinweise kein Rückschluss auf deren freie Verwendbarkeit gezogen werden.

Die in der folgenden Dokumentation enthaltenen Hinweise stellen den derzeit aktuellen Stand der Technik sowie den derzeit aktuellen Wissensstand dar. Die Zusammenstellung von Texten und Abbildungen haben wir mit größter Sorgfalt durchgeführt. Trotzdem kann für die Richtigkeit des Inhaltes dieses Handbuches keine Haftung irgendwelcher Art übernommen werden. Wir sind jedoch für Hinweise auf eventuell vorhandene Fehler jederzeit dankbar.

Die in diesem Handbuch enthaltenen Informationen können ohne vorherige Ankündigung geändert werden.

Page 85

Inhalt

1. Wichtige Hinweise zur Anleitung ......... 7

2. Zweckbestimmung der Mikroskope ..... 8

3. Sicherheitshinweise ................................ 9

3.1 Allgemeine Sicherheitshinweise ........... 9

3.2 Elektrische Sicherheit .............................. 10

3.3 Entsorgung.................................................. 11

4. Geräteübersicht ........................................ 12

5. Auspacken.................................................. 17

6. Montage des Mikroskops ....................... 19

6.1 Objekttisch .................................................. 19

6.2 Kondensor ................................................... 21

6.3 Tubus und Okulare..................................... 22

6.4 Objektive ..................................................... 23

6.5 Lichtquellen für die Durchlichtachse .... 23

6.6 Lichtquellen für die Auflichtachse ......... 25

6.7 Bestückung der

Auflicht-Revolverscheibe ........................ 30

6.8 Polarisator und Analysator...................... 31

6.9 DIC-Prismen ............................................... 32

6.10 Optionales Zubehör ................................... 33

6.11 Anschluss an die Stromversorgung ...... 34

6.12 Anschluss

an die Elektronikbox CTR5000 ................. 34

7. Inbetriebnahme ......................................... 35

7.1 Funktionsprinzip......................................... 35

7.2 Einschalten ................................................. 38

7.3 Das Display

(DM4000 B/4000 M/4500 P) ...................... 39

7.4 Die Funktionstasten .................................. 40

7.5 Köhlersche Beleuchtung ......................... 41

7.5.1 Durchlicht......................................... 41

7.5.2 Auflicht ............................................. 42

7.6 Phasenkontrastringe überprüfen ........... 44

7.7 Einstellung des motorischen

Polarisators (DM4500 P/DM5000 B)....... 45

7.8 Justieren der Lichtquellen....................... 45

8. Bedienung .................................................. 51

8.1 Einschalten ................................................. 51

8.2 Tische und Objektverschiebung............. 51

8.3 Fokussierung .............................................. 53

8.4 Tuben ........................................................... 53

8.5 Okulare ........................................................ 55

8.6 Objektive ..................................................... 55

8.7 Vergrößerungswechsler .......................... 58

8.8 Tubusoptik HC P 1x/1.6x ........................... 58

8.9 Lichtquellen ................................................ 59

8.10 Aperturblende und

Leuchtfeldblende....................................... 59

Page 86

Inhalt

9. Kontrastverfahren für Leica DM4000 B/DM4500 B/

DM4500 P/DM5000 B ................................ 60

9.1 Durchlicht ................................................... 60

9.1.1 Hellfeld ............................................... 60

9.1.2 Phasenkontrast................................. 60

9.1.3 Dunkelfeld.......................................... 61

9.1.4 Polarisation ....................................... 61

9.1.4.1 Manuelles Verfahren ................... 61

9.1.4.2 DM4500 P – Untersuchungen

im polarisierten Durchlicht ......... 62

9.1.4.3 Motorisches Verfahren ............... 68

9.1.4.4 Kombinierte Verfahren ................ 68

9.1.5 Differentieller

Interferenzkontrast ........................ 68

9.1.5.1 DM4500 B/DM4500 P .................... 68

9.1.5.2 DM5000 B........................................ 69

9.2 Fluoreszenz ................................................. 70

10. Kontrastverfahren für

Leica DM4000 M ........................................ 71

10.1 Auflicht ........................................................ 71

10.1.1 Hellfeld ............................................. 71

10.1.2 Dunkelfeld........................................ 71

10.1.3 Polarisation ..................................... 72

10.1.4 Interferenzkontrast ........................ 73

10.2 Durchlicht ..................................................... 73

10.2.1 Hellfeld ............................................. 73

10.2.2 Polarisation ..................................... 73

11. Trouble Shooting ....................................... 74

12. Pflege des Mikroskops............................ 77

12.1 Staubschutz ................................................ 77

12.2 Reinigung .................................................... 77

12.3 Umgang mit Säuren und Basen .............. 78

13. Wichtigste Verschleiß- und

Ersatzteile ................................................... 79

14. Abkürzungen und Piktogramme............. 80

15. Index ............................................................ 81

16. EU-Konformitätserklärung ...................... 82

Page 87

1. Wichtige Hinweise zur Anleitung

Achtung!

Diese Bedienungsanleitung ist ein wesentlicher Bestandteil des Mikroskops und muss vor Inbetriebnahme und Montage sorgfältig gelesen werden.

Textsymbole, Piktogramme und ihre Bedeutung:

(1.2)

→ S.20

Diese Bedienungsanleitung enthält wichtige Anweisungen und Informationen für die Betriebssicherheit und Instandhaltung des Mikroskops und der Zubehörteile. Sie muss daher sorgfältig aufbewahrt werden.

Ziffern in Klammern, z.B. (1.2), beziehen sich auf Abbildungen, im Beispiel Abb.1, Pos. 2.

Ziffern mit Hinweispfeil, z.B. → S.20, weisen auf eine bestimmte Seite dieser Anleitung hin.

Achtung! Besondere Sicherheitshinweise in dieser Anleitung sind durch das nebenstehende Dreieckssymbol gekennzeichnet und grau unterlegt.

Achtung! Bei einer Fehlbedienung können Mikroskop bzw. Zubehörteile beschädigt werden.

Erklärender Hinweis.

Hinweise zur Entsorgung von Mikroskop, Zubehörkomponenten und Verbrauchmaterial.

Nicht in allen Ausrüstungen enthaltene Position.

Page 88

2. Zweckbestimmung der Mikroskope

Die Mikroskope DM4000 – DM5000, zu denen diese Bedienungsanleitung gehört, und die den Kennbuchstaben B tragen, sind für biologische Routine- und Forschungsanwendungen vorgesehen. Dies schließt die Untersuchung von aus dem menschlichen Körper stammenden Proben zum Zwecke der Informationsgewinnung über physiologische oder pathologische Zustände oder angeborene Anomalien oder zur Prüfung auf Unbedenklichkeit und Verträglichkeit bei potenziellen Empfängern oder zur Überwachung therapeutischer Maßnahmen ein.

Für jegliche nicht-bestimmungsgemäße Verwendung und bei Verwendung außerhalb der Spezifikationen von Leica Microsystems CMS GmbH, sowie gegebenenfalls daraus entstehender Risiken übernimmt der Hersteller keine Haftung. In solchen Fällen verliert die Konformitätserklärung ihre Gültigkeit.

Achtung!

Die Mikroskope, die den Kennbuchstaben M oder P tragen, sind für materialwissenschaftliche, geologische oder mineralogische Untersuchungen vorgesehen.

Alle oben genannten Mikroskope entsprechen der EG-Richtlinie 98/79/EG über In-vitroDiagnostika. Gleichzeitig erfüllen die Geräte die EG-Richtlinien 73/23/EWG betreffend elektrische Betriebsmittel und 89/336/EWG über die elektromagnetische Verträglichkeit für den Einsatz in industrieller Umgebung.

Achtung!

Diese (IVD-) Geräte sind nicht zur Verwendung in der nach DIN VDE 0100-710 definierten Patientenumgebung vorgesehen. Sie sind auch nicht zur Kombination mit Medizingeräten nach der EN 60601-1 vorgesehen. Wird ein Mikroskop mit einem Medizingerät nach EN 60601-1 elektrisch leitend verbunden, so gelten die Anforderungen nach EN 60601-1-1.

Page 89

3. Sicherheitshinweise

3.1 Allgemeine Sicherheitshinweise

Dieses Gerät der Schutzklasse 1 ist gemäß EN 61010-2-101:2002, EN 61010-1:2001, IEC 1010-1:2001, Sicherheitsbestimmungen für elektrische Mess-, Steuer-, Regel- und Laborgeräte gebaut und geprüft.

Um diesen Auslieferungszustand zu erhalten und einen gefahrlosen Betrieb sicherzustellen, muss der Anwender die Hinweise und Warnvermerke beachten, die in dieser Bedienungsanleitung enthalten sind.

Achtung!

3. Sicherheitshinweise

Achtung!

Die in der Bedienungsanleitung beschriebenen Geräte bzw. Zubehörkomponenten sind hinsichtlich Sicherheit oder möglicher Gefahren überprüft worden. Bei jedem Eingriff in das Gerät, bei Modifikationen oder der Kombination mit Nicht-Leica-Komponenten, die über den Umfang dieser Anleitung hinausgehen, muss die zuständige Leica-Vertretung oder das Stammwerk in Wetzlar konsultiert werden!

Bei einem nicht autorisierten Eingriff in das Gerät oder bei nicht bestimmungsgemäßem Gebrauch erlischt jeglicher Gewährleistungsanspruch, sowie die Produkthaftung!

Page 90

3. Sicherheitshinweise

3.2 Elektrische Sicherheit

Allgemeine technische Daten

Elektronikbox Leica CTR5000 (für DM5000 B)

Verwendung nur in Innenräumen. Versorgungsspannung: Frequenz: Leistungsaufnahme: Sicherungen:

90-250 V~ 50-60 Hz max. 290 VA T6,3 A

(IEC 60127-2/3) Umgebungstemperatur: Relative Luftfeuchtigkeit: Überspannungskategorie: Verschmutzungsgrad:

15-35°C

max. 80% bis 30°C

Mikroskop

Verwendung nur in Innenräumen. Versorgungsspannung: Frequenz:

90-250 V~

50-60 Hz Leistungsaufnahme:

DM4000 DM4500 DM5000

max. 180 VA

max. 290 VA Sicherungen:

DM4000 DM4500

DM5000 Umgebungstemperatur: Relative Luftfeuchtigkeit: Überspannungskategorie: Verschmutzungsgrad:

T6,3 A (IEC 60127-2/3) T6,3 A (IEC 60127-2/3) Siehe CTR5000 15-35°C max. 80% bis 30°C II 2

Vorschaltgerät ebq 100*

Verwendung nur in Innenräumen. Versorgungsspannung: Frequenz: Leistungsaufnahme: Sicherungen: Umgebungstemperatur: Relative Luftfeuchtigkeit: Überspannungskategorie: Verschmutzungsgrad:

90-250 V~ 50-60 Hz max. 155 VA 2xT2A (IEC 127) 10-36°C max. 80% bis 30°C II 2

(Siehe beiliegende Anleitung)

Achtung!

Der Netzstecker darf nur in eine Steckdose mit Schutzkontakt eingeführt werden.

Die Schutzwirkung darf nicht durch eine Verlängerungsleitung ohne Schutzleiter aufgehoben werden. Jegliche Unterbrechung des Schutzleiters innerhalb oder außerhalb des Gerätes oder Lösen des Schutzleiteranschlusses kann dazu führen, dass das Gerät gefahrbringend wird. Absichtliche Unterbrechung ist nicht zulässig!

Achtung!

Durch Anschluss an die Erdung können an das Mikroskop angeschlossene Zusatzgeräte mit eigener und/oder extra Netzversorgung auf gleiches Schutzleiter-potenzial gebracht werden. Bei Netzen ohne Schutzleiter ist der Leica-Service zu fragen.

Page 91

Achtung!

Es ist sicherzustellen, dass nur Sicherungen vom angegebenen Typ und der angegebenen Nennstromstärke als Ersatz verwendet werden. Die Überbrückung des Sicherungshalters ist unzulässig.

3. Sicherheitshinweise

3.3 Entsorgung

Nach dem Ende der Produktlebenszeit kontaktieren Sie bitte bezüglich der Entsorgung den Leica Service oder den Leica Vertrieb.

Beachten Sie bitte die nationalen Gesetze und Verordnungen, die z.B. die EU-Richtlinie WEEE umsetzen und deren Einhaltung sicherstellen.

Achtung!

Die elektrischen Zubehörkomponenten des Mikroskops sind nicht gegen Wassereintritt geschützt. Wassereintritt kann zu einem Stromschlag führen.

Achtung!

Schützen Sie das Mikroskop vor zu hohen Temperaturschwankungen. Es kann zur Kondensatbildung und Beschädigung elektrischer und optischer Komponenten kommen. Betriebstemperatur: 15-35°C.

Achtung!

Schalten Sie vor dem Austausch der Sicherungen oder der Lampen unbedingt den Netzschalter aus und entfernen Sie das Netzkabel.

Hinweis!

Wie alle elektronischen Geräte dürfen das Mikroskop, seine Zubehörkomponenten und das Verbrauchsmaterial nicht im allgemeinen Hausmüll entsorgt werden!

Page 92

4. Geräteübersicht

Spezifikation

Kontrastverfahren

Durchlichtachse

Auflichtachse

Leica DM4000 B

Leica DM5000 B

• Durchlicht: DM4000 B: BF, DF, PH, Pol DM5000 B: auch ICT (mot.)

• Auflicht: Fluo

• automatischer Beleuchtungsmanager (mot.. Apertur- und Feldblende, mot. Intensitätsregelung)

• automatische farbneutrale Helligkeitsregelung

• motorischer Shutter

• im Stativ integriert

• mot. 5-fach Filterrevolverscheibe (DM5000 B 8-fach optional)

• mit FIM (Fluoreszenz Intensitätsmanager) zur Reduktion der Lichtintensität in 5 Stufen

• mechanische Booster-Linse zur Verstärkung der Fluoreszenzintensität

• motorischer Shutter

Leica DM4000 M

Leica DM4500 P

• Durchlicht: DM4000 M: BF, DF, PH, ICT,

Pol

DM4500 P: BF, DF, PH, ICT,

Pol (Konoskopie)

• Auflicht: BF, DF, ICR, Pol, Fluo

• im Stativ integriert

• mot. 4-fach Filterrevolverscheibe

• automatischer Beleuchtungsmanager

• DM4000 M: motorischer Shutter

Z-Trieb

Objektivrevolver

• manuell

• manuell, absolut codiert

• DM4000 B:6-fach/7-fach mit M25-Gewinde

DM5000 B: 7-fach (M25) DM5000 B: Mit Objektiv-Prismen-Scheibe (3 Positionen)

• manuell, absolut codiert

• DM4000 M: 6-fach mit M32Gewinde DM4500 P: 6-fach mit M25Gewinde, zentrierbar, codiert

• Aufnahme für DIC Prismen und Pol Kompensatoren (für DM4000 M: optional)

Page 93

4. Geräteübersicht

Spezifikation

XY Tisch

Tubus

Kondensor

Vergrößerungswechsler

(optional)

Bedienelemente

Leica DM4000 B

Leica DM5000 B

• manuell

• Tisch wechselbar

• Länge Koaxialtrieb: 155 mm

• manuell oder motorisiert (DM4500P: manuell)

• optional mit ein oder zwei Kameraausgängen

• DM4500 P: Konoskopie-Modul (Tubusoptik HC P1x/1.6x mit Bertrandlinse, kodiert)

• Kondensorkopf motorisiert

• Kondensorscheibe für Lichtringe, DF-Stop, DIC-Prismen

• automatische Köhlersche Beleuchtung

• optional Polarisator integriert und motorisiert

• manuell

• 3-fach absolut codiert

• 1x; 1,25x; 1,6x

• Bedientasten am Stativ für alle motorisierten Mikroskop-Funktionen

• zusätzlich variable Funktionstasten

• Handräder zum Fokussieren

• LC-Display

• DM5000 B mit Leica SmartTouch

Leica DM4000 M

Leica DM4500 P

• manuell

• DM4000 M:

• Tisch wechselbar

• Länge Koaxialtrieb: 140 mm DM4500 P:

• Pol-Tisch wechselbar

• manuell

• 3-fach absolut codiert

• 1x; 1,5x; 2x

Computer Interface

Software tools

• USB2.0

• Leica Application Suite (LAS)

für Windows

2000, XP, Vista

• mit Plug-ins für:

• Mikroskop- und Kamera-Konfiguration

• Mikroskop- und Kamera-Steuerung

• Image-Acquisition

Page 94

4. Geräteübersicht

Spezifikation

Elektronik-Box Leica CTR5000

Leica DM4000 B

Leica DM5000 B

Nur für Leica DM5000 B: separate Bedieneinheit mit Spannungsversorgung für 100W Halogenlampe. Siehe → S.10 (elektrische Sicherheit)

Leica DM4000 M

Leica DM4500 P

Page 95

4. Geräteübersicht

5 6

Abb. 1 Linke Stativseite mit Advanced Ergotubus AET22 1 Okular

2 Okularstutzen 3 Tubus 4 Objektivrevolver mit Objektiven 5 Präparatetisch mit Präparatehalter 6 Kondensor 7 LC-Display

891011

8 Bedientasten Leuchtfeldblende 9 Umschaltung Durchlicht/Auflicht 10 Bedientasten Aperturblende 11 Bedientasten für Helligkeitseinstellung 12 Fokushandrad mit Grob- und Feintrieb 13 Variable Funktionstasten (werkseitig vorbelegt) 14 Lampen-Justierfenster

Page 96

4. Geräteübersicht

Abb. 2 Rechte Stativseite mit Advanced Ergotubus AET22 15 Lampenhaus für Auflicht

16 Lampenhaus für Durchlicht 17 Durchlichtfilter, optional 18 Durchlichtfilter, optional 19 Variable Funktionstasten (werkseitig vorbelegt) 20 x/y-Koaxialtrieb, höhenverstellbar 21 Handrad für Feinfokus 22 Motorisierter Filterblockwechsler

21 20 19 18 17

Page 97

5. Auspacken

Die Lieferung erfolgt in zwei Packstücken.

Der Stativkarton enthält die folgenden Komponenten:

• Stativ mit integrierter Auflichtachse und Objektivrevolver

• Präparatetisch mit Tischwinkel

• Netzkabel und PC-Verbindungskabel

• CD mit dem Softwarepaket Leica Application Suite (LAS)

• Anleitungen und Liste der Mikroskopvoreinstellung

Der Systemkarton enthält das mikroskopische Zubehör:

• Tubus

• Okulare

Das externe Vorschaltgerät* ebq 100 wird in einer gesonderten Verpackung geliefert.

Für das Mikroskop Leica DM5000 B: Die Elektronikbox Leica CTR5000 wird in einer gesonderten Verpackung geliefert.

Entnehmen Sie zunächst vorsichtig alle Komponenten dem Transport- und Verpackungsmaterial.

Hinweis:

Das Berühren der Linsenoberfläche der Objektive ist möglichst zu vermeiden. Entstehen dennoch Fingerabdrücke auf den Glasflächen, so sind diese mit einem weichen Leder- oder Leinenlappen zu entfernen. Schon geringe Spuren von Fingerschweiß können die Oberflächen optischer Geräte in kurzer Zeit angreifen. Weitere Hinweise im Kapitel “Pflege des Mikroskops”

S. 77.

→

• Objektive

• Kondensor

• Lampenhäuser mit Zubehör

• Montagewerkzeug

• je nach Ausrüstung weiteres mikroskopisches Zubehör wie Filterwürfel, etc.

Achtung!

Mikroskop und Peripheriegeräte auf keinen Fall bereits jetzt an die Steckdose anschließen!

Page 98

5. Auspacken

Aufstellungsort

Das Arbeiten mit dem Mikroskop sollte in einem staubfreien Raum erfolgen, der frei von Öl- und anderen chemischen Dämpfen und extremer Luftfeuchtigkeit ist. Am Arbeitsplatz sollen außerdem große Temperaturschwankungen, direkt einfallendes Sonnenlicht und Erschütterungen vermieden werden. Hierdurch können Messungen bzw. mikrografische Aufnahmen gestört werden.

Zulässige Umgebungsbedingungen: Temperatur 15–35°C Relative Luftfeuchtigkeit max. 80% bis 30°C

Mikroskope in warmen und feucht-warmen Klimazonen brauchen besondere Pflege, um einer Fungusbildung vorzubeugen. Weitere Hinweise in den Kapiteln „Pflege des Mikroskops“ →

S. 77.

Achtung!

Transport

Für den Versand oder Transport des Mikroskops und seiner Zubehörkomponenten sollte die Originalverpackung verwendet werden.

Um Beschädigungen durch Erschütterungen zu vermeiden, sollten vorsorglich folgende Komponenten demontiert und gesondert verpackt werden:

• Schrauben Sie die Objektive heraus.

• Entfernen Sie den Kondensor.

• Entfernen Sie den Objekttisch.

• Nehmen Sie die Lampenhäuser ab.

• Demontieren Sie den Brenner im Lampenhaus 106 z.

• Entfernen Sie alle beweglichen bzw. losen Teile.

Elektrische Komponenten müssen mindestens 10 cm von der Wand und von brennbaren Gegenständen entfernt aufgestellt werden.

Page 99

6. Montage des Mikroskops

6. Montage

Die Mikroskopkomponenten werden sinnvollerweise in dieser Reihenfolge montiert:

• Objekttisch

• Kondensor mit Kondensorkopf

• Tubus

• Okulare

• Objektive

• Lampenhäuser mit Lichtquellen

• Bestückung der Auflicht-Revolverscheibe*

Für die Montage sind nur wenige, universell verwendbare Schraubendreher und Schlüssel notwendig, die im Lieferumfang enthalten sind.

Bei Verwendung von Zwischensystemen und optischem Zubehör kann die Reihenfolge abweichen. Lesen Sie dazu das Kapitel „6.10 Optionales Zubehör“ →

S. 33.

6.1 Objekttisch

Achtung:

Vor dem Montieren des Objekttisches dürfen noch keine Objektive eingeschraubt sein!

• Setzen Sie den Präparatehalter auf den Tisch auf und befestigen Sie ihn mit den beiden Schrauben (3.1).

• Drehen Sie den Kondensorhalter mittels der Kondensorhöhenverstellung (3.2) ganz nach oben, das heißt möglichst nahe an den Tisch.

• Lockern Sie die Tischklemmung (3.3) leicht.

Abb. 3 Mechanischer Objekttisch 1 Fixierschrauben für Präparatehalter 2 Kondensorhöhenverstellung 3 Tischklemmung

Page 100

6. Montage

• Setzen Sie den Tisch von oben an die Schwalbenschwanzführung (4.2) an und schieben Sie den Tisch soweit nach unten, bis das obere Ende der Schwalbenschwanzführung bündig mit dem oberen Ende der Tischklemmung abschließt.

• Ziehen Sie die Tischklemmung (4.1) fest an.

•

Nur für DM4500 P:

Pol-Objektführer*

Objektführer so verstellen, dass die Befestigungsschraube unterhalb der Bohrung (4a.1) sichtbar wird. Objektführer in die Führungsbohrungen des Drehtisches einsetzen und Befestigungsschraube mittels Sechskantschraubendreher anziehen.

Hinweis:

Bei dickeren Präparaten (Leica DM4000 M) kann der Tisch entsprechend niedriger angesetzt werden.

Abb. 4 Ansetzen des Tisches 1 Tischklemmung 2 Schwalbenschwanzführung

Ansetzbarer Objektführer*

Der Objektführer kann links, rechts oder frontal angesetzt werden (ohne Abb.); die Befestigung erfolgt mittels der beiden Klemmschrauben.

Abb. 4a Pol-Drehtisch* und Objektführer Pol 3* 1 Bohrung für Befestigungsschraube 2 Ein-/ausschwenkbarer Hebel für die Halterung

von Objektträgern unterschiedlicher Formate

3 Aufbewahrung für Zentrierschlüssel 4 Rastknopfpaare 5 45°-Rastung 6 Klemmung Tischdrehung

Leica DM5000B, DM4000M User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual