Olympus U-LH100HGAPO, U-LH100HG, U-25ND6-2, U-25ND25-2, U-25ND50-2 User manual

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BX-URA2

BX-RFA U-LH100HGAPO U-LH100HG Power Supply Unit U-25ND6-2 U-25ND25-2 U-25ND50-2 U-RSL6 U-RSL6EM BX-RFSS U-EXBABG U-EXBAUB U-EXBAUG

INSTRUCTIONS

REFLECTED

FLUORESCENCE

SYSTEM

This instruction manual is for the Olympus Reflected Fluorescence System. To ensure the safety, obtain optimum performance and to familiarize yourself fully with the use of this system, we recommend that you study this manual thoroughly before operating the microscope. Retain this instruction manual in an easily accessible place near the work desk for future reference.

This publication is printed on 100% recycled paper

 

A X 7 6 2 5

 

 

CONTENTS

Correct assembly and adjustments are critical for the reflected fluorescence system to exhibit its full performance. If you are going to assemble the reflected fluorescence system yourself, please carefully read section 9, “ASSEMBLY” (pages 30 to 35).

IMPORTANT — Be sure to read this section for safe use of the equipment. — 1-3

I. REFLECTED FLUORESCENCE OBSERVATION

1NOMENCLATURE

2 REFLECTED FLUORESCENCE OBSERVATION PROCEDURE

3USING THE CONTROLS

4-5

6-7

8-15

 

1

General Precautions for Observation.......................................................................................................................

8

 

2

Selecting the Fluorescence Mirror Unit.......................................................................................................

8-10

 

3

Objectives for Various Observation Modes........................................................................................

10-11

 

4

Turning the Power Supply Unit On.............................................................................................................................

11

 

5

Centering the Field Iris Diaphragm..........................................................................................................................

12

 

6

Centering the Aperture Iris Diaphragm ..............................................................................................................

13

 

7

Centering the Mercury Burner ................................................................................................................................

14-15

 

8

Mounting the ND Filters ............................................................................................................................................................

15

 

 

 

4

SIMULTANEOUS FLUORESCENCE OBSERVATIONS

16

 

 

 

 

 

1

Simultaneous Reflected Fluorescence and Phase Contrast Observations..............

16

 

2

Simultaneous Reflected Fluorescence and Transmitted Light Nomarski

16

 

 

Differential Interference Contrast (DIC) Observations ..................................................................

5TROUBLESHOOTING GUIDE

6 SPECTRAL CHARACTERISTICS OF FILTERS

7SPECIFICATIONS

17

18-22

23

8 OPTIONAL MODULES

24-29

1

6-Position Filter Slider U-RSL6...........................................................................................................................

24-25

2

6-Position Barrier Filter Slider U-RSL6EM ..................................................................................................

26

3

Rectangle Field Stop BX-RFSS (for exclusive use with the BX-RFA)....................

27

4

Exciter Balancers U-EXBABG/EXBAUB/EXBAUG (for exclusive use with the BX-RFA) ......

28-29

9ASSEMBLY — See this section for the replacement of the light bulb. — 30-35

9-1

Assembly Diagram ...............................................................................................................................

............................................. 30

9-2

Detailed Assembly Procedures .............................................................................................................................

31-35

II. REFLECTED OBSERVATIONS (BX-URA2 Only)

1

CONFIGURATION OF REFLECTED OBSERVATION SYSTEM

36

 

 

 

 

 

 

2

ASSEMBLY

37

 

 

 

 

 

 

3

FIELD IRIS AND APERTURE IRIS DIAPHRAGM ADJUSTMENTS

37-38

 

 

 

 

 

 

4

OBSERVATIONS

39-42

 

 

 

 

 

4-1

Reflected Light Brightfield/Darkfield Observations ...........................................................................

39

 

4-2

Reflected Light Nomarski Differential Interference Contrast (DIC) Observation ......

40-42

 

4-3

Reflected Light Simple Polarized Light Observation.......................................................................

42

5OPTICAL CHARACTERISTICS «UIS2 (UIS) Series for Reflected Light Observation» 43-44

6 TROUBLESHOOTING GUIDE

45

 

 

■ PROPER SELECTION OF THE POWER SUPPLY CORD .........................................................................

46-47

IMPORTANT

This system employs a UIS2/UIS (Universal Infinity System) optical design, and should be used only with UIS2/UIS microscopes, eyepieces, objectives and condensers for the BX2 series. (Some of the modules designed for the BX series and objectives/eyepieces for the UIS series are also usable. For details, please consult Olympus or the catalogues.) Less than optimum performance may result if inappropriate accessories are used.

The use of a universal reflected fluorescence illuminator has enabled the installation of necessary fluorescence mirror units. By combining the microscopy techniques as shown below, this system can efficiently be used to find fluorescence emission in any area of cells:

1.Reflected fluorescence observation + Transmitted light phase contrast observation

2.Reflected fluorescence observation + Transmitted Nomarski Differential Interference Contrast (DIC) observation

3.Reflected fluorescence observation + Transmitted Light Observation

In addition, the following observations are also by installing a general reflected light observation unit (BX-URA2 only):

1.Reflected brightfield/darkfield observations

2.Reflected Nomarski DIC observation

3.Reflected simplified polarized light observation

This manual describes the instructions for I. Reflected Fluorescence Observations in the first half and those for II. Reflected Light Observations in the second half.

Please find the pages giving you the appropriate instructions for your observation.

SAFETY PRECAUTIONS

1.This system is composed of precision instruments. Handle it with care and avoid subjecting it to sudden or severe impact.

2.The ultrahigh-pressure mercury burner used should be the USH-103OL DC burner (mfd. by USHIO, Inc.) or the HBO103W/ 2 burner (mfd. by OSRAM) that Olympus supplies.

3.Make sure that a mercury burner is attached and that cables are plugged in firmly.

4.The inside of the lamp housing is very hot and hazardous during lighting and for about 10 minutes after turning off. Do not open the lamp housing in this period. (Page 11)

5.Do not apply excessive force to the stoppers which are provided for some functions. Otherwise, the stopper or equipment may be damaged.

6.Do not attempt to open or disassemble the power supply unit because it includes high voltage parts inside.

7.Always use the power cord provided by Olympus. If no power cord is provided, please select the proper power cord by referring to the section “PROPER SELECTION OF THE POWER SUPPLY CORD” at the end of this instruction manual. If the proper power cord is not used, product safety and performance cannot be guaranteed.

Before plugging the power cord to the power outlet, make sure that the main switch of the power supply unit is set to “” (OFF).

8.To ensure safety, be sure to ground the power supply unit. Otherwise, Olympus can no longer warrant the electrical safety performance of the system.

9.Before opening the lamp housing for replacement of the burner or any other internal part, set the main switch to “” (OFF), then unplug the lamp housing connection cable from the power supply unit, and wait for more than 10 minutes

until the lamp housing cools down.

10.The top panel of the lamp housing becomes very hot during operation. To prevent fire hazard, do not block the ventilation through the top panel.

1

Safety Symbols

The following symbols are found on the microscope. Study the meaning of the symbols and always use the equipment in the safest possible manner.

Symbol

Explanation

 

 

Indicates the presence of high voltage (1 kV or more). Take caution to guard against electric shock.

Indicates that the surface becomes hot, and should not be touched with bare hands.

Before use, carefully read the instruction manual. Improper use could result in personal injury to the user and/or damage to the equipment.

lIndicates that the main switch is ON.

Indicates that the main switch is OFF.

Warning indications

Warning indications are placed at parts where special precaution is required when handling and using the System. Always heed the warnings.

Warning indication

· Mercury burner lamp housing

[Warning against

position:

(U-LH100HG, U-LH100HGAPO

high temperature]

 

 

 

· Power supply unit for 100 W

 

 

 

 

 

 

 

mercury burner

[Warning against

 

· ND filters

 

high voltage]

 

(U-25ND6, U-25ND25, U-25ND50)

 

 

 

 

 

 

 

 

 

1Getting Ready

1.This manual pertains only to the reflected fluorescence system. Before using this system together with the BX2 microscope and associated options, make sure that you have carefully read and understood their manuals, and understand how the system should be operated together.

2.The reflected fluorescence system is composed of precision instruments. Handle it with care and avoid subjecting it to sudden or severe impact.

3.Do not use the system where it is subjected to direct sunlight, high temperature and humidity, dust or vibrations.

4.To allow heat from the unit to dissipate well, reserve a distance of at least 10 cm between the lamp housing and power supply unit.

5.The power cord can also be used to cut the power supply in case of emergency. To make this possible, the power supply unit should be installed so that the power cord connector (on the rear of the power supply unit) or the power outlet is easily accessible for unplugging in case of emergency.

2

2Maintenance and Storage

1.To clean the lenses and other glass components, simply blow dirty away using a commercially available blower and wipe gently using a piece of cleaning paper (or clean gauze).

If a lens is stained with fingerprints or oil smudges, wipe it gauze slightly moistened with commercially available absolute alcohol.

!Since the absolute alcohol is highly flammable, it must be handled carefully.

Be sure to keep it away from open flames or potential sources of electrical sparks --- for example, electrical equipment that is being switched on or off.

Also remember to always use it only in a well-ventilated room.

2.With any part of the system other than glass components gets dirty, do not use organic solvents but wipe it with a clean cloth. If the part is extremely dirty, use a lint-free, soft cloth slightly moistened with a diluted neutral detergent.

3.Do not disassemble any part of the system. This could result in malfunctions or reduced performance.

4.The mercury burner has a service life period of 300 hours (USH-103OL, HBO103W/2). When the hour counter on the power supply unit indicates this value, set the main switch to “ ” (OFF) and wait for more than 10 minutes before replacing the mercury burner (Page 33). Unlike electric bulbs, the mercury burner seals high-pressure gas inside. If it

continues to be used after the service life has expired, the glass tube may eventually explode due to accumulated distortion.

5.When not using the microscope, be sure set the main switch to “” (OFF). After confirming that the lamp housing has cooled down sufficiently, cover the microscope with the dust cover for storage.

6.When disposing of the microscope, check the regulations and rules of your local government and be sure to observe them.

3Caution

If the system is used in a manner not specified by this manual, the safety of the user may be imperiled. In addition, the system equipment may also be damaged. Always use the system as outlined in this instruction manual.

The following symbols are used to set off text in this instruction manual.

!: Indicates that failure to follow the instructions in the warning could result in bodily harm to the user and/or damage to equipment (including objects in the vicinity of the equipment).

# : Indicates that failure to follow the instructions could result in damage to equipment. } : Indicates commentary (for ease of operation and maintenance).

3

Olympus U-LH100HGAPO, U-LH100HG, U-25ND6-2, U-25ND25-2, U-25ND50-2 User manual

I. REFLECTED FLUORESCENCE OBSERVATION

NOMENCLATURE

Reflected Illuminator

BX-URA2

Fluorescence Illuminator

BX-RFA

Note The diagram shows the BX-RFA. Parts marked * are not provided on the BX-URA2.

Mirror unit turret

Mirror unit inscription pocket (Page 31)

100 W Mercury Apo Lamp Housing U-LH100HGAPO

100 W Mercury Lamp Housing U-LH100HG

Field iris diaphragm centering screws (Page 12)

x2 screws.

Collector lens focusing knob (Page 14)

Burner centering knobs (Page 14)

Mirror focusing screw (Page 15)

On the rear of the lamp housing.

Aperture iris diaphragm centering screws (Page 13)

x2 screws.

Shutter knob (Page 12)

 

 

\: Shutter OUT

Field iris diaphragm knob (Page 12)

Aperture iris diaphragm knob

{: Shutter IN

 

 

(Page 13)

 

6-position filter inlet (Page 24)

 

 

 

Analyzer/6-position barrier filter slider inlet (Page 26)

 

1

3

2

6

5

4

3

ND filter/* exciter balancer inlet (Pages 15 & 28)

* 6-Position filter slider inlet (Page 24)

4

Fluorescence Mirror Units U-MWU2, etc., total 24 models

}Up to six fluorescence mirror units can be mounted on the BX-RFA or BXURA2.

#Each filter unit includes a dichroic mirror, barrier filter and excitation filter that have been combined according to the excitation method. It is basically not recommended to open a fluorescence mirror unit.

}It is recommended that you use the U-MF2 dummy filter unit (which does not contain a filter) when making your original fluorescence unit. (Page 32) Blank indicator sheets provided with the illuminator can be used to write the names of original fluorescence mirror units.

Indicator sheets

Power Supply Unit

(for 100 W mercury burner)

Burner ON LED

 

Hour counter

Input

 

 

receptacle

 

Output

Main switch

connector

 

I : ON

 

: OFF

 

ND Filters

 

Centering Target

U-25ND6-2, U-25ND25-2, U-25ND50-2

 

U-CST

 

 

 

5

REFLECTED FLUORESCENCE OBSERVATION PROCEDURE

}If you need simultaneous observation of reflected fluorescence observation with the phase contrast observation or transmitted light Nomarski Differential Interference Contrast (DIC) observation, please read Chapter 4, “SIMULTANEOUS FLUORESCENCE OBSERVATION”. (Page 16)

(Controls Used)

(Page)

Preparation

·Attach the fluorescence mirror unit and objective matching the observation method. (Pages 8 to 11)

·Center the mercury burner. (Page 14 or 15)

Set the main switch to “ I ” (ON) and wait for the arc to stabilize.

Place the specimen on the stage.

Engage the fluorescence mirror unit matching the specimen in the light path.

Engage the objective in the light path and focus on the specimen.

Engage an ND filter in the light path as required.

Adjust so that the entire field is uniform and brightest.

@ Main switch

(P. 11)

² Specimen holder ³ X-/Y-axis knobs

| Mirror unit turret

ƒ Revolving nosepiece

… Coarse/fine adjustment knobs

† ND filters

(P. 15)

‡ Collector lens focusing knob

(P. 14)

 

 

Adjust the field iris diaphragm.

Š Field iris diaphragm knob

(P. 12)

 

 

 

 

Adjust the aperture iris diaphragm.

‰ Aperture iris diaphragm knob

(P. 13)

 

 

 

 

 

 

 

Start observation.

 

 

}Engage the shutter if you interrupt observation for a short time.

‹ Shutter knob

(P. 12)

6

† ‡

|

ƒ

Š

³

@

²

} Make a photocopy of the observation procedure pages and post it near your microscope.

7

USING THE CONTROLS

1General Precautions for Observation

1.Verify that the power supply voltage and frequency match the requirements inscribed on the Rating plate.

2.Make sure that the power cord and connecting cables are plugged in securely.

3.If you perform only transmitted light phase contrast or transmitted light DIC observations, leave one cube position on the turret empty. This allows for transmission of white light.

The turret must always be set to one of the click position. If it is deviated from a click position, the cover may be deformed by heat.

4.Enlarge the field iris diaphragm so it just circumscribes the field of view. If decentered, center it using the Allen screwdriver.

5.Always use immersion for oil immersion objectives.

6.If you use an objective with correction collar such as the UPlanSApo40X, UPlanFLN60X, UPlanApo40X or PlanApo40X, you can correct variations in cover glass thickness by adjusting the correction collar.

Correction procedure

If the cover glass thickness is known, match the correction collar to the cover glass thickness using the collar scale provided. If the thickness is not known, turn the collection collar and adjust the fine adjustment knob to where the image is as sharp as possible.

7.Engage the shutter if you interrupt observation for a short time.

(Turning the mercury burner ON and OFF repeatedly will significantly shorten the life span of the burner.)

8.Color fading of specimens

This system features high excitation light intensity to ensure bright observation of dark fluorescence specimens.

In consequence, after long period of observations using high-power objectives, the colors of specimens will fade quicker than usual, causing the view (contrast) of fluorescent images to deteriorate.

In such a case, slightly reduce the excitation light intensity to slow color fading down and improve the fluorescence images.

To reduce the excitation light intensity, use ND filters or aperture iris diaphragm as far as the observation is not affected or use the shutter to limit the exposure of specimen to more than necessary light.

Commercially-marketed color fading protection agent (DABCO, etc.) can also delay fading of specimen colors. The use of

fading protection agent is recommended especially when you perform high-magnification observations frequently.

#Remember that the fading protection agents cannot be used with certain kinds of specimens.

2Selecting the Fluorescence Mirror Unit

Select the fluorescence mirror unit which matches the fluorochrome in use.

#Never mount or use the U-MBF3 brightfield mirror unit together with a with a mirror unit for fluorescence. The U- MBF3 brightness is excessive and injury to the eyes could occur. If this type of mirror unit is to be used together with a mirror unit for fluorescence, use the U-MBFL3 mirror unit equipped with a built-in ND filter or add a 3% ND filter to the U-MBF3.

}Use according to the excitation wevelength:

Olympus has prepared some sets of fluorescence mirror unit combined with appropriate filters which are variable depending on wavelengths.

The wide-band (W) set is normally used. There may be cases, however, where superwide-band (SW) or Narrow-band (N) sets are recommendable.

@Extremely weak fluorescence brightness

 

 

Use the super-wide band (SW).

 

 

(B- and G-excitation only):

}With the SWB, strong autofluorescence may reduce

 

 

 

image contrast.

² Specimens emitting strong autofluorescence:

 

Use the narrow band (N).

 

 

 

 

}The fluorescence bright is somewhat reduced.

8

Dichroic Mirror and Filter Configurations of Fluorescence Mirror Units

Excitation

Mirror Unit

Dichroic Mirror

Excitation Filter

Barrier Filter

Fluorochromes

Method

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

U-MWU2

 

BP330-385

 

· Autofluorescence observation

U

 

DM400

 

BA420

· DAPI: DNA staining

U-MNU2

BP360-370

 

 

 

· Hoechest 33258, 33342: Chromosome

 

 

 

 

 

 

 

 

 

 

 

· Catecholamine

V

U-MNV2

DM455

BP400-410

BA455

· Serotonin

 

 

 

 

 

· Tetracyline: Bones, teeth

 

 

 

 

 

 

 

U-MWBV2

 

BP400-440

 

· Quinacrine, quinacrine mustard:

 

 

 

 

 

Chromosome

 

 

 

 

 

BV

U-MNBV2

DM455

BP420-440

BA475

· Thioflavine S: Lymphocyte

 

 

 

· Acriflavine: Nucleic acid

 

 

 

 

 

 

 

 

 

 

· ECFP

 

U-MWB2

 

BP460-490

 

· FITC: Fluorescent antibody

B

 

 

 

 

· Acridine orange: DNA, RNA

U-MNB2

DM500

BP470-490

BA520IF

· Auramine: Tubercle bacillus

 

 

 

 

 

 

U-MSWB2

 

BP420-480

 

 

 

 

· EGFP, S65T, RSGFP

 

 

 

 

 

 

IB

U-MWIB3

DM505

BP460-495

BA510IF

 

 

 

 

U-MNIB3

BP470-495

 

 

 

 

 

 

 

 

 

 

 

 

U-MWG2

 

BP510-550

 

· Rhodamine, TRITC: Florescent antibody

 

 

 

 

 

· Propidium iodide: DNA

G

U-MNG2

DM570

BP530-550

BA590

· RFP

 

 

 

 

 

 

U-MSWG2

 

BP480-550

 

 

 

 

 

 

 

 

 

 

 

IG

U-MWIG3

DM570

BP530-550

BA575IF

 

 

 

 

 

 

 

IY

U-MWIY2

DM600

BP545-580

BA610IF

Texas Red: Fluorescent antibody

 

 

 

 

 

 

Color Separation Filter Combinations

 

 

 

 

 

 

 

 

 

 

 

 

 

 

For observing only the U-excitation stain,

U

U-MNUA2

DM400

BP360-370

BA420-460

when using U-excitation stain together

 

 

 

 

 

with FITC.

 

 

 

 

 

 

 

U-MWIBA3

 

BP460-495

 

For observing only the B-excitation stain,

IB

 

 

when using B-excitation stain with TRITC

 

DM505

 

BA510-550

U-MNIBA3

BP470-495

 

 

 

or Texas Red.

 

 

 

 

 

 

 

U-MWIGA3

 

BP530-550

 

For observing only the G-excitation stain,

G

DM570

BA575-625

when using G-excitation stain together

 

 

U-MNIGA3

BP540-550

 

 

 

with Cy5.

 

 

 

 

 

 

Mirror Unit Name Meaning

U - M N I B A 2

Model number (2 or 3) For color separation

Excitation (U, V, BV, B, IB, G, IG or IY)

Bandwidth (SW: Superwide band. W: Wide band. N: Narrow band.)

Mirror unit

Universal

9

Exclusively for Fluorescent Proteins

Excitation

Mirror Unit

Dichroic Mirror

Excitation Filter

Barrier Filter

Fluorochromes

Method

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

CFP

U-MCFPHQ

DM450HQ

BP425-445HQ

BA460-510HQ

For ECFP

 

 

 

 

 

 

GFP

U-MGFPHQ

DM485HQ

BP460-480HQ

BA495-540HQ

For EGFP

 

 

 

 

 

 

YFP

U-MYFPHQ

DM505HQ

BP490-500HQ

BA515-560HQ

For EYFP

 

 

 

 

 

 

RFP

U-MRFPHQ

DM565HQ

BP535-555HQ

BA570-625HQ

For RFP

 

 

 

 

 

 

Mirror Unit Name Meaning

U - M C F P H Q

High Quality

CFP/GFP/YFP/RFP

Mirror unit

Universal

3Objectives for Various Observation Modes

UIS2 Series

Objective

 

Reflected light

Phase contrast

Transmitted light DIC

 

fluorescence

difference

 

 

 

 

UPlanSApo

4X

 

¦

¦

 

10X

 

¦

¦

 

20X

 

¦

¦

 

20X O

¦

¦

 

40X

 

¦

¦

 

60X W

¦

¦

 

60X O

¦

¦

 

100X O

¦

¦

 

 

 

 

 

 

PlanApoN

60X

O

¦*

¦

 

 

 

 

 

 

UPlanFLN

4X

 

¦

 

10X

 

¦

¦**

¦

 

20X

 

¦

¦**

¦

 

40X

 

¦

¦**

¦

 

40X O

¦

¦

 

60X

 

¦

 

60X OΙ

¦

¦**

¦

 

100X O2

¦

¦**

¦

 

100X OΙ2

¦

¦

 

 

 

 

 

 

¦ : Recommended combination. ¦* : Slightly inferior in U-excitation.

––: Not usable, or applicable objective is not available.

¦** : A phase contrast (Ph) objective is necessary for phase contrast observation.

10

UIS Series

Objective

 

Reflected light fluorescence

Phase contrast

Transmitted

 

 

 

 

U, V, BV

B, IB, G, IY

difference

light DIC

 

 

 

 

 

 

 

 

 

 

UPlanApo

4X

 

¦

¦

 

10X

 

¦

¦

¦**

¦

 

10X O

 

¦

¦

¦

 

10X W

 

¦

¦

20X

 

 

¦

¦

¦**

¦

 

20X

O3

¦

¦

¦

 

40X

O Ι 3

¦

¦

¦

 

40X

¦

¦

¦**

¦

 

60X

 

¦

¦

 

60X

W3

¦

¦

¦

 

100X

O Ι 3

¦

¦

¦**

¦

PlanApo

40X

 

¦

 

60X

O3

¦

¦

¦**

¦

 

100X

O3

¦

 

 

 

 

 

 

 

UPlanFI

4X

 

¦*

¦*

 

10X

 

¦*

¦*

¦**

¦

 

20X

 

¦*

¦*

¦**

¦

 

40X

O Ι 3

¦*

¦*

¦**

¦

 

60X

¦

¦

¦**

¦

 

100X

O, OΙ3

¦

¦

¦**

¦

 

 

 

 

 

 

 

UApo

20X

3/340

¦

¦

¦

 

20X

W3/340

¦

¦

¦

 

40X

3/340

¦

¦

¦

 

40X

OΙ 3/340

¦

¦

¦

 

40X

W3/340

¦

¦

¦

 

 

 

 

 

 

 

¦: Recommended combination.

¦* : Usable, but image be dark depending on NA.

––: Not usable, or applicable objective is not available.

¦** : A phase contrast (Ph) objective is necessary for phase contrast observation. The Ph objective is not available for the UPlanFI100XOI3.

4Turning the Power Supply Unit On

Set the main switch to “ I ” (ON). The arc will stabilize in 5 to 10 minutes after ignition.

}The discharge type mercury burner may not be ignited from the beginning on rare occasions due to its characteristics. In this case, set the main switch to “ ” (OFF), wait for 5 to 10 seconds, then set it again to “ I ” (ON).

#To extend the mercury burner life, do not turn the mercury burner off for 15 minutes after ignition.

#The mercury burner cannot be reignited until the mercury vapor has cooled down and liquefied. Before re-igniting a mercury burner, wait for about 10 minutes after the last time it was turned off.

}For the shake of safety, the power supply to the lamp housing is shut down if the lamp housing is opened while the burner is on. If this happens, set the main switch to “ ” (OFF), wait for more than 10 minutes, then set it again to “ I ” (ON). Do not open the lamp housing until it has cooled down enough.

#To reset the hour counter, hold its reset button till “000.0” is displayed.

11

²

5 Centering the Field Iris Diaphragm

(Fig. 1)

 

 

³1. Close the light path by sliding the shutter knob @ to position marked {.

 

2.

Engage the B or IB mirror unit in the light path by rotating the turret.

 

 

(If these mirror units are not available, engage another fluorescence mir-

 

 

ror unit in the light path.)

@

3.

Open the light path by sliding the shutter knob to position marked \.

 

4

Engage the 10X objective in the light path, place the specimen on the

 

 

stage and bring the image into approximate focus.

 

5.

Pull out the field iris diaphragm knob ² to minimize the field iris diameter.

 

6.

Fit the Allen wrench provided with the microscope frame in the two field

Fig. 1

iris centering screws ³ and adjust so that the iris image comes at the center of the field of view.

7.While pushing in the field iris diaphragm knob ², enlarge the field iris diaphragm until the field iris image inscribes the field of view. If eccentricity is found after this, try centering again.

8.Enlarge the iris diaphragm until the iris image becomes almost the same size as (i.e. circumscribes) the field of view.

Effects of Field Iris Diaphragm

The field iris diaphragm restricts the diameter of the beam of light entering the objective and thus excludes extraneous light, improving image contrast. The field iris diaphragm also functions to prevent color fading of fluorescent light in other part than the observed region.

To exclude extra light, set the field iris diaphragm knob ² on the fluorescence illuminator according to the objective power, so that the image of the field iris diaphragm just circumscribes the field of view.

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