BHT SYSTEM MICROSCOPE Instruction Manual July 1986

CONTENTS

STANDARD EQUIPMENT . . . II.

NOMENCLATURE

Ill.

ASSEMBLY

IV.

IDENTIFICATION AND FUNCTION OF VARIOUS COMPOhlENTS

V.

OPERATION

...

...

A . Switching on the Light Source

1 voltage Adjustment and Light Intensity I B.

Placement of a Specimen Slide

pizGq... kwecimen Slide 1

C.

Observation Tube... 1. lnterpupillary Distance Adjustment

2. Diopter Adjustment 3. Light Path Selector D. Condenser Adjustment . 1. Condenser Centration Field lris Diaphragm

I

Aperture lris Diaphragm E.

I

Focusing Adjustment... 1. Tension of Coarse Adjustment Knobs and Fine Adjustment l ~ s of e Rubber Cap for Fine Adjustment Knob 2. Pre-Focusing Lever

3. Adjustment of Stage Block Height F.

Use of Immersion Objectives...

G.

Photomicrography

VI.

OPTICAL DATA

VII.

TROUBLESHOOTING . .

/

..

I.

STANDARD EQUIPMENT Model

Component Microscope stand

BHT-F

I Line cord

UYCP Binocular tube

Observation tubes

1

BH2-B 130

1 1 1 1 1 1

I Quintuple revolving nosepiece Square mechanical stage with right-hand low drive coaxial controls

BH2-TR30 BH2-5RE

0

Abbe condenser

BH2-CD

Swing-out condenser

BH2-SC

1

1

0

0

1

6V20WHAL

1

1

2

D Plan 4X. D Plan 10X. D Plan 40X D Plan 100X (oil)

0

WHKIOX

0

1

1 each

Photo eyepiece

1

-

D Ach. 4X. D Ach. IOX, D Ach. 40X, D Ach. 100X (oil)

I Eyepieces

1

1

-

Halogen bulbs

1

1

1

-

LS-20H

Halogen lamp holder

0

0

BH2-SVR

Condensers

r

1

1

o -

Trinocular tube

0 bjectives

1

1

1

1

N F K3.3X

1

2

2

1

2

1

0

0

1

Filter

I Immersion oil, bottled 1 VinyTdust cover

1

1

1

1

1

II.

NOMENCLATURE

The Model BHT consists of various components and interchangeable accessories as shown in the photo below. A wide variety of combinations, standard or optional, is available according t o your requirements.

Observation tube

Microscope stand Revolving

Objective

Stage

Condenser

Base

"I Halogen lamp holder

-

Ill.

-

ASSEMBLY This picture illustrates the sequential procedure o f assembly. The numbers indicate the order o f assembly of various components. Remove dust caps before mounting components. Take care t o keep all glass surfaces clean, and avoid scratching the glass surface.

NOTE: For numbers O @ and 0 please refer t o explanations in detail on the next page.

Eyepiece

Condenser clamping screw

Halogen bulb

Condenser (The N.A. scale engraved on the condenser should face the microscope front.)

Microscope stand Line cord

0 Outlet

Explanations in detail @ Mounting the stage 1) Loosen the stage clamping screw @ by rotating counterclockwise. (Fig. 1) 2) Insert the stage into the mounting dovetail of the microscope stand slowly and lock with clamping screw.

Fig. 1

@ Mounting the revolving nosepiece

a.

1) L-oosen the nosepiece clamping screw (Fig. 2) 2) Aligning the nosepiece dovetail slide t o the mounting block @ , push in the nosepiece slowly all the way.

NOTE: Do not tilt or rock the nosepiece while inserting into the mounting block.

@ Mounting the observation tube 1) Loosen the clamping knob @ fully. Pull spring-loaded clamping knob @ . This will cause the locating pin @ to withdraw. (Fig. 3) I f the pin does not, loosen the screw further until the pin withdraws. 2) With clamping knob @ pulled out, insert the circular dovetail of the observation tube into the ring dovetail. 3) Tighten the clamping knob. Fig. 3

IV.

IDENTIFICATION AND FUNCTION OF VARIOUS COMPONENTS

Light path selector knob

Tension adjustment ring Clockwise rotation increases coarse adjustment tension.

Specimen holder

Stage clamprng screw Loosen the screw and the stage can be rotated

Coarse adjustment knob

/ Aperture ~ r i s diaphragm ring

Coarse adjustment range 2 6 m m

Condenser centering knobs Y-axis low drive control knob Y excursion range 50mm

Fine adjustment knob Graduated in increments

Voltmeter

/ /

Slidinq voltaqe control lever Voltage lowers as the lever is pulled toward the microscope front.

Photo tube

adjustment

Condenser height adjustment knob

Pre-focusing lever

7

Filter mount

A r r o w m a r k 6 +O indicates increase in diaphragm diameter.

L ~ n ecord

Summary of Putting the Microscope into Operation Model BHT Match the voltage selector switch to local mains voltage (page 9). Switch on the light source (page 9). Place a specimen slide on the mechanical stage (page 9). Coarse focus with the 1 OX objective (page 10. 1 3 ) . Make interpupillary distance and diopter adjustments (page 11). Adjust the condenser position (page 12). Swing in the desired objective. Adjust light intensity. Fine focus. Adjust aperture iris diaphragm and field iris diaphragm (page 12).

Adjustment of Illumination System for Various Objective Powers

1

I

Objective

1 magnification

1

Condenser Achromaticcondenser BH2-AAC

Abbe condenser

Swing-out condenser

Low power condenser

BH2-CD

BH2-SC

BH2-UL-C

Compatible

Swing in top lens

*N.A. is somewhat low, but st111compatible with a 100X objective. (Cut off this page at dotted line and put i t on the wall near the microscope for use as a reminder of microscopic procedure.)

OLYMPUS

V.

OPERATION

A.

Switching on the Light Source

1) Ascertain that the voltage selector switch @ is set to conform with the local mains voltage. (Fig. 4) If the switch is not correctly set, adjust i t by means of the Allen wrench provided or a screwdriver.

2) Place the sliding voltage control lever on the right side of the microscope base to a position closest to you (low voltage position). Switch on the light source. (Fig. 4) Voltage Adjustment and Light Intensity

I

Fig. 4

1

As you push the control l e v e r a in the direction of the arrow in order to obtain increasing intensity (Fig. 5), the LED readout @ will display the lamp voltage. B.

Placement of a Specimen Slide

1) Rotate the coarse adjustment knobs in the direction of the arrow to rack down the stage so that a specimen slide can be placed on the stage. (Fig. 6)

Fig. 5

NOTE: The rotation of the coarse and fine adjustment knobs in the direction of the arrow will rack down the stage.

2) Opening the spring-loaded finger of the specimen holder with one hand, place a specimen slide inside the holder. (Fig. 7) When the slide comes in contact with the back of the specimen holder, slowly return the spring-loaded finger.

Fig. 6

WARNING: If the spring-loaded finger is returned quickly, it may cause damage to the specimen slide.

Fig. 7

-

I Cover Glass I An Olympus objective engraved "160/0.17" requires a cover glass of 0.17mm thickness. If the numerical aperture of the objective is 0.7 or higher (except immersion objectives) and no correction collar is provided, the resolving power deteriorates to a great extent if cover glass thickness deviates from the above listed value. IVOTE: In some countries a 0.17mm cover glass corresponds to a designation of #I%. A cover glass (0.4mm thick) for blood counting, etc. can be used with Olympus objectives except D Plan 40X, S Plan Apo 40X and S Plan 100X.

1 Specimen Slide ( Specimen slides 0.8 mm to 1.5 mm thick are recommended for Olympus objectives. Specimen slides 0.8 mm to 1.2 mm thick are recommended for the darkfield condenser and the differential interference contrast condenser.

3) Bring the portion of the specimen for observation into the light path by means of the low drive control knobs. (Fig. 8)

* Tighten the stage clamping screw @ in the microscope front.

1 stage 1 The specimen holder can accommodate two standard specimen slides simultaneously. The specimen holder is removable to obtain a large unobstructed stage surface to hold specimens up to 55 mm x 85 rnm. To rotate the stage loosen the stage clamping screw @ and holding this screw, rotate the stage into the desired direciton. (Fig. 9)

Fig. 9

Q Stage clips for use with immersion objectives. (Fig. 10) A pair of stage clips are optionally available to hold the specimen on the stage, eliminating a specimen drag caused by immersion oil between slide and stage surface. The clips can be inserted into the holes @ provided on the specimen holder.

Fig. 10

Observation Tube lnterpupillary Distance Adjustment

1 ) Click the 10X objective into position. 2) Looking through the eyepieces with both eyes, adjust the interpupillary distance of the binocular tube by adjusting the knurled dovetail slides @ of the right and left eyepiece tubes with both hands until perfect binocular vision is obtained. (Fig. 11) Diopter Adjustment

1) Look at the image through the right eyepiece with your right eye and focus on the specimen with the fine adjustment knobs. 2) Next, look at the image through the left eyepiece with your left eye and rotate the diopter adjustment ring @ t o focus on the specimen without using the coarse and fine adjustment knobs. (Fig. 12)

Fig. 12

Light Path Selection 1 ) The trinocular tube is provided with a light path selector knob t o direct the light to the observation tube andlor to the photo tube in 3 positions. (Fig. 13)

Fig. 13

I

Knob Position

/

1

Pushed in all the way (V)

Pulled out halfway (C. V.)

100% into binocular tube

20% into binocular tube 8 0 % into ~ h o t tube o

1

Pulled out all the way (C)

I

1

into photo tube

@ Observation Application

@ Dark specimens

Photomicrography (fo-

The indicator plate is provided at the knob port to summarize the usage of the above table; i t can be consulted before operating the knob.

V: Viewer (white letter) CV: Camera & viewer (yellow-green letters) C: Camera (red letter) The colors of the letters correspond with the color bands on the knob shaft.

D. Condenser Adjustment

,

1. Condenser Centration 1) Stop down the field iris diaphragm with knurled ring @ by rotating in the direction of the arrow. (Fig. 14) 2) Use the condenser height adjustment knob @ to move the condenser up and down until an image of the field diaphragm can be seen clearly in the eyepieces. The rotation of the knob in the direction of the arrow lowers the condenser. Field iris diaphragm image Field of view

-7g+('J

Fig. 15

3) Bring the field iris diaphragm image into the center of the field of view with the two condenser centering knobs @. (Fig. 14)

4) Widen the diameter of the iris diaphragm progressively. If the polygonal image of the iris diaphragm becomes inscribed in the field i t means that the field diaphragm i s centered. (Fig. 15) Field Iris Diaphragm The field iris diaphragm controls the diameter of the ray bundle impining on the specimen surface and therefore, by stopping down the field diaphragm until i t is slightly larger than the field of view, i t can reduce stray light, which in turn increases image definition and contrast.

In order to achieve optimum objective performance, the opening of the aperture iris diaphragm should be matched to the numerical aperture of the objective in use. I t is often preferable, however, to stop down the aperture diaphragm slightly more than indicated by the objective N.A. This will result in better image contrast, increased depth of focus and a flatter field. After completing focus adjustment, remove one of the eyepieces from the observation tube and look into the empty eyepiece tube. As you stop down the aperture iris diaphragm, the image of the Opening of the aperture diaphragm iris diaphragm can be seen in the objective pupil. Adjust the opening of the Objective exit pupil diaphragm to match the N.A. of the objective in use. I f the specimen is low in contrast, i t is recommended to stop down Fig. 16 to 70% -- 80% of the objective N.A. (Fig. 16)

I

Focus Adjustment Tension of Coarse Adjustment Knobs and Fine Adjustment.

r r

Although the tension of the coarse adjustment knobs has been already adjusted for optimum performance by the manufacturer, 'i i t is possible t o personally adjust the tension of the coarse adjustment for either heavy or light movement depending on the operator's preference by rotating the tension adjustFig. 17 ment ring @. (Fig. 17) The ring can be rotated by inserting a screwdriver into one of the holes on the periphery of the ring. The clockwise rotation (in the direction of the arrow) tightens the coarse adjustment knobs. Do not loosen the ring too much, because the stage may drop or the fine adjustment knobs may slip.

'3

la

NOTE: Do not rotate the right and left coarse adjustment knobs in the opposite directions simultaneously. If the stage drops and the specimen goes out of focus, the tension adjustment ring is too loose. Tighten the ring.

I Use of Rubber Cap for Fine Adjustment Knob I

-b

Attaching this cap over the fine adjustment knob increases the sensitivity of the fine focusing motion. (The rubber cap is optionally available.)

2. Pre-Focusing Lever

This lever @ is provided to prevent possible contact between specimen and objective as well as to simplify coarse focusing. (Fig. 18) The lever is locked after coarse focus has been accomplished. This prevents further upward travel of the stage by means of the coarse adjustment knobs, and automatically provides a limiting stop if thestage is lowered and then raised again. The pre-focusing lever does not restrict fine focusing.

Fig. 10

3. Adjustment of Stage Block Height In addition to the vertical movement of the stage by means of coarse and fine adjustments, the stage block height can be changed for observation of specimens 'which are thlcker than standard slides, e.g. chambers, flasks, etc. with much larger thickness. The stage block height can be adjusted b y Fig. 19 loosening the stage block locking screw @ with the Allen wrench provided and retightening i t at the upper position. Then, dislocate the lower limit stop pin beneath the stage block into a lower tapped hole. After lowering the stage block, reclamp the stage block locking screw (Fig. 19)

a.

F.

Use of Immersion Objectives

1 ) Focus the specimen with a low power objective.

2) Put a drop of immersion oil on the specimen slide and the front lens o f the immersion objective. 3) Turn the revolving nosepiece t o bring the immersion objective into the light path, and focus with the fine adjustment knobs. NOTE: @ For immersion condensers such as an achromatic-aplanatic condenser or Abbe condenser, remove the specimen from the mechanical stage and place a drop of immersion oil on the front lens of the condenser. Then, place the specimen on the stage and slowly raise the condenser until firm contact with the underside of the specimen slide is made. @Care should be taken t o prevent oil bubbles from forming in the oil film between condenser and specimen slide. If any, re-apply immersion oil, for these bubbles greatly deteriorate the lens performance. @ A f t e r use carefully wipe off the immersion oil deposited on the lens surfaces with gauze moistened with xylene. Never leave oil on the lens surfacesafter use as oil remnants will seriously impair the performance of the lens system.

G.

Photomicrography The Olympus Photomicrographic Equipment Model PM-IOAD is uniquely qualified t o be used with the BHT microscope for routine and advanced photomicrography. A separate, detailed instruction manual is available for the PM-TOAD camera system. For quick reference, however, you may want to refer to the following pointers when using the PM-IOAD.

1. Photographic Eyepiece Use NF K photo eyepieces for photomicrography. Insert the eyepiece into the eyepiece tube o f the photo tube. (Fig. 20)

Fig. 20

2. Mounting the Photographic Unit Slip the body of the photographic unit over the photo tube. Align the dots on photo tube and the PM-1OAD body and clamp the camera unit t o the photo tube. (Fig. 21)

3. Setting the Light Path Selector Refer t o section C.3. on page 11.

Fig. 21

4. Focusing Procedure Use the field of view eyepieces for focusing on the fi Im plane. Each field of view eyepiece has a focusing front lens and a reticle with 4 frames, each frame indicating the area covered by a specific power NFK photo eyepeice. (Fig. 22). The number at each frame indicates the magnification of the photo eyepiece. The ,:f;~l i,mage in the field of view eyepiece and the - image on the film plane are in focus at the & '. JJLll same time. Several type field of view eyepieces are available, according to the film size employed. Fig. 22

i')

Field of view eyepiece

Attachment camera

35WH K10X

PWH KIOX

4X5WHKlOX

MHWHKIOX

35 mm Back

3%" x 4%"

4" x 5" Sheet Film or Polaroid Film Holder

16 mm Bolex camera 120 Roll Film Holder

Polaroid Back

1) Select the field of view eyepiece matching the camera back in use and insert i t into the right eyepiece tube of the trinocular tube, aligning locating groove and locating pin.

2) While looking through the field of view eyepiece, rotate the eyepiece front lens in screw mount to focus on the double cross lines in the field. For sharp focusing with objectives 4X or lower, the focusing magnifier FT is recommended. 3) Bring the specimen detail to be photographed within the frame corresponding to the power of the NFK eyepiece in use and focus on the specimen with the microscope fine adjustment knobs. Make sure the light path selector knob on the observation tube is either on the white (V) or yellow-green (CV) band. 4) I t is recommended to tighten the tension adjustment ring considerably t o prevent the stage from dropping during long exposures.

VI. OPTICAL DATA D Plan Ach.

D Achromat 10X

40X

lOOX*

4X

1OX

40X

10OX1

0.10

0.25

0.65

1.30

0.10

0.25

0.65

1.25

18.2

7.2

0.6

0.20

7.03

7.4

0.27

0.17

Focal length 30.03

16.9

4.58

1.91

34.23

17.5

4.67

1.75

Resolving power (fi)

3.36

1.34

0.52

0.26

3.36

1.34

0.52

0.27

40X

100X

400X

171.6

27.45

3.0

0.7

171.6

27.45

3.0

0.7

W.D. (mm)

Eyepiece

Total mag. WHKlOX Focal (Field depth (PI number . 20) Field of view (mm)

2

* Immersion objectives The resolving power and focal depth are obtained w i t h fully opened aperture diaphragm. Technical terms: Working distance:

The distance from the cover glass t o the nearest point of the objective.

Numerical aperture:

The N.A. represents a performance number which can be compared t o the relative aperture (f-number) of a camera lens. The N.A. values can be used for directly comparing the resolving powers o f all types of objectives. The larger the N.A., the higher resolving power.

Resolving power:

The ability o f a lens t o register small details. The resolving power of a lens is measured b y its ability t o separate t w o points.

Focal depth:

The distance between the upper and lower limits o f sharpness in the image formed by an optical system. As you stop down the aperture iris diaphragm, the focal depth becomes larger. The larger the N.A. o f an objective the shallower the focal depth.

Field number:

A number that represents the diameter i n m m of the image o f the field diaphragm that is formed b y the lens in front o f it.

Field o f view diameter:

The actual size of the field o f view in m m on the object surface.

VII. TROUBLESHOOTING If you are unable t o obtain full performance from your microscope, please consult with the table below as pointers for troubleshooting.

Remedy Cause I Phenomenon I 1. Optical System I I a) With illuminator switched I Field iris diaphragm is not I Open diaphragm to proper on, the field o f view is dark.

b) Field of view is cut o f f or illuminated irregularly.

pulled out to C position.

position.

Light path selector lever is stopped m idway.

Click i t into proper position according to your purpose.

Nosepiece is not clicked into place.

Slightly rotate nosepiece until i t clicks into place.

Nosepiece is not correctly

Insert nosepiece dovetail into

1 mounted. 1

I microscope frame all the way, I then lock.

The power of objective used exceeds the illumination capacity of condenser.

Choose a condenser to meet your purpose.

1 Condenser is not centered. ( Center condenser. Field iris diaphragm is stopped down excessively.

Open diaphragm to proper diameter.

c) Dust or dirt is visible in the field of view.

Remove dust, etc. 'lean front lenses.

1 Dirty specimen. I ~ u sont eyepiece. d) Excessive image contrast.

I

Condenser much.

is

lowered too

Aperture iris diaphragm is stopped down excessively.

I I Adjust condenser height. Open diaphragm to proper diameter.

1

Cause

Remedy

Non Olympus objectives are used.

Use Olympus LB series objectives.

Phenomenon e) Resolution problems: lmage is not sharp. Insufficient contrast. lmage details lack definition.

microscope frame all the way,

positioned in the light path.

I Objective correction collar is I Rotate correction collar, keep1 ing specimen in fine focus

I not adjusted.

until optimum resolution is

I used with immersion oil.

1 Remove bubbles (and reapply oil).

Bubbles in immersion oil. Immersion oil designated by

1 Olympus is not used. Dirty specimens.

1

Use Olympus immersion oil.

1

Clean.

Dust on condenser lens. f ) Field of view is partially

out of focus, or image is partly out of focus.

g) Specimen image is partially out of focus.

h) Field of view becomes only slightly brighter by increasing voltage.

Nosepiece is not correctly

1 mounted.

lnsert nosepiece dovetail into

( microscope frame all the way,

1

1 then lock.

Objective is not correctly positioned in the light path.

Slightly rotate nosepiece until i t clicks in place.

Specimen is not correctly positioned on stage.

Place specimen slide correctly on stage, and place stage clips open it.

Nosepiece mounted.

is not correctly

lnsert nosepiece dovetail into microscope frame all the way, then lock.

Objective is not correctly positioned in the light path.

Slightly rotate nosepiece until i t clicks into place.

Condenser is not centered.

Center condenser.

Condenser is not correctly centered.

Center condenser.

Condenser

Adjust condenser height.

is

lowered too

2. Electric System

I a) Illuminator i s too bright I Line voltage selector switch Match selector switch to

I

(or too dark) even when

I is not matched with local mains voltage

b) Voltage for illuminator cannot be raised.

mains voltage.

1

I

1

Phenomenon C)

Lamp & o f f

and

1

on I

1

Cause Bulb filament burn out.

is likely t o

1

Remedv

/ Replacebulb. I

I

r Loose electric connections. I Check all connections.

I Line voltage selector switch is I Match selector switch to 1 not matched with local mains mains voltage. 1

d) Bulb burns out frequently.

voltage. Bulb is not standard one.

Use standard bulb.

1 3. Coarse and Fine Adjustments a) Coarse adjustment knob is too tight.

Tension adjustment tightened too much.

b) Stage drops or specimen

goes out of focus during observation due to slipping fine adjustment knobs.

I Loosen ring properly.

ring is

User is trying to raise stage above the focusing limit imposed by the engaged prefocusing lever.

Unlock lever.

Tension adjustment too loose.

Tighten ring properly.

ring is

C) Stage cannot be raised to the upper limit.

Pre-focusing lever is engaged in lower than focusing position.

d) Stage cannot be lowered to the lower limit.

Stage is mounted too low.

e) Objective front lens hits specimen before coming into focus.

Specimen is placed on stage upside down.

Unlock lever.

Raise stage mount with Allen

I wrench.

-

Reverse specimen

1 4. Observation Tubes

1

a) l ncomplete binocular vision.

Correct the interpupillary distance.

l nterpupillary distance is not correctly adjusted.

1 Diopter adjustment is incom- I Complete the diopter adjust- 1 1 plete.

I ment.

Right and left eyepieces are not matched. User is unaccustomed to bi-

I nocular vision.

1

1 5. Stage a) l mage easily goes out of focus when you touch the stage.

1

b) Specimen stops midway on the east-west traverse.

1

Use a pair of matched eyepieces. Prior to looking into the bi-

1 nocular observation tube, look I 1 at a far away obiect.

Stage is not correctly locked.

Clamp stage securely.

Specimen is not correctly po-

Adjust specimen position.

I sitioned.

I

1

I