BHS SYSTEM MICROSCOPE Instructions May 1990

This instruction manual has been written for the use of the Olympus System Microscope Model

BHS. It is recommended that you read the manual carefully in order to familiarize yourself fully with the use of the microscope, so that you can obtain optimum performance from it.

IMPORTANT ObSe~ethe following points: Operation 1.

Always handle the microscope with the care i t deserves, and avoid abrupt motions.

2. Avoid the use of the microscope in direct sunlight, high temperature and humidity, dust and vibration.

3.

Only use the tension adjustment ring for altering the tension of the coarse adjustment knobs. (Do not twist the two coarse adjustment knobs in opposite directions simultaneously, as this will cause damage.)

4.

Make it a point of grounding the microscope to prevent electric accidents.

rn Maintenance 1.

Lenses must always be kept clean. Carefully wipe off oil or fingerprints deposited on the lens surfaces with gauze moistened with a small amount of xylene, alcohol or ether.

2.

Do not use organic solutions to wipe the surfaces of various components. Plastic parts, especially, should be cleaned with neutral detergent.

3.

Never disassemble the microscope for repair. Only authorized Olympus service personnel should make repairs.

4. The microscope should be covered with the vinyl dust cover provided and stored in a place free from humidity.

CONTENTS

1.

STANDARD EQUIPMENT...

2

II.

NOMENCLATURE

...

3

Ill.

ASSEMBLY

...

4

IV.

IDENTIFICATION AND FUNCTION OF VARIOUS COMPONENTS...

6

V.

OPERATION

...

9

A. Switching on the Light Source /voltage Adjustment and Light Intensity I

B.

C.

Placement of a Specimen Slide

pzzzizq... /specimen slide 1

10

...

11

Observation Tube

1. lnterpupillary Distance Adjustment

2. Diopter Adjustment 3. Light Path Selector 0 . Condenser Adjustment... 1. Condenser Centration

12

l ~ i e l dlris ~ i a ~ h r a g m l l ~ p e r t u r elris Dia~hranm1 E.

Focusing Adjustment

...

13

1. Tension of Coarse Adjustment Knobs and Fine Adjustment /use of Rubber C ~ for D Fine Adiustment ~ n o b l

2. Pre-Focusing Lever 3. Adjustment of Stage Block Height F.

Use of Immersion Objectives...

G.

Photomicrography

14

...

16

...

17

VI.

OPTICAL DATA

VII.

TROUBLESHOOTING GUIDE

I.

STANDARD EQUIPMENT Model Component BHS-112 BHS-113 BHS-312 BHS-313 Microscope stand Binocular tube

BHS-F

1

1

1

1

BH2-B 130

1

1

0

0

0

0

1

1

0bservation tubes Trinocular tube

BH2-TR30

Quintuple

BH2-5RE

1

0

1

0

Sextuple

BH2-6RE

0

1

0

1

Square mechanical stage with right-hand low drive coaxial controls

BH2-SVR

1

1

1

1

BH2-SC

1

1

1

1

0

1

0

1

Revolving nosepieces

Swing-out condenser Condensers

Achromatic/ Aplanatic BH2-AAC condenser

Halogen lamp housing

BHS-LSH

1

1

1

1

Halogen bulbs

12V100WHAL-L

2

2

2

2

D Plan 4X, D Plan 1OX, D Plan 20X, D Plan 40X. D Plan 1OOX (oil)

leach

0

1 each

0

S Plan 4X. S Plan 10X. S Plan 20X, S Plan 40X, S Plan 100X (oil)

0

1 each

0

1each

Objectives

Eyepieces

WH K1OX

2

2

2

2

Photo eyepiece

NF K3.3X

0

0

1

1

Line cord

UYCP

1

1

1

1

Filter

KB-4

1

1

1

1

Immersion oil, bottled

1

1

1

1

Vinyl dust cover

1

1

1

1

Allen wrench

1

1

1

1.

II.

NOMENCLATURE

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

Observation tube

7 Microscope stand For biological use.

Halogen lamp housina

Ill.

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

NOTE: For numbers 0 ,0 ,0 and 0 please refer to explanations in detail on the next page.

Circular dovetail mount

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

Halogen bulb

Halogen lamp

Line cord

@

Microscope stand Outlet

Explanations in detail Q Mounting the halogen bulb

1) Releasing the bulb clamping leve$@ of the collector lens @ in the dir tion of of the the arrow, insert two contact halogen bulb into the socket Q. (Fig. 1) (Recommended to use a glove or gauze to handle the halogen bulb.) 2) Secure the bulb in position with the two levers.

?.E

* Before use, wipe off any fingerprints or stains on the bulb.

Fig. 1

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

Fig. 2

0 Mounting the revolving nosepiece 1) Loosen the nosepiece clamping screw @. (Fig. 3) 2) Aligning the nosepiece dovetail sl~deto the mounting block @, push in the nosepiece slowly all the way.

Fig. 3

@ Mounting the observation tube

W'-*"

1) Loosen the clamping knob @ fully. Pull spring-loaded clamping knob @ . This will cause the locating pin @ to withdraw. (Fig. 4) If 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. 4

IV.

IDENTIFICATION AND FUNCTION OF VARIOUS COMPONENTS

Light path selector knob

/

The knob can be operated in 3 positions to deflect the light as desired.

Tension adjustment ring Clockwise rotation increases coarse adjustment tension.

adjustment knob Coarse adjustment range: 2 6 m m

X-axis low drive control knob

Accepts the ND filter holder.

X excursion range: 7 6 mm.

Filter diameter 45mm.

Sliding voltage control lever Voltage lowers as the lever is pulled toward the microscope front.

+ D o not disengage the ND filter for about a few minutes after the use, in order to give time to cool. Other filters (e.g. KB-4 provided, etc.) should be inserted into the filter mount at the light exit on the microscope base.

Photo tube

-. Diopter adjustment ring

Condenser height ad iustment knob

Arrow mark Q + 0 indicates increase in diaphragm diameter.

*This breaker protrudes i t s central part t o cut off the electric power in case of the dimmer circuit trouble (due to short circuit, etc.) or over current. To restore the breaker, press the central part. If the breaker is actuated again, disconnect the line cord from the AC outlet and contact the Olympus Service Center.

Summary of Putting the Microscope into Operation Model BHS A.

Switch on the light source (page 9).

8.

Place a specimen slide on the mechanical stage (page 9).

C.

Coarse focus with the 10X objective.

D.

Make interpupillary distance and diopter adjustments (page 11).

E.

Adjust the condenser position (page 12).

F.

Swing in the desired objective.

G.

Adjust light intensity.

H.

Fine focus.

I.

Adjust aperture iris diaphragm and field iris diaphragm (page 12).

4-

3

0

Condenser Objective

Achromaticaplanatic condenser

magnification

,

I

40X

I

Abbe condenser

.

Compatible

I

Swing-out condenser

Low power condenser

Swing out top lens*

Compatible

1 s w ~ n g~ntop I___

I,$?>.,,:;;?

..

.):.: t.-,,

*To obtain better illumination for photomicrography with a 2X objective, it is recommended to use the ultra-low condenser BH2-ULC, optionally available. **N.A. is somewhat low, but still compatible with a 100X objective. (Cut off this page at dotted line and put it on the wall near the microscope for use as a reminder of microscopic procedure.)

V.

OPERATION

A.

Switching on the Light Source

1) 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. 5)

Fig. 5

[ Voltage Adjustment and Light Intensity I As you push the control levera in the direction of the arrow in order to obtain increasing intensity (Fig. 6), the LED readout @ will display the lamp voltage.

* Two LEDs on the left side indicate the

voltage from OV to 6V, and twelve LEOSon the right side from 6.5V t o 12V in 0.5V increments. The indication with the letters "PHOTO" can be used as a guide line for color photomicrography. B.

Fig. 6

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

Fig. 7

2) Opening the spring-loaded finger of the specimen holder with one hand, place a specimen slide inside the holder. (Fig. 8) When the slide comes in contact with the back of the specimen holder, slowly return the spring-loaded finger. WARNING: I f the spring-loaded finger is returned quickly, it may cause damage t o the specimen slide. Fig. 8

I Cover Glass I A n Olympus objective engraved "16010.17" requires a cover glass o f 0.17mm thickness. I f the numerical aperture o f the objective is 0.7 or higher (except immersion objectives) and no correction collar is provided, the resolving power deteriorates t o a great extent if cover glass thickness deviates from the above listed value. NOTE: I n some countries a 0.17mm cover glass corresponds t o a designation o f # I % . A cover glass (0.4mm thick) for blood counting, etc. can be used with Olympus objectives of the working distance longer than 0.4mm. (See VI. OPTICAL D A T A at page 16.)

1 Specimen Slide I Specimen slides 0.8 m m t o 1.2 m m thick are recommended for Olympus objectives.

3) Bring the portion o f the specimen for observation into the light path by means of the low drive control knobs. (Fig. 9)

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

The specimen holder can accommodate two standard specimen slides simultaneously. The specimen holder is removable t o obtain a large unobstructed stage surface to hold specimens up t o 5 5 mm x 8 5 mm.

Fig. 9

T o rotate the stage loosen the stage clamping screw @ and holding this screw, rotate the stage into the desired direciton. (Fig. 9 )

Q Stage clips@, for use with immersion objectives. (Fig. 10) A pair of stage clips are optionally available t o 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 o n the specimen holder. Fig. 10

C.

.-=,-.-.

Observation Tube

-

1. 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)

Fig. 11

- ----v ---z *

!-=F-

2. 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

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

-

. - r "

Fig. 13

Indicator Plate

I Amount of light

Application

I

Knob Position

I / I

Pushed in all the way (V)

I

Pulled out halfway (C. V)

Pulled out all the way

(C)

20% into binocular tube 100% into binocular tube 80% into photo tube 1100% into photo tube

@ Observation @ Dark specimens

@ Observation of exces-

Photomicrography of sively bright specimens dark specimens @ Photomicrography (focusing through the binocular tube)

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) C .V: C,amera & 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 by rotating in the direcknurled ring tion of the arrow. (Fig. 14) 2) Use the condenser height adjustment knob @ t o 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.

Fig. 15

Fig. 14

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. I f the polygonal image of the iris diaphragm becomes inscribed in the field i t means that the field diaphragm is centered. (Fig. 15)

* If

the specimen slide used is thicker than 1.2mm, you cannot obtain the image of the field iris diaphragm. Especially for photomicrography, use a specimen slide 0 . 9 m m to 1.2mm as much as possible.

1 Field lris Diaphragm ] The field iris diaphragm controls the diameter of the ray bundle impinging on the specimen surface and therefore, by stopping down the field diaphragm until it is slightly larger than the field o f view, it can reduce stray light, which in turn increases image definition and contrast. Aperture lris Diaphragm 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, t o 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. I 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 iris diaphragm can be seen in the objective pupil. Adjust the opening of the diaphragm to match the N.A. of the objective in use. If the specimen is low in contrast, it is recommended to stop down to 70% t o 80% of the objective N.A. (Fig. 16)

Fig. 16

E.

Focus Adjustment

1. Tension of Coarse Adjustment Knobs and Fine Adjustment.

Although the tension of the coarse adjustment knobs has been already adjusted for optimum performance by the manufacturer, it is possible to personally adjust the tension of the coarse adjustment for either heavy or light movement depending on the operator's preference b y 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. 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.

[ Use of Rubber Cap for Fine Adjustment Knob I t

Attaching this cap over the fine adjustment knob increases the sensitivity of the fine focusing motion. (The rubber cap i s 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. 18

3. Adjustment of Stage Block Height In addition t o 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 thicker than standard slides, e.g. chambers, flasks, etc. with much larger thickness. The stage block height can be adjusted by loosening the stage block locking screw @ with the Allen wrench provided and retightening it at the lower position. (Fig. 19)

Fig. 19

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 of the immersion objective.

3) Turn the revolving nosepiece to 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.

@ After use carefully wipe off the immersion oil deposited on the lens surfaces with gauze moistened with xylene. Never leave oil on the lens surfaces after 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 to be used with the BHS microscope for routine and advanced photomicrography. A separate, detailed instruction manual is available for the PM-IOAD camera system. For quick reference, however, you may want to refer to the follow~ngpointers when using the PM-IOAD. 1. Photographic Eyepiece Use NFK photo eyepieces for photomicrography. Insert the eyepiece @) into the eyepiece tube of 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 to the photo tube. (Fig. 21)

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

Fig. 21

4. Color and B & W Photomicrography a) Color photomicrography Voltage setting... Within a range as marked with the letters "PHOTO" on the voltmeter. Filter... Insert the LBD-2 filter into the filter mount at the light exit on the microscope base. (in case of daylight film) Light intensity... Should be adjusted by means of the ND filters rather than by means of voltage change. The range of voltage setting mentioned above indicates a guide line for color photomicrography. I f the photograph, taken this way, however, gives bluish tint, lower the voltage slightly, and i f i t gives reddish tint, raise the voltage slightly. b) Use of contrast filters for B & W photomicrography, @ Green filter... This filter is most recommended, since it aids i n obtaining photomicrographs of highest resolution and maximum contrast. @ Yellow filter... Recommended as contrast filter when i t is desirable t o emphasize the blue color in the stained portion of a specimen. @ Cobalt filter... Not generally recommended for photomicrographic purposes since it reduces both resolution and contrast. Use it for specific applications only when it is desirable to emphasize yellow in the stained portion of a specimen. @ No filter... A photograph is taken with less contrast than with thegreen filter.

5. Focusing Procedure Use the field of view eyepieces for focusing on the film 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 NF K photo eyepeice. (Fig. 22). The number at each frame indicates the magnification of the photo eyepiece. The c,s71 image in the field of view eyepiece and the image on the film plane are in focus at the L & , J J ~ ~ same time. Several type field of view eyepieces are available, according to the, film size employed. Fig. 22

<_:

Field of view eyepiece 35WH K10X

PWH K1OX

4X5WHKlOX

MHWH K1OX

35mm Back

3%" x 4%" Polaroid Back

4" x 5" Sheet Film or Polaroid Film Holder

16 mm Bolex camera 120 Roll Film Holder

Attachment camera

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 t o be photographed within the frame corresponding t o 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) It is recommended to tighten the tension adjustment ring considerably t o prevent the stage from dropping during long exposures.

VI. OPTICAL DATA

WHK1OX (Field number 20'

Focal length 34.23

17.69

8.99

4.61

1.75

36.54

18.98

8.03

4.13

1.69

3.36

1.34

0.84

0.52

0.27

2.58

1.12

0.73

0.48

0.27

40X

lOOX

200X

400X

lOOOX

40X

lOOX

200X

400X

lOOOX

173

27.6

9.15

3.0

0.68

125.4

22.1

7.6

2.72

0.68

5

2

1

0.5

0.2

5

2

1

0.5

0.2

Totalmag. depth (PI Field of view (mm)

Immersion objectives The resolving power and focal depth are obtained with fully opened aperture diaphragm. Technical terms: Working distance:

The distance from the cover glass to the nearest point of the objective.

Numerical aperture:

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

Resolving power:

The ability of a lens to register small details. The resolving power of a lens is measured by its ability to separate two points.

Focal depth:

The distance between the upper and lower limits of 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. of an objective the shallower the focal depth.

Field number:

A number that represents the diameter in mm of the image of the field diaphragm that is formed by the lens in front of it.

Field of view diameter: The actual size of the field of view in mm on the object surface.

VII. TROUBLESHOOTING GUIDE I f you are unable to obtain full performance from your microscope, please consult with the table below as pointers for troubleshooting.

Phenomenon

Cause

Remedy

Field iris diaphragm is not opened sufficiently.

Open diaphragm to proper diameter.

Condenser much.

Adjust condenser height.

1. Optical System a) With illuminator switched on, the field of view is dark.

b) Field of view iscut off or illuminated irregularly.

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

is lowered too

Light path selector lever is pulled out to C position.

Push in lever up t o CV or V position.

Light path selector lever is stopped midway.

Click it into proper position according to your purpose.

Nosepiece is not clicked into place.

Slightly rotate nosepiece until it clicks into place.

Nosepiece is not correctly mounted.

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

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

Choose a condenser t o meet your purpose.

Condenser is not centered.

Center condenser.

Field iris diaphragm is stopped down excessively.

Open diaphragm to proper diameter.

Dust, etc. on light exit lens.

Remove dust, etc. 'lean front lenses-

Dust on condenser top lens. Dirty specimen. Dust on eyepiece.

d l Excessive image contrast.

Condenser much.

is lowered too

Aperture iris diaphragm is stopped down excessively.

Adjust condenser height. Open diaphragm to proper diameter.

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

Cause

Remedy

Non Olympus objectives are used.

Use Olympus LB seriesobjectives.

Nosepiece is not correctly mounted.

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

Objective is not correctly positioned in the light path.

Click nosepiece into place.

Objective correction collar is not adjusted.

Rotatecorrectioncollar, keeping specimen in fine focus until optimum resolution is obtained.

Dust on objective front lens.

Clean front lens.

lmmersion objective is not used with immersion oil.

Use immersion oil.

Bubbles in immersion oil.

Remove bubbles (and reapply oil).

lmmersion oil designated by Olympus is not used.

Use Olympus immersion oil.

Dirty specimens. 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.

Clean.

Nosepiece is not correctly mounted.

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

Objective is not correctly positioned in the light path.

Slightly rotate nosepiece until it clicks in place.

Specimen is not correctly positioned on stage.

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

Nosepiece is not correctly mounted.

Insert 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 much.

Adjust condenser height.

is lowered too