BHTU BIOLOGICAL MICROSCOPE Instructions March 1993

CONTENTS I.

STANDARD CONFIGURATIONS

2

II.

NOMENCLATURE

3

III.

ASSEMBLy... ...

4

IV.

IDENTIFICATION AND FUNCTION OF VARIOUS COMPONENTS

6

V.

OPERATION

9

A.

9

Switching ON the Light Source

I Voltage Adjustment and Light Intensity I I LP (Light Intensity Presetting) Switch I B.

10

Placement of a Specimen Slide .

I White Cover for Condenser

I Cover Glass I I Specimen Slide I I Stage I C.

Observation Tube 1. Interpupillary Distance Adjustment 2. Diopter Adjustment 3. Light Path Selector

D.

13

Condenser Adjustment 1. Condenser Centration

I Filed Iris Diaphragm I I Aperture Iris Diaphragm E.

Focusing Adjustment

.

14

1. Tension of Coarse Adjustment Knobs and Fine Adjustment

I Use of Rubber Cap for Fine Adjustment Knob 2. Pre-Focusing Lever F.

Use of Immersion Objectives

G.

Photomicrography

15

.

15

VI.

OPTICAL DATA

17

VII.

TROUBLESHOOTING GUIDE

18

"",

I.

STANDARD CONFIGURATIONS Model Component

BHTU·ll1 BHTU-112 BHTU·312 Microscope stand

BHTU·F

1

1

1

8 inocular tube

BH2·B130

1

1

0

Trinocular tube

BH2·TR30

0

0

1

1

1

1

with quintuple revolving nosepiece Observation tubes

Square mechanical stage with right-hand low BH2.SVR2 drive coaxial controls and specimen holder Swing-out condenser

BH2·SC

1

1

1

Halogen lamp holder

LS·20H

1

1

1

Halogen bulbs

6V20WHAL

2

2

2

D Ach. 4X, D Ach. lOX, D Ach, 40X, D Ach. lOOX (oil)

each

1

0

0

Objectives

D Plan 4X, 0 Plan lOX, D Plan 40X D Plan lOOX (oil)

0

1

1

each

each

Eyepieces, paired

WKlOX

1

1

1

Photo eyepiece

NFK3.3X

0

0

1

Filter

KB·4

1

1

1

Immersion oil, bottled

1

1

1

Vinyl dust cover

1

1

1

2

II.

NOMENCLATURE

The Model BHTU 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.

Eyepiece

-

Observation tube

Micoscope stand For biological use

Objective

Stage

Halogen lamp holder

-- --

Condenser

LP (Light Intensity Presetting) switch

3

III.

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 :;cratching the glass surface. For numbers ~ and f) please refer to explanations in detail on the next page.

NOTE:

Eyepjece

0)

.~

®

.g Objective

Observation tube

Circular dovetail mount

Specimen

holder

Observation tube clamping screw

Clamping

screws

Condenser clamping screw

~tage L'_I...;~;. -

Halogen bulb

Stage mounting dovetail

e

CD,Q

~ Halogen lamp

®

9J

holder

Condenser (The N.A. scale engraved on the con-

denser shou ld face the

-.. To AC ® outlet

microscope front.)

LP (Light Intensity Presetting) switch

4

• Explanations in detai I

9

Mounti ng the stage 11 Loosen the stage clamping screw CD by rotating counterclockwise. (Fig. 1) 2) Insert the stage into the mounting dovetail of the microscope stand slowly and lock with clamping screw.

Fig.

fJ Mounting the observation tube 1) Loosen the clamping knob CD fully. (Fig.21 Pull spring-loaded clamping knob CD. This will cause the locating pin <V to· withdraw. If the pin does not, loosen the screw further until the pin withdraws. 2) With clamping knob CD pulled out. insert the circular dovetail of the observation tube into the ring dovetai I. 3) Tighten the clamping knob.

5

Fig. 2

Diopter adjustment ring

Photo tube

Condenser height adjustment knob

Pre- focusi ng lever

X-axis low drive control knob X excursion range: 76 mm.

Y-axis low drive control knob Y excursion range: 50mm.

Field iris diaphragm ring Arrow mark (0 ... 0 indicates increase in diaphragm diameter.

Line cord

7

I

Summary of Putting the Microscope into Operation

II

MODEL BHTU A.

Match the voltage selector switch to local mains voltage (page 9).

B.

Switch on the light source (page 9).

C.

Place a specimen slide on the mechanical stage.

O.

Coarse focus with the lOX objective.

E.

Make interpupillary distance and diopter adjustments (page 12).

F.

Adjust the condenser position (page 13).

G.

Swing in the desired objective.

H.

Adjust light intensity.

I.

Fine focus.

J.

Adjust aperture iris diaphragm and field j'ris diaphragm ~page 13).

I Adjustment of Illumination System for Various Objective Powers II Condenser Objective magnification

Achromaticaplanatic

condenser BH2-AAC

Abbe condenser

Swing·out condenser

Low power condenser

BH2-CD

BH2-SC

BH2-UL-C

Swi ng out top lens

Compatible

lX 2X 4X lOX 20X 40X

Compatible Compatible

BOX

Swing in top lens

----------.-

•

lOOX

*N.A. is somewhat low, but still compatible with a 100X objective.

(Take a copy of this page and put it on the wall near the microscope for use as a reminder of microscopic procedure.)

OLYMP'US® 8

v.

OPERATION

A.

Switching on the Light Source

1) Ascertain that the voltage selector SWitch CD

is set to conform WIth the local mains voltage. (Fig. 3) If the switch is not correctly set. adjust it by

means of the Allen wrench provided, or a screw-driver.

2) Place the sliding voltage control lever on the nght side of the microscope base to a lXlSition dosest to you (low voltage position). Switch on the light source (ma;n SWitch (2)1. (FIQ. 3)

,. Fig. 3

IVoltage Adjustment and Ugh! Intensity I As you push the control lever (j) in the direction of the ARROW. the voltage increases. and the LED readout (2) ¥ill display the lamp voltage. (Fig. 4)

IlP (light Intensity Presening) Switch Apart from the light intensity set by voltage adjustment control lever, the LP switch can set the light intensity as required.

Fig. 4

Presetting the light intensity (Fig. 5) 1) Switch ON the LP switch CD.

2) Insert a slotted screwdriver into me LP adjust~ ment screw (1). Turning it clockwise will increase the light intensity. Turn it until required intensity is obtained. The light intensity will not change even if the voltage adjustment lever @ with the LP switch ON. To change the light intensity by using the voltage adjustment lever, switch the LP switch OFF. To recall the light intensity preset, push the LP switch to ON.

*

A Hint on Convenient Application of the LP Switch For instance, when you want to use two kinds of objective lens ahernatley. set one objective by using the vohage adjustment [ever. then set another objective lens by using the LP switch. This allows the ffght intensity 10 recallhe intensity initially preset by switching the LP swilch ON and OFF. This also allows the light intensity setting for photomicrography.

9

Fig. 5

B.

Placement of a Specimen Slide

11 Rotate the coarse adjustment knobs CD in the direction of the ARROW to rack down the stage so that a specimen slide can be placed on the stage. (Fig. 6) 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 with the other hand. When the slide comes in contact with the back of the specimen holder, slowly return the springloaded finger.

.

..

.~~

~~ ... -

~

/~-"'"'"

Fig. 6

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

3) For use of the specimen holder B2-HL2 A specimen slide can be easily slipped into the specimen holder by single hand operation. (Fig.,

71 Fig. 7 White Cover for Condenser The white cover for condenser CD should be covered over the condenser. (Fig. 8) • When setting stain specimens under the specimen holder, the white cover for condenser comes into field of view as a background. This allows easy distinction what you are observing between its background.

10

ICover Glass I •

An Olympus objective engraved "16010.17" 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 greatly, if cover glass thickness deviates from the above listed value. NOTE:

•

In some countries a 0.17 mm thick cover glass corresponds to a #1 Y, cover glass.

A cover glass (0.4 mm thick) for blood counting, etc. can be used with Olympus objectives except 0 Plan 40X. S Plan Apo 40X and S Plan lOOX.

ISpecimen Slide I •

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 dark field condenser and the dj fferential interference contrast con~ denser.

3) Bring the portion of the specimen to be

observed into the light path by means of the low drive stage control knobs. (Fig. 9)

* Tighten the stage clamping screw <D .

IStage I •

The specimen holder can accommodate two standard specimen slides simultaneously.

•

The specimen holder is removalbe to obtain a large unobstructed stage surface to hold specimens up to 55 mm x 85 mm.

•

To rotate the stage loosen the $tage clamping screw CD and holding this screw, rotate the stage into the desired direciton. (Fig. 10)

Fig.10

@ Stage clips for use with immersion objectives.

(Fig. 111 A pair of stage clips is optionally available to hold the specimen on the stage, eliminaing specimen drag or buckling due to immersion oil between slide and stage surface. The cI ips can be inserted into the holes CD provided on the specimen holder.

Fig. 11

11

c.

-

Observation Tube

1. Interpupillary Distance Adjustment 1} Click the lOX 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 CD of the right and left eyepiece tubes with b9th hands until perfect binocular vision is obtained. {Fig. 121

Fig. 12

2. Diopter Adjustment 1) Looking at the image through the right

eyepiece with your right eye, focus on the specimen with the fine adjustment knobs. 21 Next, looking at the image through the left eyepiece with your left eye, rotate the diopter adjustment ring CD to focus on the specimen without using the coarSE and fine adjustment knobs .. (Fig. 131

~J

tii

&,~ :t

3. Light Path Selection

./11

- ,,~ 'i~

CD

1) The trinocular tube is provided with a light path selector lever CD to direct the light to the observation tube and/or to the photo tube in 3 positions. (Fig. 141

,1lIII,' ~

~

[1

.~

_f'~"

r

-.:.$

'

Fig. 14 Indicator Plate Lever Position

Amount of light

Applica· tion

Pushed in all the way

Pulled out: halfway

Pulled out all the way

(VI

(C. 11.1

(C)

100% into binocular tube

20% into binocular tube 80% into photo tube

100% into photo tube

CD Observation

CD Observation of exces-

CV Dark specimens

Photom icrography of sively bri~lht specimens dark specimens ~ Photomicrography (fo· cusing through the binocular tube)

An indicator plate is provided at the knob port to summarize the usage of the above table; it can be consulted before operating the knob V: Viewer (white letters) CV: Camera & viewer (yellow-green letters) C: Camera (red letters) The colors of the letters correspond wi th the color bands on the knob shaft. 12

D.

Condenser Adjustment

1. Condenser Centration 1) Stop down the field iris diaphragm with knurled ring CD 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

®

Fig. 14

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 it means that the field diaphragm is centered. (Fig. 15) Field Iris Diaphragm •

I

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 it is slightly larger than the field of view, it can reduce stray light, which in turn increases image definition and contrast.

Aperture Iris Diaphragm

I

•

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. It 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 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% - 80% of the objective N.A. (Fig. 161

13

70-80% 20 - 30"," Opening of the aperture diaphragm Objective exit pupil

Fig. 16

E

Focus Adjustment

CD

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 by rotating the tension adjust-

f

Fig. 18 ment ring <D. (Fig. 181 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, 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 ill is provided to prevent possible contact between specimen and objective as well as to simplify coarse focusing. (Fig. 191 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 the stage is lowered and then raised again. The pre-focusing lever does not restrict fine focusing.

14

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) Rotate the revolving nosepiece to bring the immersion objective into the light path, and focus with the fine adjustment knobs. NOTE: ill 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 to prevent oil bubbles from forming in the oil film between condenser and specimen slide. If any bubbles appear, 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 perforrrance of the lens system.

,

G.

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

1. Photographic Eyepiece Use NFK photo eyepieces for photomicrography. Insert the eyepiece CD 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- lOAD body and clamp the camera unit to the photo tube. (Fig. 211

3. Setting the Light Path Selector Refer to section C.3. on page 12.

15

4. 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 relide with 4 frames, each frame indicating the area covered by a specific power N FK photo eyepeice. (Fig. 22) The number at each frame indicates the magnification of the photo eyepiece. The image in the field of view eyepiece and the image on the film plane are in focus at the same time. Several type field of view eyepieces are available, according to the film size employed. Fig. 22 Field of view eyepiece

Attachment camera

35WHK10X

PWHK10X

4X5WHK10X

MHWHK10X

35mm Back

3%" x 4%" Polaroid Back

4" x 5" Sheet Film or Polaroid Film Holder

16mm Bolex camera 120 Roll Film Holder

1) Select the field of view eyepiece matching the camera bSlck in use and insert it 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 on account of their considerable depth of focus. 3) Bring the specimen detail to be photographed within the frame corresponding to the power of the NF K 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 yellowijreen (eVI band. 4) It is recommended to tighten the tension adjustment ring considerably to prevent the stage from dropping during long exposures.

16

VI.

OPTICAL DATA Objective

Magnifi-

4X

lOX

40X

l00X'

4X

lOX

40X

100X'

N.A.

0.10

0.25

0.65

1.30

0.10

0.25

0.65

1.25

W.O. (mm)

18.2

7.2

0.6

0.20

7.03

7.4

0.27

0.17

30.03

16.9

4.58

1.91

34.23

17.5

4.67

1.75

Resolving power (jJ)

3.36

1.34

0.52

0.26

3.36

1.34

0.52

0.27

Total mag.

40X

100X

400X

1000X

40X

100X

400X

lQOOX

Focat depth {jJJ

171.6

27.45

3.0

0.7

171.6

27.45

3.0

0.7

Field of view (mm)

5

2

0.5

0.2

5

2

0.5

0.2

cation

Focal length

lmml Eyepiece

WK10X (Field number

201

o Plan Ach.

o Achromat

Type

ll-Immersion objectives The resolving power and focal depth are obtained witt' 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. Th~ N.A. values can be used for directly comparing the resolving

powers of all types of objectives. The larger the NA. the higher 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 yo u 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.

17

VII. TROUBLESHOOTING GUIDE If you are unable to obtain full performance from your microscope, please consult with the

table below as pointers for troubleshooting.

Phenomenon

Remedy

Cause

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

b) Field of view is cut off or

illuminated irregularly.

Field iris diaphragm opened sufficiently. Condenser much.

is

IS

lowered

not

too

Adjust condenser height.

Light path selector lever is pulled out to C position.

Push in lever up to CV or V position.

Light path selector lever is

Click it into proper position according to your purpose.

stopped midway. Nosepiece is not clicked into

In

diaphragm to proper

diameter.

place.

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

Open

Slightly rotate nosepiece until it cl icks into place. Choose a condenser to meet

The power of objective used exceeds the illu'mination capacity of condenser.

your purpose.

Condenser

is not centered.

Center condenser.

Field irisdjaphragm is stopped down excessively.

Open diaphragm diameter.

Dust, etc. on light exit lens

Remove dust, etc. Clean front tenses.

Dust on condenser top lens

to proper

Dirty specimen Dust on eyepiece d) Excessive image contrast

Condenser much.

is

lowered

Aperture iris diaphragm stopped down excessively.

18

too is

Adjust condenser height. Open diaphragm to diameter.

proper

Phenomenon el Resolution problems:

Remedy

Cause

LB objectives series are used.

other

than

• Image is not sharp.

o bjectiv8 IS not correctly • Insufficient contrast. I positioned in the light path. • Image details lack defi- Objective correction collar is nition.

not adjusted correctly.

1

Use Olympus LB series objectives. Click nosepiece into place. Rotate correction collar, keeping specimen in fine focus until optimum resolution is obtained.

Dust on objective front lens

Clean front lens.

Immersion objective is not used with immersion oil.

Use immersion oil.

Bubbles in immersion oil

Remove bubbles (and reapply oil).

Immersion oil designated by Olympus is not w~ed.

Use Olympus immersion oil.

Dirty specimens Clean. Dust on condenser lens fl Field of view is partially out of focus, or image is partly out of focus.

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.

gl Specimen image is partialIy out of focus.

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

Objective is not correctly positioned in the light path.

Slightly rotate nosepiece until it clicks into place.

Condenser is not centered.

Center condenser.

Condenser centered.

is

not correctly

Center condenser.

Condenser much.

is

lowered

Adjust condenser height.

too

2. Electric System al Illuminator is too bright (or too dark) even when adjusting control lever.

Line voltage selector switch is not matched with local mains voltage.

Match selector mains voltage.

switch

to

b) Voltage for illuminator cannot be raised. cl Lamp goes off and on.

Bulb filament burn out. Loose

d) Bulb burns out frequently.

is likely

to

electric connections.

Replace bulb. Check all connections.

Line voltage selector switch is not matched with local mains voltage.

Match selector mains voltage.

Bulb is not standard one.

Use standard bu lb.

19

switch

to

Phenomenon

Remedy

Cause

3, Coarse and Fine Adjustments adjustment knob is Tension adjustment ring is Loosen ring properly. I aj Coarse too tight. tightened too much. Users is trying to raise stage above the focusing limit with pre-focusing lever engaged.

bl Stage drops or specimen goes out of focus during observation due to slipping fine adjustment knobs. c) Stage cannot be raised to the upper limit.

Tension adjustment too loose.

ring

is

Pre-focusing lever is engaged

Unlock pre-focusing lever.

Tighten ring properly.

Unlock lever.

in lower than focusing position.

dl Stage cannot be lowered to the lower limit.

e) Objective front lens hits specimen before into focus.

coming

Condenser mount is lowered too much.

Raise condenser.

Specimen is placed on stage upside down ..

Reverse specimen.

Interpupillary distance is not correctly adjusted.

Correct the interpupillary distance.

4, Observation Tubes

al Incomplete binocular visian.

Diopter adjustment is incom-

Complete the diopter adjust-

plete,

ment.

Right and left eyepieces are

Use a pair of matched eyepieces.

not matched. User is unaccustomed to binocular vision.

Prior to looking into the binocular observation tube, look at a far away object.

Stage is not correctly locked.

Clamp stage securely.

Specimen is not correctly positioned.

Adjust specimen pOSitioj

5, Stage

a) Image easily goes out of focus when you touch the stage.

bl Specimen

stops midway on the east-west traverse.

20