Optika Italy B-1000 Series User manual

Ver. 2.1 2020
Model
B-1000FL-LED
B-1000 Series
INSTRUCTION MANUAL

Page 2
Table of Contents
1. Warning 3
2. Symbols and conventions 3
3. Safety Information 3
4. Intended use 3
5. Instrument description 4
6. Unpacking 6
7. Assembling 6
7.1 Assembling the microscope 7
7.1.1 Manual version 7
7.1.2 Motorised version 9
8. Brighteld (transmitted light) observation procedures 10
9. Use of the microscope in Brighteld (transmitted light) 11
9.1 General switch on 11
9.2 Control keyboard 11
9.3 Brightness adjustment 11
9.4 Adjust the observation head 12
9.5 Adjust the interpupillary distance 12
9.6 Diopter adjustment 12
9.7 Use of eyeshields 12
9.8 Light path selection 13
9.9 Coarse focus tension adjustment 13
9.10 Focus stop lever 14
9.11 Stage 14
9.12 Centering the condenser 15
9.13 Effect of eld diaphragm 15
9.14 Aperture diaphragm 15
9.15 Use of oil immersion objective 16
9.16 Only for motorised version 17
9.16.1 Nosepiece rotation 17
9.16.2 Focusing 17
9.16.3 Stage 17
10. Fluorescence (reected light) observation procedures 18
11. Use of the microscope in Fluorescence (reected light) 19
11.1 LED switch on 19
11.2 Use of uorescence 19
11.3 Use of light excluding plate 20
12. Microphotography 21
12.1 Use of “C”mount camera 21
12.2 Use of Reex camera 21
13. Maintenance 22
14. Troubleshooting 23
Equipment disposal 25

Page 3
1. Warning
This microscope is a scientic precision instrument designed to last for many years with a minimum of
maintenance. It is built to high optical and mechanical standards and to withstand daily use. We remind you
that this manual contains important information on safety and maintenance, and that it must therefore be made
accessible to the instrument users. We decline any responsibility deriving from incorrect instrument use uses
that does not comply with this manual.
2. Symbols and conventions
The following chart is an illustrated glossary of the symbols used in this manual.
CAUTION
This symbol indicates a potential risk and alerts you to proceed carefully.
ELECTRICAL SHOCK
This symbol indicates a risk of electrical shock.
3. Safety Information
Avoiding Electrical Shock
Before plugging in the power supply, make sure that the supplying voltage of your region matches with the
operation voltage of the equipment and that the lamp switch is in off position. Users should observe all safety
regulations of the region. The equipment has acquired the CE safety label. However, users have full responsibility
to use this equipment safely. Please follow the guidelines below, and read this manual in its entirety to ensure
safe operation of the unit.
4. Intended use
Standard models
For research and teaching use only. Not intended for any animal or human therapeutic or diagnostic use.
IVD Models
Also for diagnostic use, aimed at obtaining information on the physiological or pathological situation of the
subject.

Page 4
COARSE
FOCUSING
KNOB
NOSEPIECE
DIOPTRIC
ADJUSTMENT
RING
PHOTO/TV
PORT
MAIN SWITCH
EYEPIECE
FILTER
SELECTOR
LEVER
CONDENSER
CENTERING
KNOBS
TRANSMITTED
LIGHT BRIGHTNESS
ADJUSTMENT KNOB
FOCUS LOCK
LEVER
FINE FOCUSING
KNOB
OBJECTIVE
STAGE
FLUORESCENCE LED
5. Instrument description
CONTROL
KEYBOARD

Page 5
FIELD DIAPHRAGM
APERTURE
DIAPHRAGM
Opposite side
FLUORESCENCE
MAIN SWITCH /
BRIGHTNESS
ADJUSTMENT
KNOB
LIGHT PATH
SELECTOR
LEVER
TENSION
ADJUSTMENT
RING
CONDENSER
HEIGHT
ADJUSTMENT
KNOB
X-Y STAGE
MOVEMENT
KNOBS

Page 6
①
②
③
④
⑤
⑥
⑦
⑧
6. Unpacking
The microscope is housed in a moulded Styrofoam container. Remove the tape from the edge of the container
and lift the top half of the container. Take some care to avoid that the optical items (objectives and eyepieces)
fall out and get damaged. Using both hands (one around the arm and one around the base), lift the microscope
from the container and put it on a stable desk.
Do not touch with bare hands optical surfaces such as lenses, lters or glasses. Traces of grease or
other residuals may deteriorate the nal image quality and corrode the optics surface in a short time.
7. Assembling
Once opened the box, the microscope parts are the following:
① Frame
② Objectives
③ Stage
④ Condenser
⑤ Observationhead
⑥ Eyepieces
⑦ LEDuorescenceattachment
⑧ Lightexcludingplate
⑨ Dustcover
⑩ Allenwrench
⑪ Immersionoil(if100xobj.isincludedinthe
 conguration)
⑫Powersupply(2units)
⑪
⑩
⑨
⑫

Page 7
7.1 Assembling the microscope
7.1.1 Manual version
1. Put the microscope stand on a solid table. Insert
uorescence attachment above the stand, using
the Allen wrench to tighten the screw. (Fig. 1)
2. Insert the optical head above the attachment, us-
ing the Allen wrench to tighten the screw. (Fig. 2)
3. Insert eyepieces into the empty eyepiece sleeves.
(Fig. 3)
4. Insert the condenser under the stage: position
until it is well inserted into its holder (under the
condenser there is a pin that must fully enter the
holder guide). (Fig. 4)
5. Lock the condenser xing knob ①.
FFig. 1ig. 1
FFig. 2ig. 2
FFig. 4ig. 4
FFig. 3ig. 3
①

Page 8
FFig. 5ig. 5
FFig. 6ig. 6
FFig. 8ig. 8
FFig. 7ig. 7
6. Mount the stage: lower the support using the
coarse focus knob, then place the stage and
tighten the lock screw ①. (Fig. 5)
7. Screw each objective into the thread of the nose-
piece, clockwise with increasing magnication.
(Fig. 6)
8. Insert the power supply jack in the socket placed
in the rear side of the of the microscope: one for
transmitted light and one for uorescence. (Fig.
7-8)
①

Page 9
FFig. 9ig. 9
FFig. 10ig. 10
FFig. 11ig. 11
②
①
③ ④ ⑤⑥
7.1.2 Motorised version
1. Assemble the stage in the same way as the
manual version. Check the perfect alignment of
the rear part of the stage with the rear arm of the
frame. An imperfect alignment could lead to an
incorrect functioning of the system. (Fig. 9)
2. Connect the cable ① from the stage to the frame
and tight the locking screws of the connectors ②.
(Fig. 10)
3. Connect the provided cables: ③ 12V power sup-
ply for the motorised parts; ④6V microscope
power supply; ⑤ serial cable; ⑥ PS/2 mouse.
(Fig. 11)
• Connect power cables as the last step.

Page 10
8. Brighteld (transmitted light) observation procedures
Main switch
(Used commands) (Chapter)
Slide holder
Revolving nosepiece
Observation head
Diopter adjustment ring
Condenser centering
Aperture diaphragm
Field diaphragm
Revolving nosepiece
Light adjustment dial
Coarse and fine focusing knobs
Coarse and fine focusing knobs
Bring on “1” the main switch
Put a specimen on the stage
Insert 10X objective into the light path
Focus the specimen
Begin observation
Adjust field and aperture iris diaphragms
Insert desired objective for the observation and focus the
specimen
Adjust interpupillary distance
Adjust diopters
Center optical axis
Adjust light intensity
9
9
5
5
5
5
9
9
9
9
9
9
ON-OFF key on the keyboard
Light adjustment dial
Turn on the microscope and adjust light intensity 9
9
Fluorescence filter selector
Remove all fluorescence filter from the light path 11

Page 11
9. Use of the microscope in Brighteld (transmitted light)
9.1 General switch on
1. To activate the transmitted light illuminator, put
the main switch ①, located on the left side of the
stand, to the position “1”. (Fig. 12)
9.2 Control keyboard
B-1000 illumination can be managed through the
keyboard placed on the left of the stand. (Fig. 13)
• ON-OFF (②): press this key (after switching the
main switch on “1”) to turn ON or OFF the micro-
scope LED.
• BOOST (③): press this button in order to in-
crease the brightness (useful for high-magni-
cation objectives or very opaque specimens).
Do not enable boost mode while observing
with low magnication objectives (4x, 10x)
with fully open diaphragm: the high bright-
ness may hurt user’s eyes.
• AUTO OFF (④): if you want the illuminator to
switch off automatically, press this button until 15,
30 or 60 minutes delay is set. After this period of
time, the light will turn off. You have to press the
ON-OFF button to turn it on again.
9.3 Brightness adjustment
1. Use the brightness adjustment dial ⑤ on the left
side of the microscope to increase or decrease
the light intensity on the specimen. (Fig. 14)
③
④
②
FFig. 13ig. 13
FFig. 14ig. 14
⑤
①
FFig. 12ig. 12
①

Page 12
FFig. 15ig. 15
FFig. 16ig. 16
FFig. 18ig. 18
FFig. 17ig. 17
①
③
②
9.4 Adjust the observation head
1. Loosen the locking screw ①, turn the observation
head to a comfortable position for observation,
and then lock the locking screw again. (Fig. 15)
9.5 Adjust the interpupillary distance
1. Observing with both eyes, hold the two eyepiece
prism assemblies. Rotate them around their com-
mon axis until the elds of view coincide.
• The graduation on the interpupillary distance
indicator ②, pointed by the spot “.” on the
eyepiece holder, shows the distance between
the operator’s eyes. (Fig. 16)
The range of the interpupillary distance is 48-75 mm.
9.6 Diopter adjustment
1. Look into the right eyepiece with your right eye
only, and focus on the specimen.
2. Look into the left eyepiece with your left eye
only. If the image is not sharp, use the dioptric
adjustment ring ③ to compensate. (Fig. 17)
• The adjustment range is ±5 diopter. The
number indicated on the adjustment ring
graduation should correspond to the
operator’s dioptric correction.
9.7 Use of eyeshields
• Use with eyeglasses
1. Fold rubber eyeshields with both hands. Folded
eyeshields avoid scratching the lenses of eye-
glasses. (Fig. 18)

Page 13
FFig. 19ig. 19
FFig. 21ig. 21
FFig. 20ig. 20
②
①
• Use without eyeglasses
1. Raise eyeshields and observe at the microscope
placing eyes to the shields, avoiding external light
to disturb the observation. (Fig. 19)
9.8 Light path selection
• The observation head is equipped with an optical
path selector that allows the light to be splitted to
the eyepieces and to the photo / TV port.
1. Move the selector ① to one of the three possible
positions to split the light. (Fig. 20)
POSITION LIGHT
IN 100% EYEPIECES
MIDDLE 50% EYEPIECES / 50% TV
OUT 100% TV
9.9 Coarse focus tension adjustment
The tension of the coarse focusing knob is factory
preset.
1. To modify the tension according to personal’s
needs, rotate the ring ② . (Fig. 21)
• Clockwise rotation increases the tension.
• If the tension is too loose, the stage could go
lower by itself or the focus easily lost after ne
adjustment. In this case, rotate the knob in order
to increase the tension .

Page 14
9.10 Focus stop lever
The upper limit knob has two functions: prevent the
contact between slide and objective and acts as
“focus memory”.
1. After focussing the specimen, pull the lever ①
toward the front of the microscope and lock it.
(Fig. 22).
• In this way the focus upper limit is set.
2. Now one can lower the stage with coarse focus
knob, replace the specimen and raise again the
stage up to the upper limit: specimen will be in
approximate focus and will need a ne adjustment
to get the proper focus..
• Fine focus movement is not affected by the
coarse focus lock.
• To unlock, move the lever in the opposite
direction to the one used for the locking.
• Two stoppers ②are inserted on the frame. DO
NOT REMOVE THE TWO STOPPERS.
9.11 Stage
Stage accepts standard slides 26 x 76 mm, thickness
1,2 mm with coverslide 0,17 mm. (Fig. 23)
It is possible to place two slides side by side on the
stage.
1. Open the spring arm of the slide holder ① and
place from the front the slide on the stage.
2. Gently release the spring arm of the slide holder.
• A sudden release of the spring arm could
cause the falling of the slide.
FFig. 23ig. 23
①
FFig. 22ig. 22
①
LOCK
UNLOCK
②

Page 15
9.12 Centering the condenser
1. Place the specimen on the stage, insert 10x ob-
jective into the light path and focus.
2. Insert the front lens of the swing-out condenser
①. (Fig. 24)
3. Rotate the eld diaphragm ring ② clockwise, to
fully close the diaphragm.
4. Rotate the condenser height adjustment knob ③
to focus the edges of the diaphragm.
5. Rotate the two centering screws ④to bring the
bright spot in the centre of the eld of view.
6. Gradually open the diaphragm. The condenser is
centred when the diaphragm image is symmetri-
cal to the eld of view.
7. In normal use, open the diaphragm until it circum-
scribes the eld of view.
9.13 Effect of eld diaphragm
Field diaphragm adjusts the illuminated area to ob-
tain a high contrast image.
Set the diaphragm according to the objective in use
until it circumscribes the eld of view, in order to elim-
inate unnecessary light to eyepieces. (Fig. 25)
9.14 Aperture diaphragm
• The Numerical Aperture (N.A.) value of the ap-
erture diaphragm affects the image contrast. In-
creasing or reducing this value one can vary res-
olution, contrast and depth of focus of the image
• With low contrast specimens set the numeri-
cal aperture value ⑤ (printed on the condenser
ring) to about 70%-80% of the objective’s N.A.
(Fig. 26). If necessary, remove one eyepiece and,
looking into empty eyepiece sleeve, adjust the
condenser’s ring in order to obtain an image like
the one in Fig. 27.
Example: with objective PLAN 40x / 0.65 set the
scale to 0.65 x 0.8 = 0,52
FFig. 40ig. 40
FFig. 27ig. 27
APERTURE
DIAPHRAGM
FIELD OF VIEW
30-20%
70-80%
FFig. 25ig. 25
FFig. 26ig. 26
FFig. 24ig. 24
②
③
①④
⑤

Page 16
9.15 Use of oil immersion objective
1. Focus the specimen with a low power objective.
2. Lower the stage (remember to lock the coarse
upper limit knob).
3. Put a drop of oil (provided) on the area of the
specimen to be observed. (Fig. 28)
• Make sure that there are no oil bubbles. Air
bubbles in the oil damage the image quality.
• To check for bubbles: remove an eyepiece, fully
open the aperture diaphragm and observe the
objective exit pupil. (The pupil must be round and
bright).
• To remove the bubbles, gently move the nose-
piece to the right and to the left to move the im-
mersion objective a few times and allow the air
bubbles to move away.
4. Insert immersion objective in the light path.
5. Return the stage to the upper focusing point and
obtain an optimal focus using the ne focus knob.
6. After use, gently remove the oil with a soft pa-
per towel or a lightly moistened optic paper with a
mixture of ethyl ether (70%) and absolute alcohol
(30%).
• Immersion oil, if not immediately cleaned,
could crystallize creating a glass-like layer.
In this situation the observation of the speci-
men would be difcult if not impossible due to
the presence of an additional thickness on the
objective.
FFig. 28ig. 28

Page 17
FFig. 30ig. 30
FFig. 29ig. 29
②
①
9.16 Only for motorised version
9.16.1 Nosepiece rotation
1. To change magnication it is possible operate
on the nosepiece movement buttons located on
the right side of the frame. (Fig. 29). Orange but-
ton ① rotates the nosepiece clockwise, while the
blue button ② rotates the nosepiece counter-
clockwise.
2. As an alternative it is possible operate on the right
and left mouse buttons.
9.16.2 Focusing
• Focus motor is activated through the mouse
wheel.
1. Front or rear rotation of the mouse wheel raises
or lowers the stage. (Fig. 30)
9.16.3 Stage
1. Stage is moved through the mouse movement.
A mouse movement to the front or to the back ③
causes a stage movement of the stage along the
Y axis, while a left or right movement ④ causes
a stage movement of the stage along the X axis.
(Fig. 31)
• It is always possible, however, operate on the
translation knobs of the stage for a manual move-
ment.
FFig. 31ig. 31
③
④
③
④

Page 18
10. Fluorescence (reected light) observation procedures
Fluorescence main switch
(Used commands) (Chapter)
Slide holder
Fluorescence lter turret
Turn the fluorescence main switch to turn on the LED
Place a specimen on the stage.
Insert in the light path the filtercube
suitable for the specimen
Begin observation
11
9
11
5
Revolving nosepiece
Insert the objective into the light path
and focus the specimen.
5
Fluorescence main switch
Adjust light intensity 11
Coarse and fine focus knobs

Page 19
11. Use of the microscope in Fluorescence (reected light)
11.1 LED switch on
1. Turn the main switch ①. (Fig. 32)
2. Adjust to the desired brightness by rotating the
dial ①.
11.2 Use of uorescence
The lter turret is provided with 4 positions.
• Position “1” is free and is used for Brighteld
• Position “2” contains B (Blue) lter
• Position “3” contains G (Green) lter
• Position “4” is free and is used for Brighteld
• It is not possible to install additional uores-
cence lters.
1. Move the lter selector ②in the desired position.
(Fig. 33)
FILTER
NAME EXCITATION FILTER DICHROIC MIRROR BARRIER FILTER APPLICATIONS
B 450-490 nm 495 nm 520LP nm • FITC: fluorescent antibodies • Achridine orange: DNA, RNA
• Auramine
G500-540 nm 560 nm 575LP nm • Rhodamine, TRITC: fluorescent antibodies • Propidium Iodide: DNA, RNA
• RFP
FFig. 32ig. 32
FFig. 33ig. 33
①
②

Page 20
11.3 Use of light excluding plate
• Microscope is provided with a light excluding
plate that can be placed on the stage and
prevents are and reections coming from
the condenser front lens.
The plate can be used in two different ways.
1. Mode n° 1: place the plate on the stage (under
the slide holder) and place the slide directly over
the plate. (Fig. 34)
2. Mode n° 2: lower the condenser and insert the
plate between the two layers of the stage. (Fig.
35).
• In both cases it is possible to move the sample
using the stage X-Y translation knobs.
FFig. 34ig. 34
FFig. 35ig. 35