Optika Italy IM-5FLD User manual

Version: 1
Issued: 22, 03, 2018
Model
IM-5FLD
IM-5 Series
INSTRUCTION MANUAL

Page 2
Table of Contents
1. Warning
2. Symbols and conventions
3. Safety Information
4. Intended use
5. Overview
6. Unpacking
7. Assembling
8. Using the microscope
9. Transmitted Light Observation modes
10. Fluorescence Observation mode
11. Simultaneous Phase Contrast + Fluorescence Observation
12. Maintenance
13. Troubleshooting
Equipment disposal

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 that are used in this manual.
CAUTION
This symbol indicates a potential risk and alerts you to proceed with caution.
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 o󰀨 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
For research and teaching use only. Not intended for any animal or human therapeutic or diagnostic use.

Page 4
5. Overview
FIELD STOP
LEVER
PHASE
CONTRAST SLIDER
TRINOCULAR
VIEWING TUBE
(FIXED)
DIOPTRIC
ADJUSTMENT
RING
EYEPIECE
LED
HOUSING
CONDENSER
COARSE
FOCUS KNOB
FINE FOCUS KNOB
TENSION
ADJUSTMENT
COLLAR
OBJECTIVE
STAGE
MICROSCOPE
BODY
LIGHT PATH
SELECTOR
LEVER
STAGE PLATE
LED CENTERING
KNOBS
X-Y STAGE
MOVEMENT
KNOBS
ECO BUTTON
TRANSMITTED
LIGHT INTENSITY
INDICATOR
CONDENSER
CENTERING
SCREWS
APERTURE
STOP LEVER
BRIGHTNESS
ADJUSTMENT
KNOB
(TRANSMITTED
LIGHT)
MAIN
SWITCH

Page 5
5. Overview (opposite side)
FLUORESCENCE LED
LAMP HOUSING
FLUORESCENCE
FILTER SELECTOR
REFLECTED LIGHT
DIAL
FLUORESCENCE
UV PROTECTION
SHIELD
FLUORESCENCE
FILTER INDICATOR
REFLECTED
LIGHT INTENSITY
INDICATOR
DIFFUSION
FILTER

Page 6
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.
7. Assembling
Once you open the box, these are the microscope’s components:
① Microscope body
② Condenser
③ Stage extension
④ Mechanical stage
⑤ Phase contrast slider
⑥ Darkeld slider
⑦ Eyepieces
⑧ Centering telescope
⑨ Metal insert for stage
⑩ Glass insert for stage
⑪ Centering screws
⑫ Power supply
⑬ Power cord
⑭ Green lter (IF550)
⑮ Blue lter (LBD)
⑯ Dust cover
⑰ Holder for Petri diam. 38 mm
⑱ Holder for Terasaki and Petri diam. 65 mm
⑲ Holder for slide and Petri diam. 54 mm
⑳ Lens cleaning tissues
㉑Allen screwdriver
㉒UV protective shield
㉓ Fluorescence lter selector
㉔ Diffusion lter
㉕Lamp housing
⑪
⑩③
⑤
⑥
⑨
①
②
④
⑦
⑧
⑫
⑬
⑭
⑮
⑯
⑰
⑱⑲
⑳㉑㉒
㉓
㉔
㉕

Page 7
③
Fig.2
Fig.3
Fig.4
Fig.5
Fig.1
1. Turn the coarse focusing knob ① until the
nosepiece reaches its lowest position.
► For a safe transport, the nosepiece is
placed in the lowest position and the
tension adjustment collar ②is adjust-
ed to the appropriate tension when the
microscope leaves the factory. (Fig.1)
2. Screw the lowest magnication objective
on to the nosepiece from the right side,
then turn the turret clockwise. Install the
other objectives in the same way, following
the sequence from low to high magnica-
tion.
► Note: the objectives can also be in-
stalled through the stage opening.
(Fig.2)
► Clean the objectives regularly. In inverted
microscopes, the objectives are very sen-
sitive to dust.
► To prevent dust and contamination from
entering the microscope, cover all the un-
used holes with dust caps ③. (Fig.3)
► When operating, use the low magnication
objective (4x or 10X) to search and focus
the specimen, then switch to higher magni-
cations.
► When switching between objectives, slowly
turn the nosepiece until it clicks. The click
means that the objective is in the right posi-
tion, in the center of the light path.
①
②
7.1 Installing the objectives
The stage extension can be installed on either
side of the stage to enlarge the working sur-
face. The mechanical stage must be installed
on the right side of the microscope.
1. Installing the stage extension: screw the
bolts on to the extension, then mount the
extension from below the stage. (Fig.4)
2. Installing the mechanical stage: as for the
extension, the mechanical stage is xed
with two bolts under the stage. (Fig.5)
7.2 Installing the stage extension
and the mechanical stage

Page 8
1. When using the stage inserts, make sure
that the insert is horizontal.
2. Install the stage insert in the stage open-
ing. (Fig.6)
7.3 Installing the stage insert
7.4 Installing the eyepieces
Insert both eyepieces into the tubes of the
optical head. (Fig.7)
Fig.6
Fig.7
Fig.8
1. Slide the condenser into the condenser
holder, inserting the pin on the rear side of
the condenser with the alignment groove
in the condenser holder. (Fig. 8)
2. Lock the condenser with the locking knob
① on the right side of the condenser hol-
der. (Fig. 9)
7.5 Installing the condenser
Fig.9
①
1. Turn the main switch ① to “O”(o󰀨) before
connecting the power cord. (Fig.10)
2. Insert the plug of the provided power
supply into the power socket of the
microscope. (Fig.11)
3. Plug the power cord int the socket of the
power supply (Fig.12)
4. Plug the power cord into the mains socket.
Check for a safe connection.
► Please use the supplied power cord.
If lost or damaged, please refer to
qualied service.
► Connect the power cord to a grounded
power supply only.
7.6 Connecting the power cord
Fig.10
Fig.11 Fig.12
①

Page 9
1. Unscrew the lamp housing lock (Fig. 13)
2. Insert the lamp housing in the
uorescence illuminator connector in the
back side of the microscope. (Fig.14)
3. Tighten the locking screw using the
provided allen screwdriver. (Fig.15)
4. Connect the cable of the lamp hoising
to the connector in the back side of the
microscope. (Fig.16)
7.7 Installing the lamp housing
Fig.13
Fig.14
Fig.15
Fig.16

Page 10
Turning on the light source
Connect the power, turn on the main switch ①.
(Fig.17)
Adjusting the brightness
Turn the brightness adjustment knob ②to
increase and decrease the brightness. (Fig.18)
While rotating the brightness adjustment knob,
the LED indicator on the front panel ③ will turn
ON or OFF some segments. (Fig.19)
Adjusting the tension
► The coarse focusing knob ①is pre
adjusted to a tight tension upon
leaving the factory.
If the nosepiece drops down by itself, or the
specimen defocuses while adjusting the ne
focus knob ③, the coarse focus knob ①is too
loose. Turning the tension adjustment collar
② in clockwise direction tightens the coarse
focus tension. Rotate in the opposite direction
to decrease the tension. (Fig.20)
Setting the specimen
► For the best image quality, use asks,
Petri dishes and slides with a 1.2 mm
thickness.
1. Place the proper insert for your specimen
(according to the table on the right) on the
stage, and x it with the stage clip.
2. Turning the X and Y knobs, move the
specimen to the required position. (Move-
ment Range: 120 (width) × 78 (length)
mm).
Moving the specimen
Move the specimen to the desired position by
freehand or by turning the knobs of the me-
chanical stage.
► When switching objectives, take care
not to touch the adaptor plates with
the objectives, as their weight may
damage the front lens.
8.1 Initial setup
8.2 Stage
8. Using the microscope
Fig.17
Fig.18
①
②
①
②
③
Fig.20
Fig.19
Holder for Petri diameter 38 mm (holder for Terasaki
needed) (provided with the microscope)
Holder for Terasaki and Petri diameter 65 mm
(provided with the microscope)
Holder for slide and Petri diameter 54 mm
(provided with the microscope)
M-793.4
Holder for 2+2 slides.
M-793.6
Holder for Utermöhl-Chamber (holder for Petri 54mm
needed)
Load-bearing side extension
(provided with the microscope)
③

Page 11
Dioptric adjustment
1. Put the scale of each eyepiece on the “0”
position. (Fig 21).
2. Using focusing knobs, focus the specimen
with the right eye.
3. Look into the left eyepiece with your left
eye only. If the image is not sharp, use the
dioptric adjustment ring ①to compensate.
► The adjustment range is ±5 diopter. The
number indicated on the adjustment
ring graduation should correspond to
the operator’s dioptric correction.
Adjusting the interpupillary distance
Observing with both eyes, hold the two
eyepiece prism assemblies. Rotate them
around their common 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. 22)
The range of the interpupillary distance is 48-
75mm.
Selecting the light path
Pull the light path selector lever ③ sideways
using your thumb, selecting the light path you
need. (Fig.23)
8.3 Observation head
②
Fig.21
Fig.22
Fig.23
LIGHT PATH SELECTOR
LEVER
BRIGHTNESS APPLICATION
In 100% used for video or photography Television, and micrography or video
observation
Out 100% used for binocular
observation
Binocular
observation
③
①

Page 12
Using color lters
Selecting the appropriate color lters according
your need. (Fig.24)
You can stack a group of color lters in the lter
holder, if you ensure that they are level and
that the whole thickness is less than 11mm.
Tilting the condenser holder
When a bigger space is needed between
condenser front lens and stage surface, it is
convenient to tilt the condenser. Doing this it is
possible to use the ambient light to illuminate
the specimen. (Fig. 25)
Rotating the illumination pillar
When using big bottles, the illumination pillar
can be rotated out of the light path allowing to
place them on the stage.
Rotate the knob on the right side of the
illumination pillar (Fig. 26) to unlock the
rotation, then swing out the condenser holder
from the light path (Fig. 27).
Once terminated the observation return the
illumination column to its original position and
lock the knob.
8.4 Illumination unit
Fig.24
COLOR
FILTER
USE
Green (IF550) Single contrast color lter used for phase
contrast microscopy
Blue (LBD) Daylight filter
Fig.25
Fig.26
Fig.27

Page 13
Centering the LED
► LED light source is pre centered before
the shipment from the factory. If a new
adjustment is needed, use the three
adjustment knobs ① on the righ side
of the illumination pillar (Fig. 28)
1. Press the ECO button (Fig. 29) to activate
the “ECO” function. Once activated
the system will automatically shut-o󰀨
after 20 minutes from the activation.
2. To deactivate the function, press the
button again.
Fig.28
①
8.5 Eco button
Fig.29
1. Slide the phase slider to the “SL” position
① (Fig. 30).
2. Engage the 10x objective by rotating the
revolving nosepiece, place the specimen
on the stage and adjust approximate
focusing.
3. Use the FS selector lever ② (Fig. 31) to
reduce the eld diaphragm a little.
4. Focus the eld diaphragm by rotating
the height adjustment knob ③ (Fig. 32),
until the edges of the eld diaphragm a re
sharp.
5. Rotate the two centering screws ④on
the condenser so that the eld iris image
becomes concentric with the eld of view
(Fig. 32-33).
6. Using the FS lever ②, open the eld
diaphragm until the eld iris image
inscribes the eld of view. If the image is
found to be eccentric, adjust the centering
again (Fig.31).
7. Open the eld diaphragm so that its image
is almost the same size of (i.e. subscribes)
the eld of view.
8.6 Centering the Field
Diaphragm (FS)
Fig.31
Fig.30
①
②
Fig.32
③
④
FIELD DIAPHRAGM IMAGE
FIELD OF VIEW OF EYEPIECE Fig.33

Page 14
30%
70%
APERTURE DIAPHRAGM IMAGE Fig.37
8.9 Using the Aperture Diaphragm
(Fig. 32)
● In transmitted light brighteld
observation.
The eld diaphragm adjusts the illuminated
area to obtain an image with high contrast.
According to the objective in use, adjust the
FS lever until the iris image circumscribes
the eld of view to block unnecessary light.
●In transmitted light phase contrast
observation
The eld diaphragm must be fully open.
8.8 Using the Field Diaphragm
(Fig. 35)
Fig.36
1. Close the Aperture diaphragm using the
“AS” lever in front of the condenser ①
(Fig. 36-37)
● In transmitted light brighteld
observation, optimum observation is
generally possible by setting the aperture
to between 70% and 80% of the numerical
aperture of the objective.
●In transmitted light phase contrast, the
aperture must be fully opened by operating
on the AS lever.
Fig.35
Fig.34
①
①
When using 4x objective, an additional
lens is needed.
1. Insert the lens ① when 4x objective is in
the light path, remove it when all the other
objectives are used.
If the di󰀨usion lens is used with other
objectives then 4x, the eld diaphragm
wil be not visible any longer.
8.7 Using di󰀨usion lens (Fig. 34)

Page 15
Phase contrast slider
Adjustable phase slider.
● The light ring is pre-centered when the
microscope leaves the factory. It should
therefore need no further adjustment. If
a recentering is needed, it can be per-
formed via the two side bolts.
● The 4x/10x position ① must be used with
4x and 10x phase contrast objectives, the
20x/40x position ② with the 20x and 40x
and the SL position ③ is used for brigh-
eld. (Fig.38)
Installing the phase contrast slider
1. Insert the slider into the condenser, printed
face up.
2. Pull the slider into the desired position, to
the click stop.
3. When in phase contrast observation, keep
the aperture diaphragm adjustment lever
on the “O” (open) position. (Fig.39)
8.10 Phase contrast
Fig.38
Fig.39
① ② ③
SLIDER POSITION MEANING APPLICATION
SL empty hole brighteld observation
4x/10x phase ring 4x/10x phase contrast observation with 4x and
10x objectives
20x/40x phase ring 20x/40x phase contrast observation with 20x and
40x objectives
Darkeld slider (Fig. 40)
Installing the darkeld slider
1. Insert the slider into the condenser, printed
face up.
2. Move the slider into position “DF” ②to
perform darkeld observation.
3. Move the slider into empty position ③to
perform brighteld observation.
4. When in darkeld observation, keep the
aperture diaphragm adjustment lever on
the “O” (open) position.
● Darkeld observation is possible with
objectives having N.A. up to 0.40.
● With higher N.A. the darkeld e󰀨ect
cannot be achieved because of the
condenser N.A.
8.11 Darkeld
Fig.40
②
③

Page 16
Fig.41
8.12 Centering the phase ring
► Usually this operation is not needed.
If necessary, please proceed with the
following steps:
1. Place a specimen on the stage and focus
it.
2. Remove the eyepiece from the sleeve,
and replace it with the centering telescope
(CT). (Fig.41)
3. Check that the phase ring and the objective
match, and that both are steadily set on a
click stop.
4. Use the CT to focus on the phase annulus’s
image ① and the phase contrast ring’s
image ②. If the phase annulus’s image is
not sharp, adjust the CT until you can see a
clear image of the phase annulus. (Fig. 42)
5. Adjust the bolts of the two centering
holes in the phase contrast slider using
the centering screws ③ until the phase
annulus center and the phase contrast ring
center coincide. (Fig. 43)
6. The 4x and the 10x phase contrast
objectives use the same ring on the
phase contrast slider. The coincidence of
the phase annulus center and the phase
contrast center must be veried with both
objectives.
► If the phase annulus is incorrectly
centered, the contrast will be severely
impaired.
► The phase ring may need recentering
during and after observation of very
thick specimens.
► The phase ring may show an apparent
misalignment if the cover glass is not
at. Fig.43
①
②
Fig.42
③

Page 17
Fig.44
8.13 Fluorescence
► Fluorescence illuminator has a
dedicated LED light source.
► Insertion of dedicated LED is motorised:
the LED switching is automatical when
moving the ltercube selector.
► If the reected light dial is in “O” (OFF)
position, the LED will not move when
the ltercube selector is moved.
1. Switch on the uorescence illuminator by
turning the reected light intensity dial ①.
The LED scale for the reected ligth ②
and the lter cube indicator ③ will turn on
(Fig.44).
2. Move the ltercube selector ④ in one of
the 4 position available until it moves to a
click stop. (Fig.45a, b, c, d). The lter cube
indicator ③ will change accordingly (Fig.
46).
► If the ltercube selector is not moved
in a precise click stop, the LED will
not change and the ltercube indicator
panel will not light up.
► The ltercube selector has 4 positions
marked with colored rings. The
correspondence of the ltercube
selector and the the ltercube indicator
panel is the following:
Colored ring
(figure)
Letter on
the panel
Filter
BLUE (45a)BBlue
GREEN (45b) G Green
WHITE (45c)UV Ultra violet
WHITE (45d)OP EMPTY
3. Place a uorescent specimen on the stage.
4. Insert the desired objective in the light path
and focus it.
5. Change the light intensity using the light
intensity dial ①according to the specimen. Fig.46
Fig.45a
Fig.45c
Fig.45b
Fig.45d
FILTER
CUBE
EXCITATION
FILTER
DICHROIC
MIRROR
EMISSION
FILTER
APPLICATION
UV 365/50 nm 400 nm 420LP nm
• Autofluorescence
• DAPI: staining for DNA
• Hoechst: chromosomes
B470/40 nm 495 nm 525/50 nm • FITC: fluorescent antibodies
• Achridine orange: DNA, RNA
• Auramine
G 560/40 nm 585 nm 645/75 nm • Rhodamine, TRITC: fluorescent antibodies
• Propidium Iodide: DNA, RNA
• RFP
①
②
③
④
③

Page 18
Fig.47
8.14 Centering the Field Diaphragm
1. Insert the lter B by moving the ltercube
selector ①. (Fig. 49). The “B” LED on the
front panel will light up ④. (Fig.48).
2. Engage the 10x objective by rotating the
revolving nosepiece, place the specimen
on the stage and adjust approximate
focusing.
3. Use the FS selector lever ② (Fig. 47) to
reduce the eld diaphragm a little.
5. Using the provided allen wrench, rotate the
two centering screws ③ so that the eld iris
image becomes concentric with the eld of
view (Fig. 48).
6. Using the FS lever ②, open the eld
diaphragm until the eld iris image inscribes
the eld of view. If the image is found to
be eccentric, adjust the centering again
(Fig.48).
7. Open the eld diaphragm so that its image
is almost the same size of (i.e. subscribes)
the eld of view.
Fig.48
③
● In reected light uorescence
observation.
The eld diaphragm adjusts the illuminated
area to obtain an image with high contrast.
According to the objective in use, adjust the
FS lever until the iris image circumscribes
the eld of view to block unnecessary light.
8.15 Using the Field Diaphragm
①
②
FIELD DIAPHRAGM IMAGE
FIELD OF VIEW OF EYEPIECE Fig.50
④
Fig.49

Page 19
Installing the adapter
1. To activate the video port, pull the light
path selector lever to “In” position. (Fig.51)
2. Loosen the locking bolt ① on the trinocular
viewing tube, and remove the dust cap.
3. Install the C-mount adapter into the
trinocular port according to its instructions,
and screw down the locking bolt ①. (Fig.
52)
4. Attach the camera ② to the C-mount
adapter.
● Warning: for some cameras (mainly
reex) the ring is not included with the
microscope, and it should be supplied by
the user.
● For the photography of dark specimens,
obscure the eyepieces and the viewnder
with a dark cloth in order to reduce stray
light.
● The camera magnication can be
calculated as objective magnication ×
camera + lens magnication.
► When shooting with a SLR, the
mirror movement may cause camera
movement. Please lift the mirror,
use long exposure times and use an
extension cord.
8.16 Microphotography
Fig.51
Fig.52
①
②

Page 20
9. Transmitted Light Observation mode
① Phase contrast slider (pag.13)
(Controls used ) (Pag.)
② Main switch (pag. 8)
④ Light path selector (pag. 16)
⑤ Stage plate (pag. 8)
⑥ X/Y axis knobs (pag. 6)
⑦ Revolving nosepiece (pag. 6)
⑨ Brightness adjust. knob (pag. 8)
⑩ Binocular tube (pag. 10)
⑪ Diopter adjustment ring (pag. 10)
⑫ AS lever (pag. 13)
⑬ FS lever (pag. 13)
⑦ Revolving nosepiece (pag. 6)
⑭ Filter insertion slot (pag. 11)
⑨ Brightness adjust. knob (pag. 8)
⑧ Coarse/ne focussing knobs (pag. 8)
⑧ Coarse/ne focussing knobs (pag. 8)
TRANSMITTED LIGHT BRIGHTFIELD/PHASE CONTRAST OBSERVATION PROCEDURE
The following ow shows the basic operating procedure for transmitted light brighteld or phase contrast observation.
Select the brightfield (BF) or phase contrast (PH).
Set the main switch to “I” (ON).
Select the light path.
Place the specimen on the stage.
Insert 10x objective in the light path.
Bring the specimen in focus.
Begin observation
Adjust aperture diaphragm and field diaphragm.
Open both diaphragms in case of PH observation.
Engage the desired objective in the light path and bring in
focus.
Adjust brightness.
Insert the required filters.
Adjust interpupillary distance.
Adjust the diopter.
Adjust brightness
③ Reected light dial (pag. 16)
Set the reflected light dial to “O” (OFF).