Olympus Time-lapse Microscope Basic operation

Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
Olympus Time-lapse Microscope
Basic operation
To start up the microscope
1. Switch on the Olympus UCB. (label as
1
)
Power Switch
2. Switch on the MT10. (label as
2
)
Power Switch
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
3. Switch on the camera. (label as
Version 1.0
3
)
Power Switch
4. Turn on the Computer. (label as
4
)
5. Start “Xcellence Pro” in the computer.
(Continue on next page)
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
6. After started the “Xcellence Pro”, you will see the control panel.
4
3
2
1
1
5
4. Microscope
control
3. Camera control
1. Image Buffer
2. Viewport for viewing image
or live view from camera
5. illumination
system control
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Faculty Core Facility
LKS Faculty of Medicine
Olympus Time-lapse Microscope SOP
Version 1.0
Introduction of the Xcellence Pro control panel
In “3. Camera control” panel, you can control the camera and capture image.
Button for
capture an image
Button for
switch on Live
View
Binning selection
Exposure time switch
Brightness adjustment
-Define the maximum
and minimum brightness
value to show in image
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Faculty Core Facility
LKS Faculty of Medicine
Olympus Time-lapse Microscope SOP
Version 1.0
In “4. Microscope control” panel, you can control the microscope.
Objectives Switch
- Switch different objective
Condenser Switch
- Switch different condenser
Lamp Switch
- Switch to turn On/Off lamp and changing intensity
Lamp shutter Switch
- Turn On/Off shutter to stop illumination
Eyepiece & Camera Switch
- Toggle between the microscope
eyepiece and camera
Imaging filter Switch
- For user switch to different filter set
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
In “5. Illumination system control” panel.
Burner Switch
– Start the “Burner” if you need to capture
fluorescence image
Shutter Switch
- Stop the excitation for a short period to avoid
Bleaching of your sample
Intensity Switch
- Adjust the burner intensity level
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Faculty Core Facility
LKS Faculty of Medicine
Olympus Time-lapse Microscope SOP
Version 1.0
To capture fluorescence multi-channel image
1. Switch on the “burner” in “5. Illumination system control” panel if you need to capture
fluorescence image.
2. Switch on the “lamp” in “4. Microscope control” if you need to capture DIC image.
3. Switch the microscope view to “camera” in “4. Microscope control”.
4. Select “live view” in “3. Camera control”.
5. Select the “image filter” you need to use for your image in “4. Microscope control”.
6. Select the objective you want and get the correct focus of your image.
To focus your image, use the focus knob at the microscope stand.
Button to lower
the objective, use
when switching
between 20X &
40X objective
Focus knob
Button to switch between
Coarse/Fine modes
7. Click “Capture” to get your image, the captured image will store at “image buffer”.
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
8. Repeat Step 4 to Step 7 to capture another image.
9. To merge the images you have captured, select one of the images you want to merge in
“image buffer”, then select “Image  combine”. Select the image(s) you need to merge
and then click OK. The merged image will be stored at “image buffer”.
10. To save your image(s) store at image buffer, select the image want to save and then choose
“File Save as” to save your image.
(Please save your file at “D:/User/your name”)
To use experiment manager to capture fluorescence multi-channel,
time lapse and Z-stack image
1. Open your database by select “Database  Open”.
(If you do not have a database, select “Database  Administration  New database”.
In the New database window, enter your name and save your database in
“D:/User/your name”, Then press “Next” followed by “Finish”)
Enter your name
Save in D:/User/your name
2. Open the “Experiment Manager”.
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
3. After select “Experiment Manager”, the “Experiment Manager Panel” is open.
1. Command box
2. Execution Control
4. Experiment
Setting
3. Drawing Bracket
1. Command Box
Image Capture
Multi-color
Time-lapse
Z-stack
Auto-focus
2. Execution Control
Verification
Preparation
Run
Pause
Stop
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
4. To design the image experiment, drag the “command box” into the “3. Drawing bracket”.
4a/ For multi-color imaging
i.
Click and drag
drag 4
ii.
into drawing bracket. E.g. If you need to have 4 color images,
into the drawing bracket. Connect the command with arrow(s).
Set the image capture value in the experiment setting for each command.
You can use “Get Current
Device setting” to get the
current capture value
iii.
Click and drag the
multi-color frame into the drawing bracket, the frame
should include all the capture command boxes.
Multi-color frame
should include all
the command boxes
Arrows connect the
capture command boxes
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
4b/ For Time-lapse imaging
(For time-lapse imaging, you may need to use the incubator, the steps to install the
incubator at page 14)
i.
Click and drag
into drawing bracket. Change the setting at the “4.experiment
setting”.
ii.
Click and drag the
time lapse frame into the drawing bracket, the frame should
include all the capture command boxes.
iii.
Click the time lapse frame; change the setting in the “4.experiment setting”.
iv.
User can also use
auto-focus in time-lapse capture. Click and drag
into the
drawing bracket. Connect the command with arrow(s).
(The auto-focus only available for 20X and 40X objective only. Also ensure you have pushed
the ZDC stick at the microscope stand to “in” position.)
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
4c/ For Z-stack imaging
i.
Click and drag
ii.
Click and drag the
into drawing bracket. Change the setting the experiment setting.
Z-stack frame into the drawing bracket, the frame should
include all the capture command boxes.
iii.
Click the Z-stack frame; Change the setting in the “4.experiment setting”.
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Faculty Core Facility
LKS Faculty of Medicine
Olympus Time-lapse Microscope SOP
Version 1.0
User can also make different combination of experiment command they need.
5. After drawing the experiment plan, select
to verify the experiment plan. If the plan
has error, message will be display. Please modify the plan and verify again.
6. If the plan does not have any problem, select
to allow the system to prepare the
experiment.
7. Select
to run the experiment plan after preparation is complete.
When the experiment plan is running, you can “
Pause” or “
Stop” the plan any
time.
8. After the experiment plan finish, you image file will be store at the “1.image buffer”.
To save your image at image buffer, select the image want to save and then choose “File
Save as” to save your image.
For time-lapse video, you can save the video by choosing “File  Export to AVI”.
(Please save your file at “D:/User/your name”)
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Faculty Core Facility
LKS Faculty of Medicine
Olympus Time-lapse Microscope SOP
Version 1.0
To install the incubator for time-lapse imaging
1. Raise the microscope arm gently.
2. Remove the metal plate from the stage.
3. Remove the slide clips from the stage.
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Faculty Core Facility
LKS Faculty of Medicine
Olympus Time-lapse Microscope SOP
Version 1.0
4. Get the incubator from the plastic tray (Label as A), put it on the stand. It should fit into the
hole on the stage.
5. Use the key to install the screws in order to fix the incubator on the stage.
6. Add 50mL autoclaved water into the incubator with the falcon tube carefully.
7. Switch on the heater. (Label as B) Do not change any setting of the heater.
Power Switch
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
8. Turn on the CO2 mixture gas under the table. The gas flow pressure should set at 15 psi.
DO NOT OVER 15 psi
Meter showing gas level
Turn On
Main switch
Turn on
Regulator
switch
9. Place your culture dish into the incubator. 30 minutes is needed to allow the temperature
reaching equilibrium.
10. Lower the microscope arm gently.
To remove the incubator after time-lapse imaging
1. Switch off the heater. (Label as B) Do not change any setting of the heater.
2. Turn off the CO2 mixture gas under the table.
DO NOT OVER 15 psi
Meter showing gas level
Turn Off
Main switch
Turn off
Regulator
switch
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Faculty Core Facility
LKS Faculty of Medicine
Olympus Time-lapse Microscope SOP
Version 1.0
3. Use the key to remove the screws in order to un-mount the incubator on the stage.
4. Remove the water from the incubator carefully. Use the plastic container to collect the
water. Use paper towel to dry the incubator.
5. Put the incubator back to the tray (Label as A).
6. Install the metal plate and slide clips on to the stage.
7. Lower the microscope arm gently.
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Olympus Time-lapse Microscope SOP
Faculty Core Facility
LKS Faculty of Medicine
Version 1.0
To turning off the microscope
1. Ensure you have saved all the images in the image buffer.
All images in the buffer will be deleted after the Xcellence program is closed.
2. Turn off the burner before closing the Xcellence program. (Important!)
3. Close the Xcellence program.
4. Shut down the computer.
5. Switch off the Olympus UCB. (label as
1
)
2 )
Switch off the camera. (label as 3 )
6. Switch off the MT10. (label as
7.
8. Sign the log sheet before you leave.
End
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