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GE InCell Analyzer 6000 High content microscope
JA Bombardier Building, Room 3129
Instrument awarded to Dr. Steve Michnick by the Canadian Foundation for Innovation (CFI)
Advanced Microscope Tier 2 usage price
- Applications
- Transmitted light
- Pseudo phase contrast and DIC
- Fluorescence
- High-throughput imaging
- Long-term imaging
Light sources
LED for transmitted light
Toptica iChrome MLE for fluorescence
Source | Polychroic | Excitation wavelength (nm) | Compatible fluorophores | Nominal Power (mW) |
---|---|---|---|---|
405nm | 390/40 | [370-410] | DAPI, Hoechst | 50 |
488nm | 482/18 | [473-491] | FITC, GFP, YFP | 25 |
561nm | 564/9 | [559-569] | Cy3, DsRed, TxRed | 30 |
642nm | 640/14 | [632-647] | Cy5, Cy5.5 | 50 |
Objectives
- 10x/0.45 Air WD 4.0
- 20x/0.75 Air WD 1.0
- 40x/0.6 Air WD 2.7-3.7
- 60x/0.95 Air WD 0.15
Position | Name | Brand | Full name | Identifier | Workinf distance (mm) | Transmittance (% [nm]) | Technic | Coverslip thickness (mm) |
---|---|---|---|---|---|---|---|---|
1 | 10x/0.45 Air | Nikon | 10x/0.45 Air | MRD00101 | 4.0 | >80% [TBD-TBD] | BF, p-PhC, p-DIC, Fluo | 0.17 |
2 | 20x/0.75 Air | Nikon | 20x/0.75 Air Plan Apo DIC N2 M25x0.75 | MRD00201 | 1.0 | >80% [TBD-TBD] | BF, p-PhC, p-DIC, Fluo | 0.17 |
3 | 40x/0.6 Air | Nikon | 40x/0.6 Air Plan Fluor ELWD M25x0.75 | MRH08430 | 2.7-3.7 | >80% [TBD-TBD] | BF, p-PhC, p-DIC, Fluo | 0.17 |
4 | 60x/0.95 Air | Nikon | 60x/0.95 Air Plan Apo M25x0.75 | MRD00600 | 0.15 | >80% [TBD-TBD] | BF, p-PhC, p-DIC, Fluo | 0.17 |
BF: Bright-field
p-PhC: Pseudo-phase contrast
p-DIC: Pseudo-Differetial interference contrast
- Filters
DAPI
GFP
Cy3
Cy5
- Cy5.5
Position | Name | Brand | Identifier | Polychroic (Transmission) | Emission filter | Efective bandwith (nm) |
---|---|---|---|---|---|---|
1 | DAPI | TBD | TBD | BP 420/20 [430-462] | 455/50 [430-480] | [430-480] |
2 | GFP | TBD | TBD | BP 446/32; 524/42; 600/36; 732/137 | 525/20 [515-535] | [515-535] |
3 | Cy3 | TBD | TBD | BP 446/32; 524/42; 600/36; 732/137 | 605/52 [579-631] | [582-618] |
4 | Cy5 | TBD | TBD | BP 446/32; 524/42; 600/36; 732/137 | 707/72 [671-743] | [671-743] |
5 | Cy5.5 | TBD | TBD | BP 446/32; 524/42; 600/36; 732/137 | 720/60 [690-750] | [690-750] |
- Detector
- sCMOS Monochrome 2560 x 2160 pixels, 16-bit, pixel 6.5um x 6.5um, sensor 16.6mm x 14.0mm
If necessary, turn on the microscope (#1)
Note
The switch is not easily accessible and is located on the back of the right side of the device.
- If necessary, turn on the computer (#2)
- Use your UdeM credentials to log in to Windows
Start the software IN Cell Analyzer 6000
Within the software IN Cell Analyzer:
- Click Eject to open the access door
- Insert your sample in the space provided for this purpose, respecting the orientation indicated
Click Load to close the access doors
First use
During the training you will follow the First use protocol
Important
For use with the 60x objective it is absolutely necessary to use a multi-well plate with a glass bottom. The glass thickness should be 0.17mm. We recommend the following plates:
- Collect your samples
- Click Load to close the access door
- Close the INCell Analyzer software
- Transfer your data to disk D: (Data Storage) or your external hard drive and delete it from local disk C:
- Turn off the computer
Important reminders
- Collect your samples, especially those in the microscope
- Leave the microscope and workspace clean
- Files can be saved temporarily (during acquisition) to local disk C: (desktop)
- At the end of each session, copy your data to your external drive and delete it from local drive C:
- You can store your files on disk D: (Data Storage). If using this disk, please create one folder per laboratory using the principal investigator's last name. Inside, create a folder per user (First Name_Last Name).
Important
In any case, do not store your files on the C: drive.
When using for the first time, it is necessary to define the type of plate used.
Create a new plate
In the INCell Analyzer software:
- Select from the menu Application>Plate\Slide Manager...
- Click on the New Plate\Slide icon
- Select the number of wells in your plate (6, 24, 96 etc...)
- Adjust the following settings:
- Name: Your-Name_Your-Plate
- Plate height, bottom thickness and well volume usually provided in the product technical sheet by the manufacturer
- The material used plastic or glass
- If necessary adjust the number of rows and columns as well as the shape of the well (round or rectangular)
- Click OK
- Close the Plate/Slide Manager window
Measure the bottom height and thickness of the plate bottom
In the INCell Analyzer software:
- Click on the Dashboard
- In Objective Lens select the 10x objective
- On the plate plan, click on the center of a well containing a sample to center this well on the objective
- In the Dashboard menu select Plate/Slide
- Click Verify LAF
- If the 2 detected peaks (blue vertical lines) correspond to the measured peaks (black curve)
- Click Apply Measured Parameters
- Click OK
- Click Yes
- If the 2 detected peaks (blue vertical lines) do not correspond to the measured peaks (black curve)
- Change the bottom thickness value and repeat the operation
- Close the Laser Autofocus Plate/Slide Verification window
Measure A1 well offset
In the INCell Analyzer software:
- if necessary, click on the Dashboard > Objective Lens select the 10x objective
- In the channel list, click the + button and add a transmitted light channel (brightfield)
- To the right of the plate layout, click the Setup Preview Imaging button
- Draw a rectangle around well A1
- Click on preview (on the acquisition panel at the top right of the screen)
- Wait for the preview to be generated
- On the plate map, click on the real center of well A1 to center this well on the objective
In the Dashboard menu select Plate/Slide
Click Edit Plate then Define Upper Left Well
Click Yes
- Check that the yellow circle matches the edges of the well
Click Yes
Compute the inter-well distance
Repeat the offset measurement steps for the bottom right well
- The inter-well distance will then be automatically calculated
The following diagrams allow you to follow the light path in transmitted light (bright field) and in reflected light (fluorescence). These diagrams will be available soon.
- Added to wiki
Troubleshooting
Usually, this microscope displays a green light when operational and an orange light when in use. A red light is on means the microscope is not functional. In this case please contact the platform manager.
FAQ
Can I use this microscope to look at cell in a dish?
- No. This a microscope designed to acquire images of specimen in multi-well plates
- You may also image specimen mounted between a slide and a 0.17mm thick coverslip
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