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| title | Description |
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| DescriptionLight sources| Expand |
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| title | Complete specifications |
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| Emission peak (nm) | Nominal Power (mW) | Measured Power (mW)
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| 385/30 | 150 | | | 469/38 | 110 | | | 555/30 | 31 | | | 631/33 | 50 | |
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Objectives| _Product Information.pdf | | height | 250 |
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Image AddedZeiss Colibri 7 Spectra
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Objectives2.5x2.5x/0.075 Air - 10x/0.25 Air Ph1
- 20x/0.5 Air Ph2
40x/0.75 Air Ph2 - 63x/0.75 Air Ph2 Long Distance
- 63x/1.4 Oil
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| title | Complete specifications |
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| | Position | Name | Brand | Full name | ID | Magnification | Numerical Aperture | Immersion | Type | Working distance (mm) | Transmittance (% [nm]) | Technique | Cover glass thickness (mm) |
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| 1 | 2.5x/0.075 Air | Zeiss | 2.5x/0.075 EC Plan-Neofluar | M27M27 | 420941-9911-000 | 10x | 0.25 | Air | N-AchroPlan | 6.5 | Not Available | BF, PhC, Fluo | 0.17 | | 3 | 20x/0.5 Air | Zeiss | 20x/0.50 Ph2 EC Plan-Neofluar | M27420351-9910-000 | 20x | 0.50 | Air | Plan Neofluar | 2.0 | Not Available | BF, PhC, Fluo | 0.17 | | 4 | | Zeiss | 40x/0.75 Ph2 EC Plan-Neofluar | M27 | 420361-9910-000 | 40x | 0.75 | | Plan Neofluar | 0.71 | Not Available | BF, PhC, Fluo | 0.17 | | 5 | 63x/0.75 Air
| Zeiss | 63x/0.75 Corr Ph2 LD Plan-Neofluar | M27| 421381-9970-000 | 63x | 0.75 | Air | LD Plan-Neofluar | 1.7 at cover glass 0.75 | Not Available | BF, PhC, Fluo | 0 - 1.5 | | 6 | 63x/1.4 Oil | Zeiss | 63x/1.4 DIC Plan-Apochromat Oil | M27 |
Filter
- DAPI
- GFP
- Rhodamine
- DHE (dihydroethidium)
- Cy5
- Quadruple DAPI/GFP/Cy3/Cy5
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| title | Complete specifications |
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Detectors- PCO Edge 5.5Zeiss AxioCam MRm
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| title | Complete specifications |
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Zeiss AxioCam MRmCCD| sCMOS | Sensor Category
| Monochrome | Nb Pixels
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14 1388 104045 89 x 67 11 12| 16-bit | | Speed at full resolution |
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13 55 Readout 8 Pelletier| Forced air +7C | Dark Current
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7 | 6 e⁻/pixel/sec | Full well capacity
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17 2000FireWire (IEEE 1394a)| Dual Camera Link PCIe | Mount
| C-mount |
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| name | PCO_Edge 5.5_Datasheet.pdf |
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| height | 250 |
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- Zeiss Axiocam MRm (not installed)
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| title | Complete specifications |
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| | CameraPCO Edge 5.5 | Zeiss Axiocam MRm |
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Sensor Type | sCMOSCCD | Sensor Category
| Monochrome | Nb Pixels
| 51. | 5 4 M | Pixel Layout | 2560 1388 x | 21601040 | Pixel size | 6. | 5 45 um
| Sensor size
| 168.9 mm x 6 | mm x 14. | 0 7 mm | Sensor diameter
| 21.8 11 mm
| Bit depth
| 1612-bit | | Speed at full resolution | 100 13 images/s
| Max QE
| 60 55 % at 600 nm | | Reading Readout noise | 1.0 8 e⁻ | Cooling
| Forced air +7CPelletier | Dark Current
| 0.6 7 e⁻/pixel/sec | Full well capacity
| 30 17 000 e-
| Dynamic Range
| 1:300002000 | Interface
Dual Camera Link PCIe | FireWire (IEEE 1394a) | Mount
| C-mount |
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| auitabspage| View file |
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| name | Zeiss_AxioCam MRm_Datasheet.pdf |
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| height | 250 |
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| id | User Guide |
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| id | User Guide |
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| title | User Guide |
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| User Guide| UI Expand |
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| - If not already done, turn on the the computer (#1) and use your UdeM credentials to log in to Windows using your UdeM credentials
- Remove the dust cover from the microscope
- If incubation is required, turn on the incubation power bar (#2) on the desk near the computer and open the CO₂ cylinder (#2B) near the sink
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Make sure the humidifier is properly filled with distilled water. |
Turn on the microscope power bar bar (#3) on the desk near the computer - When using the instrument for the first time, it is necessary to import the microscope -specific parameters BEFORE configuration before starting the software. See the the First Use section below.
- Start Zen
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| | Please note, When using the instrument for the first time, it is necessary to import the microscope -specific parameters configuration into the software. This procedure is usually carried out during the training session. However, it is can also possible be used to use it to reset the software if it is does not displayed display correctly, for example. | Note |
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Running this procedure will delete erase all your experiment protocols and restore reset the software to its original settings (ask for support if you are not sure).
If Zen is open, close Zenit and wait for until it to close has completely shut down (this may take up to 30 seconds) On the Desktop, open the Softwares folder folder Double-click Zen Settings for Axio-Osberver Zeiss Z1-Colibri A script will run and a black window will appear briefly Click OK to close the message When the message Settings for Zen have been imported successfully . appears, click OK to close it You can then now open Zen Zen
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| During this procedure, you will: This procedure puts in and performs - Perform a calibration
- Load your sample
- Find and adjust the focus
Once completed, your sample .At the end of this procedure the microscope will be ready for acquisition.
| UI Expand |
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| This step is required to calibrate the microscope in XY and Z. Doing a Performing this calibration will save you a lot of time to find your significantly reduce the time needed to locate and focus on your sample. | UI Expand | title | Calibration within the Zen Software | On the microscope touch screen: press Home>Microscope>Control>Objectives>- select the lowest-magnification objective. On the microscope touch screen Home > Microscope > Control > Objectives > 2.5x
to select a safe objective:Once Zen software is popup - calibration dialog should
show up, just - appear. Simply click Calibrate Now.
The microscope will lower the objectives, perform a and then - first, followed by a Z calibration, and then return to its original position.
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Once calibrated, the focus can be is typically found at Z = 1.1 mm for a microscope slide ). The Z value can be found . For a multi-well plate the focus is generally around Z = 3 mm but this is highly dependent on your multi-well plate. The Z position can be viewed on the microscope touch screen Home>Zunder Home > Z-Position If the calibration popup dialog did not show up you at Zen opening, you can do it manually: Select the Stage window (, as well as in Zen within the Focus tab, located on the right of Zen window) and click Calibrate A warning message will show up click Continue The microscope will lower the objectives perform a XY calibration
Then in the Focus Window on the right of Zen window) and click Calibrate A warning message will show up click Continue The microscope will lower the objectives perform a Z calibration Done. nce calibrated, the focus can be found at Z = 1.1 mm for a microscope slide). The Z value can be found on the microscope touch screen Home>Z-Position | | UI Expand |
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| title | Calibration with the touch screen |
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| side of the screen. |
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| title | The calibration dialog did not appear... |
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| The system may already be calibrated. This can occur if a previous user calibrated the system and left it on. On the microscope touch screen: If not already done, Press Home>Load Position to lower the objectives to the lowest positionPress | UI Expand |
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| title | Verifying if the system is already calibrated |
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| In Zen, within the Focus tab, located on the right side of the screen: Click Load to lower the objective - The Z position is indicated below Current
- If Z position value is less than 100 um the system is calibrated
On the microscope touch screen: Lower the objective by pressing Home > Load Position Press
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Set Work Position to store this position If necessary,
| move slightly adjust the focus
| slightly up remove limit reached Limit Reached message displayed on the
| touchscreenIf not already done, press Home>Microscope>Control>Objectives>5x to select the 5x objectiveIf asked, tap Done to remove the oil lens cleaning warningPress Home>Microscope>XYZ>Position>Z-Position>Set zero>Auto to perform focus calibrationPress OK to start the calibration procedureWait a few seconds for the calibration to be completedPress Home>Microscope>XYZ>Position>XY-Position>Set zero>Auto to perform a stage calibrationPress OK to start the calibration procedureWait a few seconds for the calibration to be completed| Note |
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Once calibrated, the focus can be found at Z = 1.7 mm). The Z value can be found on the microscope touch screen Home>Z-Position |
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| UI Expand |
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| touch screen - The Z position value is indicated
- If the value is less than 0.1 mm then the system is calibrated
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| UI Expand |
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| title | Manual calibration with the software |
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| In Zen, within the Stage tab, located on the right side of the screen: - If not already done, check the Show All option
At the bottom of the tab click Calibrate A warning message will show up click Continue The microscope will lower the objectives and perform a XY stage calibration and then return to its original position.
In Zen, within the Focus tab, located on the right side of the screen: - If not already done, check the Show All option
At the bottom of the tab click Calibrate A warning message will show up click Continue The microscope will then perform a Z calibration and then return to its original position
XY and Z calibrations are now complete |
| UI Expand |
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| title | Manual calibration with the microscope touch screen |
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| | Warning |
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| Make sure to calibrate the focus before performing the first focus. | On the microscope touch screen: |
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- If not already done, press Home>Microscope>Turret>Objectives>5x to select the 5x objective
| Info |
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The 5x objective is the safest because it has the longest working distance (12mm). The sample will appear perfectly sharp long before the lens approaches it. It is recommended to always first focus with the safest lens. The objectives are parafocal, focusing with the safest objective will then allow you to easily find your sample with another objective. The 10x objective is also safe because its working distance is 6.5 mm. |
- If not done already, press Home>Load Position to lower the objective to the lowest position and press Set Work Position to store this position
- If necessary, move the focus slightly up to remove the Lower Z limit reached message displayed on the touchscreen
- Place the test slide on the microscope stage with the coverslip toward the objective
| Note |
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| Always use the test slide to perform the first focus. |
- If necessary, move the stage so that the sample is centered on the objective
On the computer: Open Zen In the Locate tab, select BF or the desired fluorescence (DAPI, GFP, mPlum) to activate the configurationAdjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
| Note |
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Once calibrated, the focus can be found at Z = 1.7 mm). The Z value can be found on the microscope touch screen Home>Z-Position | In the Locate tab, select Off to turn off the illuminationNavigate to Home > Microscope > XYZ > Position > Z-Position > Set Zero > Auto to perform focus calibration Press OK to start the calibration procedure Wait a few seconds for the calibration to complete Lower the objective by pressing Home > Load Position Press Set Work Position to store this position If necessary, slightly adjust the focus upward to clear the Lower Z Limit Reached message displayed on the touch screen Navigate to Home > Microscope > XYZ > Position > XY-Position > Set Zero > Auto to perform a stage calibration Press OK to start the calibration procedure Wait a few seconds for the calibration to complete
XY and Z calibrations are now complete |
| UI Expand |
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| title | Ensuring the calibration dialog is displayed at startup |
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| In Zen: In the menu bar, navigate to Tools > Options Select Startup/Shutdown Under Stage/Focus Calibration, ensure Request Stage/Focus Calibration on Startup is checked Click OK to close the Options dialog
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| | Warning |
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| | Ensure that the calibration has been completed beforehand. Calibration will significantly reduce the time required to locate and focus on your sample. |
On the microscope touch screen: - If not already done, select the lowest-magnification objective Home > Microscope > Control > Objectives > 2.5x
| Info |
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The 2.5× and 10× objectives are the safest to use due to their long working distance (>6 mm). The sample will appear in sharp focus well before the objective approaches it. It is recommended to always focus using the safest objectives first. Since the objectives are parafocal, focusing with the safest objectives will facilitate locating the sample when switching to higher-magnification objectives.
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Lower the objective by pressing Home > Load Position Press Set Work Position to store this position If necessary, slightly adjust the focus upward to clear the Lower Z Limit Reached message displayed on the touch screen - Place the test slide on the microscope stage with the coverslip facing the objective
| Note |
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| Using a test slide will significantly reduce the time required to set up the instrument. |
- If necessary, adjust the stage to ensure that the sample is centred under the objective
In Zen: - In the Locate tab, select BF or the desired fluorescence (Fluo, DAPI, GFP, Cy3, Cy5) to activate the configuration
- Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
| Info |
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Once calibrated, the focus is typically found at Z = 1.1 mm for a microscope slide. For a multi-well plate the focus is generally around Z = 3 mm but this is highly dependent on your multi-well plate. The Z position can be viewed on the microscope touch screen under Home > Z-Position, as well as in Zen within the Focus tab, located on the right side of the screen. |
- In the Locate tab, select Off to turn off the illumination
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| UI Expand |
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| | Warning |
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| Perform the initial focus using the safest objective before switching to higher-magnification objectives. |
| UI Expand |
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| title | Focusing with air objectives |
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| After performing the initial |
| | UI Expand |
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| | Warning |
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| First focus with the safest objective before selecting another lens and continuing with secondary focus. |
| UI Expand |
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| title | Focusing with air objectives |
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| After performing the first focus, on the microscope touch screen: Press Home>Microscope>Control>Objectives, press 10x, 20x or 40x to select the desired objective| Info |
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The 40x objective is the best Air objective because it has the greatest number of optical corrections (Plan Apochromat) and the largest numerical aperture (0.95) but has a smaller field of view. The 20x/0.8 objective offers the best compromise between Resolution and Field of View |
| Note |
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There are two (2) 40x objectives, make sure you select the Air 40x |
In Zen : - In the Locate tab, select BF or the desired fluorescence (DAPI, GFP, mPlum) to activate the configuration
- Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
- In the Locate tab, select Off to turn the illumination off
- Your sample is ready for acquisition!
| UI Expand |
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| title | Focusing with oil objectives |
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| After performing the first focus, on the microscope touch screen: - Press
Home>Microscope>Turret>- Home>Microscope>Control>Objectives
- Press
63x Oil- 10x, 20x,
40x Oil - 40x or 63x (0.75) to select the desired objective
. The microscope will automatically lower the stage so that the sample is accessible. and 63x oil objectives provide the same spatial resolution because they have the same numerical aperture (1.4). The 40x oil objective offers a larger field of view and transmits light slightly better beyond 700nm. The 63x oil objective transmits light slightly better in the visible spectrum (440-710 nm) and has a better Strehl ratio (90%). It is particularly suited for super-resolution imaging, but its field of view is smaller. objective is the best Air objective because it has the largest Numerical aperture (0.75) together with a wider field of view (40x).
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In Zen: - In the Locate tab, select BF or the desired fluorescence (Fluo, DAPI, GFP, Cy3, DsRed, Cy5) to activate the configuration
- Adjust the
| Note |
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There are two (2) 40x objectives, make sure you select the 40x Oil | Place a single drop of oil on the objectivePress Done. The microscope will automatically return the objective to its original positionIn Zen : - In the Locate tab, select BF or the desired fluorescence (DAPI, GFP, mPlum) to activate the configuration
Adjust the - focus with the precision dial while looking through the eyepieces
until the - until the image is perfectly sharp
- In the Locate tab,
select - select Off to turn off the illumination
off
Your sample is ready for acquisition! |
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This procedure puts the microscope in a safe configuration and performs a focus calibration. At the end of this procedure the microscope will be ready for acquisition. | UI Expand |
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| | On the microscope |
| Loading samples | | Focusing with oil objectives |
| After performing the initial focus, on the microscope | Home>Load Position to lower the objectives to its lowest positionPress Set Work Position to store this positionIf necessary, move the focus slightly up to remove the Lower Z limit reached message displayed on the touchscreenPress Home>Microscope>Control>Objectives>2.5x to select the 2.5x objectiveIf asked, tap Done to remove the oil lens cleaning warningPress Home>Microscope>XYZ>Position>Z-Position>Set zero>Auto to perform focus calibrationPress OK to start the focus calibration procedureWait a few seconds for the calibration to be completed| Note |
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Once calibrated, the focus can be found at Z = 1.1 mm). The Z value can be found on the microscope touch screen Home>Z-Position |
| | UI Expand |
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| | Warning |
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| Make sure to calibrate the focus before performing the first focus. |
On the microscope touch screen: - Press Home>Microscope>Turret>Objectives
- Press 2.5x to select the 2.5x lens
| Info |
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The 2.5x objective is the safest because it has the longest working distance (9 mm). The sample will appear perfectly sharp long before the lens approaches it. It is recommended to always first focus with the safest lens. The objectives are para-focal, focusing with the safest objective will then allow you to easily find your sample with another objective. |
- Press Home>Load Position to lower the objectives to its lowest position
- Press Set Work Position to store this position
- If necessary, move the focus slightly up to remove the Lower Z limit reached message displayed on the touchscreen
- Place the test slide on the microscope stage with the coverslip toward the objective
| Note |
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| Always use the test slide to perform the first focus. |
- If necessary, move the stage so that the sample is centered on the objective
On the computer: Open ZenIn the Locate tab, select BF or the desired fluorescence (DAPI, GFP, Rhodamine, etc…) to activate the configurationAdjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
| Note |
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Once calibrated, the focus can be found at Z = 1.1mm). The Z value can be found on the microscope touch screen Home>Z-Position | In the Locate tab, select Off to turn off the illumination| UI Expand |
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| | Warning |
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| First focus with the safest lens before selecting another lens and continuing with secondary focus. |
| UI Expand |
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| title | Focusing with air objectives |
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| After performing the first focus, on the microscope touch screen: Press Home>Microscope>Control>ObjectivesPress 20x or 40x or 63x to select the desired lens
| Info |
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The 40x objective is the best Air objective because it has the largest numerical aperture (0.75). | Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharpYour sample is ready for acquisition!| UI Expand |
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| title | Focusing with oil lenses |
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| After performing the first focus, on the microscope touch screen:
- Press Home>Microscope>Control>Objectives
Press 63x Oil. The microscope will automatically lower the objective so that the sample is accessible - Place a drop of oil on your sample
- Press Done. The microscope will automatically return the sample to its original position
In Zen: - In the Locate tab, select BF or the desired fluorescence (DAPI, GFP, Rhodamine, etc…) to activate the configuration
- Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
- In the Locate tab, select Off to turn the illumination off
- Your sample is ready for acquisition!
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| UI Expand |
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| - Files can be saved temporarily (during acquisition) on the local C: drive (desktop)
- At the end of each session, copy your data to your external drive and delete it from the local C: drive
- You can store your files on the D: drive (Data Storage). If you do, please create a folder per laboratory using the principal investigator last name. Within, create one folder per user (Firstname_Lastname).
| Note |
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In any case, your files should be removed from the C: drive. |
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| UI Expand |
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| - Save your data
- Close Zen
- Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
If used, turn off the incubation module power strip (#2A) and close the CO2 cylinder (#2B) Select the 2.5x objective and press load position to bring the objectives to the bottom position
Turn off the microscope power bar (#3) - Turn off the computer
- Cover the instrument with the protective dust cover
| Note |
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| - Take back your samples including ones in the microscope
- Leave the microscope and the working area clean
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| | Tabs Page |
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| id | Lightpath |
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| title | Lightpath |
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| The following schematics depict the light path for transmitted (bright-field and Phase Contrast) and reflected (fluorescence) lights. | View file |
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| name | Zeiss_Z1-Colibri_LightPath.pdf |
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| height | 250 |
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| Available manuals |
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