Axio-Observer | direction | horizontal |
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| id | Description |
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Zeiss AxioObserver Z1 inverted microscopeMicroscope
Desmarais Building, Room 2234
Simple Microscope usage price
Instrument awarded to Dr. Audrey Claing and Dr. Jean-Philippe Gratton by the Canadian Foundation for Innovation (CFI) in 2014
- Inverted microscope
- Widefield imaging
Bright-field- Phase contrast
- Fluorescence
- Incubation
Light sources
LED lamp for transmitted light
- Timelapse imaging
- Deconvolution
- Extended Depth of Focus
Image AddedClick to enlarge
Image AddedClick to enlarge
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| title | Description |
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| DescriptionLight sources |
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Colibri 7 - (385/469/555-590/631) for fluorescence
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Full specifications of the Zeiss Colibri 7 light source | Emission peak (nm) | Nominal Power (mW) | Measured Power (mW)
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| 385/30 | 150 | | | 469/38 | 110 | | | 555/30 | 31 |
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3252 | Information Zeiss Colibri 7
Objectives
20x/0.5 Air Ph2 WD 2.0Empty63x/1.4 Oil DIC WD
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| name | Zeiss_Colibri-7_Product Information.pdf |
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| height | 250 |
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Image AddedZeiss Colibri 7 Spectra
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Objectives2.5x/0.075 Air
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WD 2.0 WD 6.5- 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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Objectives complete | | 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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20x5 20x50 Ph2420351M27991020x | -20800, PhCEmpty | 3 | 63x/1.463x14 PlanApochromat OilM27420782-990063x14OilPlan Apochromat0.19 | >80% [450-750] | BF4 | Empty | 5 | 2.5x075 2.5x075420320-9901M27 2.5x075-95>90% [400-750]| 0 | Not Available | BF, PhC, Fluo | 0.17 |
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610x25 10x25 Ph1N-AchroplanM27 420941-991110x25AchroPlan65>80% [400-800]| 71 | Not Available | BF, PhC, Fluo | 0.17 |
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Filter cubesDAPIGFPDsRedDHE (dihydroethidium)Cy5Quadruple DAPI/GFP/Cy3/Cy5
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| title | Filters complete specifications |
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| Position | Name | Brand | ID | Excitation filter | Dichroic mirror | Emission filter | Comments |
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1 | DAPI Filter Set 49 | Zeiss | 488049-9901 | 365/50 [325-375] | 395LP | 445/50 [420-470] | 2 | GFP Filter Set 13 | Zeiss | 488013-0000 | 470/20 [460-480] | 495LP | 517/25 [505-530] | 3 | DsRed Filter Set 43 | Zeiss | 000000-1114-101 | 545/25 [533-567] | 570LP | 605/70 [570-640] | | 4 | DHE | Custom | Custom | 500/50 [475-525] | 540LP | 580/20 [570-590] | Undefined specifications Best guess values |
5 | Cy5 Filter Set 50 | Zeiss | 488050-9901 | 640/30 [625-655] | 660LP | 690/50 [665-715] | 6 | Quadruple DAPI/GFP/Cy3/Cy5 FS90 HE LED | 489090-9110-000QBS 405 + 493 + 575 + 653 | QBP 425/30+514/30+592/25+709/100 | Excitation filters included in the light source FS90 HE LED | - Detector
- Zeiss AxioCam MR R3 CCD Camera 1388 x 1040 pixels, 12-bit, 13 images/s at full resolution, detector size 8.9 mm x 6.7 mm
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| id | User Guide |
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| title | User Guide |
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Remove the dust cover from the microscope Turn on the computer (#1) - If necessary, turn on the power bar for the incubation module (#2A) and open the CO2 cylinder (#2B)
Turn on the power bar at the left of the computer monitor (#3) - Log in Windows using your UdM credentials
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When using for the first time, it is necessary to import the microscope-specific parameters BEFORE starting the software. See the First Use section below. |
- Start the Zen software
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When using for the first time, it is necessary to import the microscope-specific parameters into the software. This procedure is usually carried out during the training session.However, it is also possible to use it to reset the software if it is not displayed correctly, for example.| 5 | 63x/0.75 Air
| Zeiss | 63x/0.75 Corr Ph2 LD Plan-Neofluar | 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 | 420782-9900-000 | 63x | 1.4 | Oil | Plan Apochromat | 0.19 | >80% [400-700] | BF, Fluo | 0.17 |
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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 5.5
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| title | Complete specifications |
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| | Camera | PCO Edge 5.5 |
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Sensor Type | sCMOS | Sensor Category
| Monochrome | Nb Pixels
| 5.5 M | Pixel Layout | 2560 x 2160 | Pixel size | 6.5 um
| Sensor size
| 16.6 mm x 14.0 mm | Sensor diameter
| 21.8 mm
| Bit depth
| 16-bit | | Speed at full resolution | 100 images/s
| Max QE
| 60 % at 600 nm | | Reading noise | 1.0 e⁻ | Cooling
| Forced air +7C | Dark Current
| 0.6 e⁻/pixel/sec | Full well capacity
| 30 000 e-
| Dynamic Range
| 1:30000 | Interface
| 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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| | Camera | Zeiss Axiocam MRm |
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Sensor Type | CCD | Sensor Category
| Monochrome | Nb Pixels
| 1.4 M | Pixel Layout | 1388 x 1040 | Pixel size | 6.45 um
| Sensor size
| 8.9 mm x 6.7 mm | Sensor diameter
| 11 mm
| Bit depth
| 12-bit | | Speed at full resolution | 13 images/s
| Max QE
| 55 % | | Readout noise | 8 e⁻ | Cooling
| Pelletier | Dark Current
| 0.7 e⁻/pixel/sec | Full well capacity
| 17 000 e-
| Dynamic Range
| 1:2000 | Interface
| FireWire (IEEE 1394a) | Mount
| C-mount |
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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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| title | User Guide |
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| User Guide| UI Expand |
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| - If not already done, turn on the computer (#1) and 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 (#3) on the desk near the computer - When using the instrument for the first time, it is necessary to import the microscope configuration before starting the software. See the First Use section below.
- Start Zen
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| When using the instrument for the first time, it is necessary to import the microscope configuration into the software. This procedure is usually carried out during the training session. However, it can also be used to reset the software if it does not display correctly, for example. Running this procedure will erase all your experiment protocols and reset the software to its original settings (ask for support if you are not sure).
If Zen is open, close it and wait until it has completely shut down (this may take up to 30 seconds) On the Desktop, open the Softwares folder Double-click Zen Settings for Zeiss Z1-Colibri A script will run and a black window will appear briefly When the message Settings for Zen have been imported successfully appears, click OK to close it You can now open Zen
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| During this procedure, you will: - Set the microscope to a safe configuration
- Perform a calibration
- Load your sample
- Find and adjust the focus
Once completed, your sample will be ready for acquisition.
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| This step is required to calibrate the microscope in XY and Z. Performing this calibration will significantly reduce the time needed to locate and focus on your sample. - If not already done, select the lowest-magnification objective. On the microscope touch screen Home > Microscope > Control > Objectives > 2.5x
- In Zen, once it has started a calibration dialog should appear. Simply click Calibrate Now.
The microscope will lower the objectives, perform an XY calibration first, followed by a Z calibration, and then return to its original position.
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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. |
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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. | 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 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 - The Z position value is indicated
- If the value is less than 0.1 mm then the system is calibrated
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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 |
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| title | Manual calibration with the microscope touch screen |
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| On the microscope touch screen: Navigate 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
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Please note, this procedure will delete all your experiment protocols and restore the software to its original settings. |
- If open, close the Zen software and wait for it to close completely (up to 30 seconds)
- On the Desktop open the Documentation folder
- Double-click Settings for Axio-Osberver Z1
- A script will run and a black window will appear briefly
- You can then reopen the Zen software
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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.
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On the microscope touch screen:
Press Home>Load Position to lower the stage to its lowest positionPress Set Work Position to store this position If necessary,
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move slightly adjust the focus
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slightly up remove “Lower limit reached” Limit Reached message displayed on the
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touchscreentouch screen Navigate to Home > Microscope > XYZ > Position > XY-Position > Set Zero > Auto to perform a stage
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Press Home>Microscope>Turret>Objectives>10x to select the 10x objectiveIf asked, tap Done to remove the oil lens cleaning warningPress Home>Microscope>XYZ>Position>Z-Position>Set zero>Auto to perform focus calibration Press OK to start the
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focus calibration procedure Wait a few seconds for
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the calibration to be completed| Note |
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the calibration to complete
XY and Z calibrations are now complete |
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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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Once calibrated, the focus can be found at Z = 1.5 mm). The Z value can be found on the microscope touch screen Home>Z-PositionFirst Make sure to calibrate the focus before performing the first focusPress 10x to select the 10x lens| 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: |
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Press Home>Microscope>Turret>Objectives- If not already done, select the lowest-magnification objective Home > Microscope > Control > Objectives > 2.5x
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10x objective is the safest because it has the longest 2.5× and 10× objectives are the safest to use due to their long working distance ( |
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6.5 >6 mm). The sample will appear |
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perfectly long lens objective approaches it. It is recommended |
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to always first with lensThe para-focalparafocal, focusing with the safest |
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objective will then allow you to easily find your sample with another objective.objectives will facilitate locating the sample when switching to higher-magnification objectives.
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Lower the objective by pressing Home > Load Position
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Press Home>Load Position to lower the stage to its lowest positionPress Set Work Position to store this position If necessary,
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move slightly adjust the focus
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slightly up remove “Lower limit reached” Limit Reached message displayed on the
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touchscreentouch screen - Place the test slide on the microscope stage with the coverslip
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toward Always use the to perform the first focuswill significantly reduce the time required to set up the instrument. |
- If necessary,
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move so - to ensure that the sample is
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centered on Open the Zen software- centred under the objective
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On the computer:
the - the Locate tab, select BF or the desired fluorescence (Fluo, DAPI, GFP,
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DsRed DAPI, etc…- Cy5) to activate the configuration
- Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
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note| Info |
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Once calibrated, the focus |
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can be is typically found at Z = |
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1.5mm). The Z value can be found 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 |
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Home>Z-Positionunder 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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First Perform the initial focus |
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with lens before selecting another lens and continuing with secondary focusobjective before switching to higher-magnification objectives. |
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| title | Focusing with air objectives |
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| After performing the |
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first initial focus, on the microscope touch screen: - Press
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Home>Microscope>Turret>- Home>Microscope>Control>Objectives
- Press 10x, 20x
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or 40x - , 40x or 63x (0.75) to select the desired
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lens- objective
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| The 40x objective is the best Air objective because it has the |
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greatest number of optical corrections (Plan Apochromat) and the largest numerical aperture (0.95).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
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It offers a lateral resolution of 420nm at a wavelength of 550nm.- 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 off the illumination
Your sample is ready for acquisition! |
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lensesThis microscope does not have oil immersion . However, if there were, the procedure would be as follows: first initial focus, on the microscope touch screen:
- Press Home>Microscope>Turret>Objectives
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Press 63x Oil, 100x Oil (1.3) ou 100x Oil (1.4) to select the desired lens. The microscope will automatically lower the stage so that the sample is accessible.
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The 63x objective is the best oil objective because it has the most optical corrections (Plan Apochromat) and the largest numerical aperture .It offers a lateral resolution of 240nm at a wavelength of 550nmto select the desired objective. The microscope will automatically lower the stage so that the sample becomes accessible.
- Place a single drop of oil on
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your sample- the objective
- Press Done. The microscope will automatically return the
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sample - objective to its original position
In Zen |
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Blue software the - the Locate tab, select BF or the desired fluorescence (Fluo, DAPI, GFP, Cy3, DsRed,
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DAPI, etc…- Cy5) to activate the configuration
- Adjust the focus with the precision dial while looking through the eyepieces until the
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eyepieces until the - image is perfectly sharp
- In the Locate tab,
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select - select Off to turn off the illumination
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off
Your sample is ready for acquisition! |
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saved - 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).
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In any case, your files should be removed from the C: drive. |
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Close the Zen software- 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, clean oil objectives with lens cleaner and paper
- Select the lowest magnification objective and press load position to place the objectives in a safe position
If used, turn off the incubation module power
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strip Select the 10x objective and press load position to bring the objectives to the bottom positionstrip (#2A) and close the CO2 cylinder
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(#2B)(#2B) Turn off the microscope power bar (#3) - Turn off the computer
- Cover the instrument with the protective dust cover
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Important Reminders | - Take back your samples including ones in the microscope
- Leave the microscope and the working area clean
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idLightpath| title | Lightpath |
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The following schematics depict the light path for transmitted (bright-field and Phase Contrast) and reflected (fluorescence) lights. | PDF |
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| name | LightPath_Zeiss_Z1.pdf |
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Log| UI Expand |
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| - Measure Power output
- Modify camera power supply cable
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| title | 2025-10-22 Windows 11 and more |
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| - Installation of Windows 11
- Added CO2 connection to incubation module
- Cable management
- Replaced video card
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| title | 2025-09-01 Camera upgrade |
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| - Added compressed air connexion to the anti-vibration table
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| title | 2025-09-01 Camera upgrade |
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| - AxioCam replacement with PCO Edge 5.5
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Available manuals |
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| id | Log |
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title | LogTo do | - Check stage tilt
- Ask for Colibri Remote
- Ask for fluorescence backlight from LED
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| - Updated wiki with complete specifications
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| title | 2024-02-22 Microscope Firmware update |
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| - Microscope Firmware update to add Colibri to the touchscreen
- Zen 3.5 HotFix 10
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| - Computer replacement
- Added Colibri
- Added startup procedure
- Parafocality and paracentrality
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| - Added complete description
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idtitle inverted Light sources- Transmitted LED light
- Colibri 7 R(G/Y)B-UV 423052-9730-000 Serial 5440000661
Condenser |
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condenser Product number: TBD, Serial: TBDEmpty085 Air WD 8.8Stage
- Motorized stage Marzhauser Sensotech, Part number 432903-9011-000, #14 07 132052; 90-76-200-0820
- Remote control joystick
- Inserts
- Slide combo
- 6-well plate
- 35 mm dish
- Multi-well plat
Filters
- DAPI Filter Set 49
- GFP Filter Set 13
- Rhodamine Filter Set 43
- DHE (dihydroethidium) 424931
- Cy5 Filter Set 50
- Multiband FS90 HE LED
Detector
- Zeiss AxioCam MR R3 CCD Camera 1388 x 1040 pixels, 12-bit, 13 images/s at full resolution, detector size 8.9 mm x 6.7 mm. Model: r3.1 Part Number: 426509-9901-000. Serial: 1 22 12 5537
Workstation
Fujitsu Esprimo P920 E90+ WD 6.5Empty20x/0.5 Air Ph2 WD 2.040x/0.95 Air WD 0.25420941-9911-000
20x/0.5 Air Ph2 420351-9910-000
40x/0.75 Air Ph2 420361-9910-000
63x/0.75 Air Ph2 Long Distance 421381-9970-000
63x/1.4 Oil 420782-9900-000
Stage- Motorized stage Marzhauser Scan IM 130x100-2mm 90-24-550-0000 Serial 16053038
Stage Controller 90-76-024-1803 Serial 14 04 1 2040 SMC 2009 432929-9011-000 - Remote control joystick 2-Axis 90-76-200-0820 Zeiss Article 432903-9011-000 Serial 1615142059
- Inserts
- Slide combo
- Multi-well plate
Filters- DAPI Filter Set 49 488049-9901
- GFP Filter Set 13 488013-0000
- Rhodamine Filter Set 43 000000-1114-101
- DHE (dihydroethidium) 424931
- Cy5 Filter Set 50 488050-9901
- Multiband FS90 HE LED 489090-9110-000
Detector- Zeiss AxioCam MRm Model: r3.1 Part Number: 426509-9901-000. Serial: 1 22 12 5537
- PCO Edge 5.5 Model pco.API.Air.BX Serial 6000001404
Workstation- Computer
- Fujitsu Esprimo P920 E90+ Model MI5W Serial YLPS036541
- Motherboard
- Fujistsu D3222-A1 (1 PCIe Gen3 x16, 1 PCIe Gen 2 x16 (x4 lanes), 2x PCIe Gen2 x1
- Chipset Intel Q87
- BIOS UEFI compatible American Trend v4.6.5.4 R1.47.0 for D3222-A1x 2019-08-26
- Processor
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- Intel Core i5-4670 @ 3.4 GHz
- RAM
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32 GB DDR3 1600 MHz ECC (4 - 24 GB DDR3 800 MHz (3 x 8 GB)
- OS
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500 GB s Data Storage (2 x 1 TB spanned volume) Card AMD DDR5 dedicated memory- DDR5 0.768 TFLOPS 1x PCIe gen2 x16
- Monitor
- LG Flatron E2711 27' 1920 x 1080
- LG W2442 1920 x1080
- Software
- Zen Blue 3.5
- Serial 1121159628-524292
- HASP
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YRMDZ 1798977001 1798977001N- 1798977001
- Extended Focus
- Measurements
- Multi-Channel
- Panorama
- Software autofocus
- Time Series
- Z-stack
Incubation- Pecon stage top incubation
Anti-vibration table- TMC Model 63 512 Serial 933240
- Plain table top
Consumables |
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id | FAQ |
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| title | Troubleshooting & FAQ |
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| Troubleshooting| UI Expand |
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| title | I don't see any fluorescence! |
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| Technical support is free and unlimited. Contact us ! |
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| title | I see a high background in fluorescence |
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The fluoresncece light source is a Colibri while the transmitted light is a LED. What happens is that the fluorescence illumination reflects and into the LED and give a high background. To solve thisThis happens in the following conditions at - At 2.5x using the DAPI or GFP channel
- At 10x using the GFP channel
- At 20x using the GFP channel
To solve this you can use any of the following option: - Tilt the transmitted light arm backward
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or- Close the manual shutter between the condenser and the transmitted light
- Use a higher magnification objective
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Stop the light from entering the transmitted LED by using a cardboardFAQ| UI Expand |
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| title | Can I use this microscope to look at cell in a dish? |
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| Yes. This is an inverted microscope designed to look at specimen in a dish or a multi-well plate |
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. The objectives are optimized to image through thin glass bottom multi-well plates |
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. You may also image specimen mounted between a slide and a 0.17mm thick coverslip |
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| title | Can I use this microscope to perform timelapse experiments? |
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| Yes, but... This microscope has an incubation module to maintain temperature, humidity and gas. Yet it does not have a Definite focus which can maintain focus throughout time. Therefore, it is possible to loose the focus over long period |
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.. You can always do a software autofocus at different time but be aware of photo-toxicity if you are using fluorescence.
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| title | What is Extended Depth of Focus? |
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| Extended Focus is a method to flatten a Z-stack by keeping only the relevant information. In Zen: - Select the Processing Tab
- Select the Extended Depth of Focus Method
- Click Apply
- Wait until the processing is finished
- Save the processed image
Image AddedZ Stack animation Image AddedMaximum Intensity projection of a Z-Stack Image AddedExtended Depth of Focus projection of a Z-Stack
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| Tabs Container |
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| id | Demo Image |
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| title | Zeiss Axio-Observer Z1 |
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| direction | horizontal |
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| Tabs Page |
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| id | Demo Image |
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| title | Demo Image |
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Image Removed| Include Page |
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| Plateformes:_Plat-foot-en |
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| Plateformes:_Plat-foot-en |
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