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[🇫🇷 Version française|https://translate.google.com/translate?sl=en&tl=fr&u=https%3A%2F%2Fwiki.umontreal.ca%2Fspaces%2FMicroscopie%2Fpages%2F193498273%2FZeiss%2BZ1%2BColibri%2Ben]
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Zeiss

Axio

Z1-

Observer Z1

Colibri

inverted microscope

Microscope

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

Transmitted light

 

Applications

  • Inverted microscope
  • Widefield imaging
    • Brightfield
    • Phase contrast
    • Fluorescence
  • Incubation
  • Timelapse imaging
  • Deconvolution

Zeiss Z1 ColibriImage Removed

  • Extended Depth of Focus

Image Added

Image Added


Instrument Tabs
Tabs Container
Tabs Container
id
directionhorizontal
auitabspage
Tabs Page
idDescription
titleDescription

Description

Light sources

  • LED lamp for transmitted light

  • Colibri 7 R[G/Y]B-UV (385/469/555-590/631) for fluorescence
Expand
titleComplete specifications

Emission peak (nm)

Nominal Power (mW)Measured Power (mW)
385/30150 
469/38110 
555/3031 
631/3350 

View file
nameZeiss_Colibri-7

Objectives

_Product Information.pdf
height250

Image Added

Objectives

  1. 2.5x2.5x/0.075 Air

  2. 10x/0.25 Air Ph1
  3. 20x/0.5 Air Ph2
  4. 40x/0.75 Air Ph2

  5. 63x/0.75 Air Ph2 Long Distance
  6. 63x/1.4 Oil
Expand
titleComplete specifications
PositionNameBrandFull nameIDMagnificationNumerical ApertureImmersionTypeWorking distance (mm)Transmittance
(% [nm])
TechniqueCover glass thickness (mm)
12.5x/0.075 AirZeiss2.5x/0.075
EC Plan-Neofluar
M27
420320-9901-0002.5x0.075AirPlan Neofluar9.5>80% [400-840]BF, Fluo0.17
210x/0.25 AirZeiss10x/0.25 Ph1
N-Achroplan
M27 
420941-9911-00010x0.25AirN-AchroPlan

6.5

Not AvailableBF, PhC, Fluo0.17
320x/0.5 AirZeiss20x/0.50 Ph2
EC Plan-Neofluar
M27

420351-9910-000

20x0.50AirPlan Neofluar2.0Not AvailableBF, PhC, Fluo0.17
4

40x/0.75 Air

Zeiss40x/0.75 Ph2
EC Plan-Neofluar

M27

420361-9910-000

40x

0.75

Air

Plan Neofluar0.71Not AvailableBF, PhC, Fluo

0.17

563x/0.75 Air
Zeiss

63x/0.75 Corr Ph2
LD Plan-Neofluar

M27
421381-9970-000

63x

0.75

AirLD Plan-Neofluar1.7 at cover glass 0.75Not AvailableBF, PhC, Fluo

0 - 1.5

663x/1.4 OilZeiss63x/1.4 DIC
 Plan-Apochromat Oil
M27
420782-9900-00063x1.4OilPlan Apochromat0.19>80% [400-700]BF, Fluo0.17

Filter

  1. DAPI
  2. GFP
  3. Rhodamine
  4. DHE (dihydroethidium)
  5. Cy5
  6. Quadruple DAPI/GFP/Cy3/Cy5
Expand
titleComplete specifications
PositionNameBrandIDExcitation filterDichroic mirrorEmission filterComments
1DAPI
Filter Set 49
Zeiss

488049-9901

365/50
[325-375]
395LP445/50
[420-470]

2GFP
Filter Set 13
Zeiss488013-0000

470/20
[460-480]

495LP

517/25
[505-530]

3DsRed
Filter Set 43
Zeiss

000000-1114-101

545/25
[533-567]
570LP605/70
[570-640]

4DHECustom

Custom

500/50
[475-525]

540LP580/20
[570-590]
Undefined specifications
Best guess values
5Cy5
Filter Set 50
Zeiss488050-9901640/30
[625-655]
660LP

690/50
[665-715]


6Quadruple
DAPI/GFP/Cy3/Cy5
FS90 HE LED
Zeiss489090-9110-000


QBS
405+493+575+653

QBP 425/30+514/30+592/25+709/100

Excitation filters included in the light source FS90 HE LED

Detectors

  • PCO Edge 5.5Zeiss AxioCam MRm
    Expand
    titleComplete specifications
    Camera
Zeiss AxioCam MRm
  • PCO Edge 5.5

    Sensor Type

CCD
  • sCMOS

    Sensor Category

    Monochrome

    Nb Pixels

1
  • 5.
4
  • 5 M

    Pixel Layout

1388
  • 2560 x
1040
  • 2160

    Pixel size

     6.
45
  • 5 um

    Sensor size

8
  • 16.
9
  • 6 mm
x 6
  • x  14.
7
  • 0 mm

    Sensor diameter

11
  • 21.8 mm

    Bit depth

12
  • 16-bit
    Speed at full resolution
13
  • 100 images/s

    Max QE

 55
  •  60 % at 600 nm
Readout
  • Reading noise
 8
  •  1.0 e⁻

    Cooling

Pelletier
  • Forced air +7C

    Dark Current

    0.
7
  • 6 e⁻/pixel/sec

    Full well capacity

17
  • 30 000 e-

    Dynamic Range

    1:
2000
  • 30000

    Interface

FireWire (IEEE 1394a)
  • Dual Camera Link PCIe

    Mount

    C-mount
Image Removed
  • Image Added

    View file
    namePCO_Edge 5.5_Datasheet.pdf
    height250

  • Zeiss Axiocam MRm (not installed)
Expand
titleComplete specifications
sCMOS55 2560 21605 16 mm x  140 21.8 16100
CameraPCO Edge 5.5Zeiss Axiocam MRm

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 resolution13 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


Image RemovedImage Added

View file
nameZeiss_AxioCam MRm_Datasheet.pdf
height250

Tabs Page
idUser Guide
idUser Guide
titleUser Guide

User Guide

UI Expand
expandedtrue
titleStartup
  1. If not already done, turn on the the computer (#1) and use your UdeM credentials to log in to Windows using your UdeM credentials
  2. Remove the dust cover from the microscope
  3. If incubation is requiredturn on the incubation power bar (#2) on the desk near the computer and open the CO₂ cylinder (#2B) near the sink
    Note

    Make sure the humidifier is properly filled with distilled water.

  4. Turn on the microscope power bar bar (#3) on the desk near the computer

  5. 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.
  6. Start Zen

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).

  1. If Zen is open, close

  2. Zen
  3. it and wait

  4. for
  5. until it

  6. to close
  7. has completely shut down (this may take up to 30 seconds)

  8. On the Desktop, open the Softwares

  9.  folder
  10. folder

  11. Double-click Zen Settings for

  12. Axio-Osberver
  13. Zeiss Z1-Colibri

  14. A script will run and a black window will appear briefly

  15. Click OK to close the message
  16. When the message Settings for Zen have been imported successfully

  17. .
  18. appears, click OK to close it

  19. You can

  20. then
  21. now open

  22.  Zen
  23. Zen

UI Expand
titleFirst Use
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
UI Expand
titleLoading samples

During this procedure, you will:

  • Set
This procedure puts
  • the microscope
in
  • to a safe configuration
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.

title

On the microscope touch screen:

  • If not already done,
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
  • In Zen
:Once Zen software is
  • , once it has started a
popup
  • calibration dialog should
show up, just
  • appear. Simply click Calibrate Now.
    The microscope will lower the objectives, perform
a
  • an XY calibration
and then
  • first, followed by a Z calibration, and then return to its original position.
UI Expand
titleCalibration

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
Calibration within the Zen Software
Tip

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>Z

under 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
titleCalibration with the touch screen

side of the screen.

Expand
titleThe calibration dialog did not appear...

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 position
  • Press
    UI Expand
    titleVerifying if the system is already calibrated

    In Zen, within the Focus tab, located on the right side of the screen:

    1. Click Load to lower the objective

    2. The Z position is indicated below Current
    3. If Z position value is less than 100 um the system is calibrated

    On the microscope touch screen:

    1. Lower the objective by pressing Home > Load Position

    2. Press

    1. Set Work Position to store this position

    2. If necessary,

    move
    1. slightly adjust the focus

    slightly up
    1. upward to

    remove
    1. clear the Lower Z

    limit reached
    1. Limit Reached message displayed on the

    touchscreen
  • If not already done, press Home>Microscope>Control>Objectives>5x to select the 5x objective
  • If asked, tap Done to remove the oil lens cleaning warning
  • Press 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 be completed
  • Press 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 be completed
  • Note

    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

    UI Expand
    titleFirst focus
    1. touch screen

    2. The Z position value is indicated
    3. If the value is less than 0.1 mm then the system is calibrated
    UI Expand
    titleManual calibration with the software

    In Zen, within the Stage tab, located on the right side of the screen:

    1. If not already done, check the Show All option
    2. At the bottom of the tab click Calibrate

    3. A warning message will show up click Continue

    4. 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:

    1. If not already done, check the Show All option
    2. At the bottom of the tab click Calibrate

    3. A warning message will show up click Continue

    4. The microscope will then perform a Z calibration and then return to its original position

    XY and Z calibrations are now complete

    UI Expand
    titleManual calibration with the microscope touch screen
    Warning
    titleImportant
    Make sure to calibrate the focus before performing the first focus.

    On the microscope touch screen:

    1. If not already done, press Home>Microscope>Turret>Objectives>5x to select the 5x objective
      Info

      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.

    2. 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
    3. If necessary, move the focus slightly up to remove the Lower Z limit reached message displayed on the touchscreen
    4. Place the test slide on the microscope stage with the coverslip toward the objective
      Note
      titleImportant

      Always use the test slide to perform the first focus.

    5. 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 configuration
  • Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
    Note

    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 illumination
    1. Navigate to Home > Microscope > XYZ > Position > Z-Position > Set Zero > Auto to perform focus calibration

    2. Press OK to start the calibration procedure

    3. Wait a few seconds for the calibration to complete

    4. Lower the objective by pressing Home > Load Position

    5. Press Set Work Position to store this position

    6. If necessary, slightly adjust the focus upward to clear the Lower Z Limit Reached message displayed on the touch screen

    7. Navigate to Home > Microscope > XYZ > Position > XY-Position > Set Zero > Auto to perform a stage calibration

    8. Press OK to start the calibration procedure

    9. Wait a few seconds for the calibration to complete

    XY and Z calibrations are now complete

    UI Expand
    titleEnsuring the calibration dialog is displayed at startup

    In Zen:

    1. In the menu bar, navigate to Tools > Options

    2. Select Startup/Shutdown

    3. Under Stage/Focus Calibration, ensure Request Stage/Focus Calibration on Startup is checked

    4. Click OK to close the Options dialog

    UI Expand
    titleInitial focus
    Warning
    titleImportant
    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:

    1. If not already done, select the lowest-magnification objective Home > Microscope > Control > Objectives > 2.5x
    Info

    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.

    1. Lower the objective by pressing Home > Load Position

    2. Press Set Work Position to store this position

    3. If necessary, slightly adjust the focus upward to clear the Lower Z Limit Reached message displayed on the touch screen

    4. Place the test slide on the microscope stage with the coverslip facing the objective
      Note
      titleImportant

      Using a test slide will significantly reduce the time required to set up the instrument.

    5. If necessary, adjust the stage to ensure that the sample is centred under the objective

    In Zen:

    1. In the Locate tab, select BF or the desired fluorescence (Fluo, DAPI, GFP, Cy3, Cy5) to activate the configuration
    2. Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
    Info

    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.

    1. In the Locate tab, select Off to turn off the illumination
    UI Expand
    titleSeconday focus
    Warning
    titleImportant

    Perform the initial focus using the safest objective before switching to higher-magnification objectives.

    UI Expand
    titleFocusing with air objectives

    After performing the initial

    UI Expand
    titleSeconday focus
    Warning
    titleImportant

    First focus with the safest objective before selecting another lens and continuing with secondary focus.

    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
    UI Expand
    titleFocusing with air objectives
    Info
    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

    There are two (2) 40x objectives, make sure you select the Air 40x

    In Zen :

    1. In the Locate tab, select BF or the desired fluorescence (DAPI, GFP, mPlum) to activate the configuration
    2. Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
    3. In the Locate tab, select Off to turn the illumination off
    4. Your sample is ready for acquisition!
    UI Expand
    titleFocusing with oil objectives

    After performing the first focus, on the microscope touch screen:

    1. Press
    2. Home>Microscope>Turret>
    3. Home>Microscope>Control>Objectives
    4. Press
    5. 63x Oil
    6. 10x, 20x,
    7. 40x Oil
    8.  40x or 63x (0.75) to select the desired objective
    9. . The microscope will automatically lower the stage so that the sample is accessible.
    Info
    The 40x
    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).

    In Zen:

    1. In the Locate tab, select BF or the desired fluorescence (Fluo, DAPI, GFP, Cy3, DsRed, Cy5) to activate the configuration
    2. Adjust the
    Note

    There are two (2) 40x objectives, make sure you select the 40x Oil

  • Place a single drop of oil on the objective
  • Press Done. The microscope will automatically return the objective to its original position
  • In Zen :

    1. In the Locate tab, select BF or the desired fluorescence (DAPI, GFP, mPlum) to activate the configuration
    2. Adjust the
    3. focus with the precision dial while looking through the eyepieces
    4. until the
    5. until the image is perfectly sharp
    6. In the Locate tab,
    7. select
    8. select Off to turn off the illumination
    9. off

    Your sample is ready for acquisition!

    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
    titleFocus Calibration Z
    On the microscope 
    UI Expand
    title
    Loading samples
    Focusing with oil objectives

    After performing the initial focus, on the microscope

    touch screen:

    1. 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
  • Press Home>Microscope>Control>Objectives>2.5x to select the 2.5x objective
  • If asked, tap Done to remove the oil lens cleaning warning
  • Press Home>Microscope>XYZ>Position>Z-Position>Set zero>Auto to perform focus calibration
  • Press OK to start the focus calibration procedure
  • Wait a few seconds for the calibration to be completed
  • Note

    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
    titleFirst focus
    Warning
    titleImportant

    Make sure to calibrate the focus before performing the first focus.

    On the microscope touch screen:

    1. Press Home>Microscope>Turret>Objectives
    2. Press 2.5x to select the 2.5x lens
      Info

      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.

    3. Press Home>Load Position to lower the objectives to its lowest position
    4. Press Set Work Position to store this position
    5. If necessary, move the focus slightly up to remove the Lower Z limit reached message displayed on the touchscreen
    6. Place the test slide on the microscope stage with the coverslip toward the objective
      Note
      titleImportant

      Always use the test slide to perform the first focus.

    7. 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, Rhodamine, etc…) to activate the configuration
  • Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
    Note

    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
    titleSecondary focus
    Warning
    titleImportant

    First focus with the safest lens before selecting another lens and continuing with secondary focus.

    UI Expand
    titleFocusing with air objectives

    After performing the first focus, on the microscope touch screen:

  • Press Home>Microscope>Control>Objectives
  • Press 20x or 40x or 63x to select the desired lens
    Info

    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 sharp
  • Your sample is ready for acquisition!
  • UI Expand
    titleFocusing with oil lenses

    After performing the first focus, on the microscope touch screen:

    1. Press Home>Microscope>Control>Objectives
    2. Press 63x Oil. The microscope will automatically lower the objective so that the sample is accessible

    3. Place a drop of oil on your sample
    4. Press Done. The microscope will automatically return the sample to its original position

    In Zen:

    1. In the Locate tab, select BF or the desired fluorescence (DAPI, GFP, Rhodamine, etc…) to activate the configuration
    2. Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
    3. In the Locate tab, select Off to turn the illumination off
    4. Your sample is ready for acquisition!
    UI Expand
    titleStorage management
    • 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

    In any case, your files should be removed from the C: drive.

    UI Expand
    titleShutdown
    1. Save your data
    2. Close Zen
    3. Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
    4. If used, turn off the incubation module power strip (#2A) and close the CO2 cylinder (#2B)

    5. Select the 2.5x objective and press load position to bring the objectives to the bottom position

    6. Turn off the microscope power bar (#3)

    7. Turn off the computer
    8. Cover the instrument with the protective dust cover
    Note
    titleImportant Reminders
    • Take back your samples including ones in the microscope
    • Leave the microscope and the working area clean
    Tabs Page
    idLightpath
    titleLightpath

    The following schematics depict the light path for transmitted (bright-field and Phase Contrast) and reflected (fluorescence) lights.

    View file
    nameZeiss_Z1-Colibri_LightPath.pdf
    height250

    Tabs Page
    idManuals
    titleManuals

    Available manuals

    Tabs Page
    idLog
    titleLog
    UI Expand
    title2024-02-22 Microscope Firmware update
    • Microscope Firmware update to add Colibri to the touchscreen
    • Zen 3.5 HotFix 10
    UI Expand
    title2022-05-09
    • Computer replacement
    • Added Colibri
    • Added startup procedure
    • Parafocality and paracentrality
    UI Expand
    title2022-03-17
    • Added complete description
    UI Expand
    title2021-09-27
    • Added to wiki
    1. Home>Microscope>Turret>Objectives
    2. Press 63x Oil (1.4)to select the desired objective. The microscope will automatically lower the stage so that the sample becomes accessible.

    3. Place a single drop of oil on the objective
    4. Press Done. The microscope will automatically return the objective to its original position

    In Zen :

    1. In the Locate tab, select BF or the desired fluorescence (Fluo, DAPI, GFP, Cy3, DsRed, Cy5) to activate the configuration
    2. Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
    3. In the Locate tab, select Off to turn off the illumination

    Your sample is ready for acquisition!

    UI Expand
    titleStorage management
    • 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

    In any case, your files should be removed from the C: drive.

    UI Expand
    titleShutdown
    1. Save your data
    2. Close Zen
    3. Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
    4. If used, clean oil objectives with lens cleaner and paper
    5. Select the lowest magnification objective and press load position to place the objectives in a safe position
    6. If used, turn off the incubation module power strip (#2A) and close the CO2 cylinder (#2B)

    7. Turn off the microscope power bar (#3)

    8. Turn off the computer
    9. Cover the instrument with the protective dust cover
    Note
    titleReminder
    • Take back your samples including ones in the microscope
    • Leave the microscope and the working area clean

    Tabs Page
    titleLog

    Log

    UI Expand
    titleTo be done
    • Measure Power output
    • Modify camera power supply cable
    UI Expand
    title2025-10-22 Windows 11 and more
    • Installation of Windows 11
    • Added CO2 connection to incubation module
    • Cable management
    • Replaced video card
    UI Expand
    title2025-09-01 Camera upgrade
    • Added compressed air connexion to the anti-vibration table
    UI Expand
    title2025-09-01 Camera upgrade
    • AxioCam replacement with PCO Edge 5.5
    UI Expand
    title2025-08-21 Wiki Updated
    • Updated wiki with complete specifications
    UI Expand
    title2024-02-22 Microscope Firmware update
    • Microscope Firmware update to add Colibri to the touchscreen
    • Zen 3.5 HotFix 10
    UI Expand
    title2022-05-09
    • Computer replacement
    • Added Colibri
    • Added startup procedure
    • Parafocality and paracentrality
    UI Expand
    title2022-03-17
    • Added complete description
    UI Expand
    title2021-09-27
    • Added to wiki
    Tabs Page
    titleTechnical Datasheet

    Technical Datasheet

    Stand

    • Zeiss Axio-Observer Z1 inverted Serial: 3851001242 Part Number: 431007-9902-000
      System ID: 1024979772
    • Camera adapter Model 60N-C, 1", 1x, Model: 426114

    Light sources

    • Transmitted LED light
    • Colibri 7 R(G/Y)B-UV 423052-9730-000 Serial 5440000661

    Condenser

    • Manual condenser
    • Lens NA 0.35 WD 70 mm Part Number: 424241

    • Filter turret 6 positions manual

      1. H

      2. Ph0
      3. Ph1

      4. Ph2

      5. DIC

      6. DUC

    Objectives

    1. 2.5x/0.075 Air 420320-9901-000

    2. 10x/0.25 Air Ph1 420941-9911-000

    3. 20x/0.5 Air Ph2 420351-9910-000

    4. 40x/0.75 Air Ph2 420361-9910-000

    5. 63x/0.75 Air Ph2 Long Distance 421381-9970-000

    6. 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

    1. DAPI Filter Set 49 488049-9901
    2. GFP Filter Set 13 488013-0000
    3. Rhodamine Filter Set 43 000000-1114-101
    4. DHE (dihydroethidium) 424931
    5. Cy5 Filter Set 50 488050-9901
    6. 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
      • Intel Core i5-4670 @ 3.4 GHz
    • RAM
      • 24 GB DDR3 800 MHz (3 x 8 GB)
    • OS Drive
      • 1 TB SSD 550 MB/s
      • Win 11 23H2
    • Storage
      • 2 TB HD 110 MB/s
    • Video Card
      • AMD FirePro V4900 1 GB 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 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

    • CO2 Tank
    • Oil
    • Lens Cleaner

    Manuals

    Tabs Page
    titleTroubleshooting & FAQ

    Troubleshooting

    UI Expand
    titleI don't see any fluorescence!

    Technical support is free and unlimited. Contact us !

    UI Expand
    titleI see a high background in fluorescence

    This 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
    • Close the manual shutter between the condenser and the transmitted light
    • Use a higher magnification objective

    FAQ

    UI Expand
    titleCan I use this microscope to look at cell in a dish?

    Yes. This is an inverted microscope designed to look at specimen in a dish or a multi-well plate. The objectives are optimized to image through thin glass bottom multi-well plates. You may also image specimen mounted between a slide and a 0.17mm thick coverslip. For long timelapse, be aware of photo-toxicity.

    Tabs Page
    idTechnical Datasheet
    titleTechnical Datasheet

    Stand

    • Zeiss Axio-Observer Z1 inverted  Serial: 3851001242 Part Number: 431007-9902-000
      System ID: 1024979772
    • Camera adapter Model 60N-C, 1", 1x, Model: 426114

    Light sources

    • Transmitted LED light
    • Colibri 7 R(G/Y)B-UV 423052-9730-000 Serial 5440000661

    Condenser

    • Manual condenser Product number: TBD, Serial: TBD
    • Lens NA 0.35 WD 70 mm Part Number: 424241

    • Filter turret 6 positions manual

      1. H

      2. Ph0
      3. Ph1

      4. Ph2

      5. DIC

      6. DUC

    Objectives

    1. 2.5x/0.075 Air 420320-9901-000

    2. 10x/0.25 Air Ph1 420941-9911-000

    3. 20x/0.5 Air Ph2 420351-9910-000

    4. 40x/0.75 Air Ph2 420361-9910-000

    5. 63x/0.75 Air Ph2 Long Distance 421381-9970-000

    6. 63x/1.4 Oil 420782-9900-000

    Stage

    • 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

    1. DAPI Filter Set 49 488049-9901
    2. GFP Filter Set 13 488013-0000
    3. Rhodamine Filter Set 43 000000-1114-101
    4. DHE (dihydroethidium) 424931
    5. Cy5 Filter Set 50 488050-9901
    6. Multiband FS90 HE LED 489090-9110-000

    Detector

    • Zeiss AxioCam MR R3 Model: r3.1 Part Number: 426509-9901-000. Serial: 1 22 12 5537

    Workstation

    • Fujitsu Esprimo P920 E90+
    • Intel Core i5-4670 @ 3.4 GHz
    • RAM 32 GB DDR3 1600 MHz ECC (4 x 8 GB)
    • OS 500 GB SSD 550 MB/s
    • 2 TB HD Data Storage (2 x 1 TB spanned volume) 110 MB/s
    • Video Card AMD FirePro V4900 1 GB DDR5 dedicated memory
    • Monitor LG Flatron E2711 27' 1920 x 1080
    • Software Zen Blue 3.5 SN=1121159628-524292 HASP=1798977001

    Incubation

    • Pecon stage top incubation

    Consumables

    • CO2 Tank
    • N2 Tank
    • Oil
    • Lens Cleaner
    Tabs Page
    idFAQ
    titleTroubleshooting & FAQ

    Troubleshooting

    UI Expand
    titleI see a high background in fluorescence

    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 this:

    • Tilt the transmitted light arm backward
      or
    • Stop the light from entering the transmitted LED by using a cardboard
    FAQ

    UI Expand
    titleCan I use this microscope to look at cell in a dishperform timelapse experiments?

    Yes, but... This

    is an inverted microscope designed to look at specimen in a dish or a multi-well plate
  • The objectives are optimized to image through thin glass bottom multi-well plates
  • You may also image specimen mounted between a slide and a 0.17mm thick coverslip
  • For long timelapse,

    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. You can always do a software autofocus at different time but be aware of photo-toxicity if you are using fluorescence.

    UI Expand
    titleCan I use this microscope to perform timelapse experiments?What is Extended Depth of Focus?

    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

    Z Stack animationImage AddedMaximum Intensity projection of a Z-StackImage AddedExtended Depth of Focus projection of a Z-StackImage Added

    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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