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Button Hyperlink
iconbuild
titleFrançais
typeprimary
urlhttps://wiki.umontreal.ca/pages/viewpage.action?pageId=193498625

Tabs Container
idInstrument Tabs
titleZeiss Axio-Observer Z1 Spinning diskImager Z2
directionhorizontal
Tabs Page
idDescription
titleDescription

Zeiss Axio-Observer

Z1

spinning-disk

confocal

microscope

J-A Bombardier Building, Room 3223-101
Advanced Microscope Tier 2 usage price

Instrument awarded to Dr. Daniel Zenklusen by the Canadian Foundation for Innovation (CFI)

  • Applications
    • Transmitted light
    • Interference Contrast (DIC)
    • Fluorescence
  • Light sources

    • 12V 100W halogen lamp for transmitted light

    • Lumencor SOLA for visible fluorescence

      Développer
      titleLumencor SOLA complete specifications

      Emission peak (nm)

      Power (mW)

      334

      5

      365

      20

      405

      19

      436

      22

      546

      21

      579

      18

  • Objectives

  1. 10x/0.30 Air WD 5.30

  2. 63x/1.40 Oil WD 0.19

  3. 100x/1.40 Oil WD 0.17

  4. TIRF adjustment

  5. TIRF adjusment

  6. Empty


Développer
titleFull lens specifications

Position

Nom

Marque

Nom complet

Identifiant

Grossissement

Ouverture numérique

Immersion

Type

Distance de travail (mm)

Transmittance

(% [nm])

Technique

Épaisseur du couvre-objet (mm)

1

10x/0.30
Air

Zeiss

10x/0.3 DIC I

EC Plan-Neo Fluar

M27

420340-9901-000

10x

0.3

Air

Plan Neofluar

5.2

>90% [480-780]

BF, DIC, Fluo

0.17

2

Empty

Zeiss

20x/0.8 DIC II

Plan-Apochromat
W0.8x1/36"

440640-9903-000

20x

0.8

Air

Plan Apochromat

0.55

>90% [410-800]

BF, DIC, Fluo

0.17

3

Empty


Zeiss

40x/0.75 DIC II
EC Plan-Neofluar
M27

420360-9900-000

40x

0.75


Air


Plan Neofluar

0.71

>90% [410-780]

BF, DIC, Fluo

0.17

4

63x/1.40

Huile

Zeiss

63x/1.4 DIC III

Plan-Apochromat

M27

420782-9900-000

63x

1.4



Remarque
Huile



Plan Apochromat

0.19

>80% [440-710]

BF, DIC, Fluo

0.17

5

Empty












6

100x/1.40
Huile

Zeiss

100x/1.4 DIC III
Plan-Apochromat
M27

420792-9900-000

100x

1.4



Remarque
Huile



Plan Apochromat

0.17

>80% [400-820]

BF, DIC, Fluo

0.17

BF: Bright-field
DIC: Interference contrast

  • Filter cubes
  1. DAPI
  2. BP405
  3. 76
  4. 77
  5. Empty
    Développer
    titleComplete filter specifications


    Position

    Nom

    Marque

    Identifiant

    Filtre d'excitation 

    Miroir dichroïque

    Filtre d'émission

    Commentaire



    1

    DAPI

    Filter Set 49

    Zeiss

    488049-9901-000

    365/50

    [340-390]

    395LP

    445/50

    [420-470]




    2

    GFP

    Filter Set 38

    Zeiss

    000000-1031-346

    470/40
    [450-490]

    495LP

    525/50

    [500-550]


    FT 495

    BP 525/50

    3

    YFP

    Filter Set 46

    Zeiss

    000000-1196-681

    500/20

    [490-510]

    515LP

    535/30

    [520-550]




    4

    DsRed

    Filter Set 43

    Zeiss

    000000-1114-101

    545/25

    [533-557]

    570LP

    605/70

    [570-640]

    FT 570

    BP 605/70


    5

    TxRed

    Filter Set 45

    Zeiss

    000000-1114-462

    560/40

    [540-580]

    585LP

    630/75

    [593-667]




    6

    Cy3.0

    Chroma

    SP102v1

    546/11
    [541-551]

    557LP

    567/15

    [560-574]




    7

    Cy3.5

    Chroma

    SP103v1

    581/10
    [576-586]

    593LP

    617/40

    [597-637]




    8

    Cy5 narrow

    Chroma

    49009

    640/30
    [625-655]

    660LP

    690/50

    [665-715]




    9

    Analyseur DIC

    Zeiss








    10

    Vide









  • Detector
    • 2 x Evolve512  camera from Photometrics


Tabs Page
idUser Guide
titleUser Guide


UI Expand
expandedtrue
titleStartup
  1. Turn on the computer (#1)
  2. If required, turn on the incubation power bar
  3. Turn on the camera and laser power bar on the left of the microscope (#2)

  4. Turn on the microscope power bar on the right of the microscope (#3)

  5. Use your UdM credentials to log in to Windows

    Remarque

    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.

  6. Start the Zen Blue software


UI Expand
titleFirst Use

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.

Remarque

Please note, this procedure will delete all your experiment protocols and restore the software to its original settings.

  1. If open, close the Zen Blue software and wait for it to close completely (up to 30 seconds)
  2. On the Desktop open the Documentation folder
  3. Double-click Settings for Axio-Observer Z1
  4. A script will run and a black window will appear briefly
  5. You can then reopen the Zen Blue software


UI Expand
titleLoading samples

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 touch screen:

  1. Press Home>Load Position to lower the stage to its lowest position
  2. 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. Press Home>Microscope>Turret>Objectives>10x to select the 10x objective
  5. If asked, tap Done to remove the oil lens cleaning warning
  6. Press Home>Microscope>XYZ>Position>Z-Position>Set zero>Auto to perform focus calibration
  7. Press OK to start the focus calibration procedure
  8. Wait a few seconds for the calibration to be completed
Remarque

Once calibrated, the focus can be found at Z = 5 mm). The Z value can be found on the microscope touch screen Home>Z-Position


UI Expand
titleFirst focus
Avertissement
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 10x to select the 10x lens
    Info

    The 10x objective is the safest because it has the longest working distance (5.3 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 stage 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
    Remarque
    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:

  1. Open the Zen Blue software
  2. In the Locate tab, select BF or the desired fluorescence (GFP, DsRed, DAPI, etc…) to activate the configuration
  3. Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp

    Remarque

    Once calibrated, the focus can be found at Z = 5 mm). The Z value can be found on the microscope touch screen Home>Z-Position


  4. In the Locate tab, select Off to turn off the illumination


UI Expand
titleSeconday focus
Avertissement
titleImportant

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


UI Expand
titleFocusing with air objectives

This microscope does not have additional air objectives. However, if there were, the procedure would be as follows:

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

  1. Press Home>Microscope>Turret>Objectives
  2. Press on the desired lens
  3. Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
  4. 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>Turret>Objectives
  2. Press 63x Oil (1.4)or 100x Oil (1.4) to select the desired lens. The microscope will automatically lower the stage so that the sample is accessible.

    Info

    The 63x objective is the best oil objective because it has the most optical corrections (Plan Apochromat) and the largest numerical aperture (1.4). It offers a lateral resolution of 240nm at a wavelength of 550nm.


  3. Remove your sample from the microscope
  4. Place a drop of oil on the objective
  5. Replace your sample from the microscope
  6. Press Done. The microscope will automatically return the sample to its original position

In Zen Blue software:

  1. In the Locate tab, select BF or the desired fluorescence (GFP, DsRed, DAPI, 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).
Remarque

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

UI Expand
titleShutdown
  1. Save your data
  2. Close the software Zen Blue
  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 lenses with lens cleaner and paper
  5. Turn off the microscope power bar on the right of the microscope (#3)

  6. Turn off the camera and laser power bar on the left of the microscope (#2)
  7. Turn off the computer
Remarque
titleImportant Reminders
  • Take back your samples including ones in the microscope
  • Leave the microscope and the working area clean
Tabs Page
idLight Path
titleLight Path

The following diagrams allow you to follow the light path in transmitted light (bright field, DIC) and in reflected light (fluorescence).





_blank
Tabs Page
idManuals
titleManuals
  • Zen quick start guide.pdf
  • See

    Link in New Window
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    hrefhttps://biochimie.umontreal.ca/plateformes-scientifiques-bmm/microscopie/microscope-zeiss-axio-observer-z1-avec-disque-rotatif/
    target_blank
    on the
    Link in New Window
    iconfalse
    linkTextBiochemistry Department Microscopy Platform
    hrefhttps://biochimie.umontreal.ca/plateformes-scientifiques-bmm/microscopie/
    target_blank

  • Training and booking available upon request to the corresponding platform manager

  • Consult the
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    hrefhttps://biochimie.umontreal.ca/wp-content/uploads/sites/37/2021/02/Tarifs_2021-22_pour_utilisation_des_plateformes_BMM.pdf
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    and
    Link in New Window
    iconfalse
    linkTextaccess policies
    hrefhttps://biochimie.umontreal.ca/wp-content/uploads/sites/37/2021/10/Reglements_microscopes_oct_2021.pdf
    target_blank
    of the
Link in New Window
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linkTextBiochemistry Department Microscopy Platform
hrefhttps://biochimie.umontreal.ca/plateformes-scientifiques-bmm/microscopie/
target



Tabs Page
idLog
titleLog


UI Expand
titleTo do
Check Loss of 488nm laser power


UI Expand
title2021-11-20
  • Zen 2.6 updated hotfix 12
  • Microsoft Windows update
  • Laser output measured in spinning disk mode at the sample with 10x objective, 1x Extended tubelens, 100% laser power and ALL lasers ON:
    • 405nm: 1.08 mW StdDev 0.01mW
    • 488nm: 3.3 mW  StdDev <0.01mW (3.7 mW if laser 405 is OFF)

      Avertissement
      title405nm and 488nm Interactions

      When both 405nm and 488nm laser are ON simultaneously, the output power is decreased by 11%.


    • 561nm: 2.6 mW StdDev <0.01mW
    • 639nm: 1.37 mW StDev 0.02 mW
  • Control for vibrations during construction work on the 4th floor


UI Expand
title2021-11-03
  • Error when the definite focus is used (unexpected error the definite focus did not respond)
  • To solve it:
    • Turn off the Zen Software
    • Turn off the microscope power bar (#3)
    • Turn off the definite focus by pressing and holding the definite focus button
    • Turn on the definite focus by pressing once the definite focus button
    • Wait until the definite focus display shows "Detecting stand"
    • Turn on the microscope power bar (#3)
  • Confirm that the definite focus is showing in the microscope tactile display
  • Open Zen software and run an experiment using the definite focus to confirm it is fully functional


UI Expand
title2021-09-27
  • Added to wiki
Tabs Page
idTechnical Datasheet
titleTechnical Datasheet

Stand

  • Zeiss AxioObserver Z1 upright Serial:
    Part Number: 
    System ID
  • Camera adapter Model 60N-C, 1", 1x, Model: 426114

  • Motorized Neutral density filters for transmitted light

  • Manual Field diaphragm for transmitted light

  • Manual polarizer
  • Left imaging port with manual splitter camera adapter Model 60N-C, 1", 1x, Model: 426114
  • Trinocular with 100% ocular 40% occular/70% camera and 100% manual splitter
  • 3mm liquid light guide #805-0038
  • Zeiss 423302-0000 Collimator
  • Motorized Aperture diaphragm
  • Motorized Fluorescence field diaphragm

Light sources

  • Transmitted Halogen light 12V 100W HAL 100 #423000
    • TBD Filters
  • Lumencor SOLA Serial 

Condenser

  • Motorized condenser #424201-9902
  • Lens NA 0.9 WD TBD Part Number: TBD

  • Manual polarizer
  • Filter turret 6 positions manual

    1. H Empty

    2. D Darkfield
    3. DIC III #426706
    4. Ph3
    5. DICII #426702
    6. Ph 2

    7. DIC 426701
    8. Ph 1

Objectives

  1. 10x/0.30 Air WD 5.30 DIC I  Plan-NeoFluar M27 420340-9901-000

  2. 63x/1.40 Oil WD 0.19 DIC III Plan-Apochromat M27 420782-9900-000

  3. 100x/1.40 Oil WD 0.17 DIC III Plan-Apochromat M27 420792-9900-000 

Stage

  • Motorized stage Zeiss AIM System #2502000124
  • Remote control joystick
  • Inserts
    • Slide only
    • Plate

Filters

10-positions motorized filter wheel #

  1. DAPI Zeiss Filter Set 49 cube 424933

Detector

  • 2 camera Evolve 512 Serial

Workstation

  • HP Z800 Workstation Serial: CZC1473Y0Q Part number: WJ112ECJ#AK6
  • 2 x Intel Xeon X5650 2.66 GHz
  • RAM 24 GB DDR3 1333 MHz ECC (12 x 2 GB)
  • OS 500 GB SSD 410 MB/s
  • 2 TB HD Data Storage (2 x 1 TB spanned volume) 110 MB/s
  • Video Card ATI FirePro V5800 1 GB DDR5
  • Monitor Dell ST2410  24' 1920 x 1080
  • Software Zen Blue 2.6 Hotfix 12

Incubation

  • Zeiss Incubation

Consumables


Tabs Page
idFAQ
titleTroubleshooting & FAQ

Troubleshooting

UI Expand
Troubleshooting
titleI don't see any fluorescence!

The best way to solve a problem in Microscopy is to follow the light path. You will find in the Light path tab of this page, the diagrams which will allow you to follow the light all along its path through the microscope.

  1. Open the light path file
  2. Starting from the light source and moving towards the detector, verify that there is indeed light after each component of the microscope

FAQ

UI Expand
titleFAQCan 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.



Tabs Container
idDemo Image
titleZeiss Axio-Observer Z1 Spinning disk
directionhorizontal


Tabs Page
idDemo Image
titleDemo Image





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