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Button Hyperlink
iconbuild
titleFrançais
typeprimary
urlhttps://wiki.umontreal.ca/display/Microscopie/Nikon+Ti2-E

Tabs Container
idInstrument Tabs
titleNikon Ti2-E
directionhorizontal
Tabs Page
idDescription
titleDescription

Nikon Ti2-Fura

inverted

microscope

Desmarais Building, Room 2236
Instrument awarded to the CI2B by the Canadian Foundation for Innovation (CFI)
Advanced Microscope Tier 1 usage price

 

Applications

Transmitted light, Bright-field

  • Widefield
    • Brightfield

    • Fluorescence

  • Live imaging

  • Time-lapse imaging
    • Calcium ratiometric imaging



    Tabs Container
    directionhorizontal
    Tabs Page
    idDescription
    titleDescription

    Light sources

    • LED lamp for transmitted light

    • CoolLed pE340-fura

    • Lumencor Spectra

    Objectives

    1. 4x/0.2 Air
    WD 20
    2. 10x/0.45 Air
    3. 20x/0.75 Air
    DIC WD 1.0
    2. 40x
    3. 60x/1.4 Oil DIC WD 0.13
    4. Empty
    Expand
    title
    Objectives complete
    Complete specifications
    PositionNameBrandFull nameIdentifierWorking distance (mm)Transmittance
    (% [nm])    		TechniquesCover glass thickness (mm)
    14x/0.2
    Air    		Nikon

    4x/0.2 Air
    CFI Plan

    Apo

    Apochromat Lambda

    MRD00045

    		20.0>80% [400-1000]BF, Fluo0.17
    210x/0.45
    Air    		Nikon

    10x/0.45 Air
    CFI Plan Apochromat Lambda

    		MRD00105

    4.0

    		 DIC N1
    320x/0.75
    Air
    DIC
    Nikon20x/0.75 Air
    CFI Plan
    Apo
    Apochromat Lambda
    DIC N2
    MRD00205

    1.0

    		>80% [400-950]BF, Pol, DIC, Fluo
    DIC N2    		0.17
    440x/0.75
    Air    		Nikon40x/0.75 Air
    Plan Fluor     		MRH00400

    0.72

    		 

    5

    60x/1.4
    Oil

    DIC

    		Nikon60x/1.4 Oil
    CFI Plan
    Apo
    Apochromat Lambda
    DIC N2
    MRD016050.13>80% [475-725]BF, Pol, DIC, Fluo
    DIC N2
    6

    Empty





    		 

    Filters

    1. DAPI

    2. GFP

    3. Cy3

    4. Cy5
    5. Fura
    Expand
    title
    Filters complete
    Complete specifications
    PositionNameBrandIDExcitation FilterDichroic mirrorEmission FilterComments
    1DAPINikonDAPI-U HQ395/25x
    [383-408]    		425LP460/50m
    [435-485]    		C-FL-C DAPI-U HQ
    2GFPSemrockGFP-4050B-000

    466/40x
    [446-486]

    495LP

    		525/50m

    [500-550]    		Nikon ID 96372
    3Cy3






    4Cy5Semrock

    Cy5-5070A

    		617/55x
    [590-645]    		652LP697/77m
    [659-736]    		Nikon ID 96376
    5Fura






    6Empty






    Detector

    • Photometrics Prime 95B

    25mm CMOS Monochrome Camera  x  pixels, 16-bit, 30 images/s at full frame (100 images/s at full frame with camera link connector)

    • 25mm 

    Expand
    titleComplete specifications

    Camera

    		Photometrics Prime 95B

    Sensor Type

    		Back-illuminated sCMOS

    Sensor Category

    		Monochrome

    Nb Pixels

    		2.6 M

    Pixel Layout

    		1608 x 1608

    Pixel size

    		11.0 um

    Sensor size

    		17.7 mm x 17.7 mm 

    Sensor diagonal

    		25 mm

    Bit depth

    		16-bit
    Speed at full resolution30 images/s

    Max QE

    		95 %
    Readout noise1.6 e⁻

    Cooling

    		-20°C Air

    Dark Current

    		0.55 e⁻/pixel/sec

    Full well capacity

    		80 000 e-

    Dynamic Range

    		1: 50 000

    Interface

    		USB 3.0

    Mount

    		F-mount

    Image Added

    View file
    namePhotometrics_Prime-95B_Datasheet.pdf
    height250
    View file
    namePhotometrics_Prime-95B_Manual.pdf
    height250

    Tabs Page
    idUser Guide
    titleUser Guide
    UI Expand
    expandedtrue
    title
    Startup
    Start-up
    1. If not already done, turn
    Turn Start the software
    1. on the computer (#1) and log in to Windows using your UdeM credentials
    2. Remove the dust cover from the microscope
    3. Turn on the microscope power bar (#2) on the left of the computer
    Use your UdeM credentials to log in to Windows
    Info

    The first time you use the instrument, you need to import the microscope settings into the software. To do this follow the instructions First use protocol.

    1. 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.

    2. Start-up NIS-Elements
    UI Expand
    expandedtrue
    titleFirst Use

    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.

    Note

    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).

    1. If open, close NIS-Elements and wait until it is completely closed (up to 30 seconds)
    2. On your Desktop open the Softwares folder
    3. Open NIS Settings Utility
    4. Click on the Import tab
    5. Click on Browse
    6. Navigate to your C:\Users\Public\Documents
    7. Select the file Nikon_Ti2-Fura_NIS Settings.bin
    8. Click Select
    9. Select all items
    10. Click Import
    11. Click OK
    12. Close the NIS Settings Utility
    13. You can now reopen 
    1. NIS-Elements
    UI Expand
    titleLoading samples

    During this procedure, you will:

    • Set the microscope in a safe configuration

    • Load your sample

    • Find and adjust the focus

    Once completed, your sample will be ready for acquisition.

    UI Expand
    titleShutdown
    1. Save your data
    2. Close NIS-Elements software
    3. Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
    4. Turn off the computer
    5. If the oil objective was used, clean it with lens cleaner and lens paper (not Kimwipes)
    6. Wait until the Spectra has cooled down and turn off the microscope power bar (#2)
    7. Cover the microscope
    Note
    titleImportant Reminders
    • Collect your samples, especially those in the microscope
    • Leave the microscope and workspace clean


    UI Expand
    titleStorage management
    • Files can be saved temporarily (during acquisition) to local C: drive (desktop)
    • At the end of each session, copy your data to your external drive and delete it from local C: drive
    • You can store your files on the D: drive (Data Storage). If you do, please create one folder per laboratory using the principal investigator's last name. Inside, create a folder per user using the following nomenclature (First Name_Last Name).
    Note
    titleImportant

    In any case, do not store your files on the C: drive.



    UI Expand
    titleFirst use

    Anchor
    FirstUse
    FirstUse
    When using the microscope for the first time, you need to import the microscope settings into the software. You will usually do this during the training session.
    This procedure can also be performed if something is not working properly and if you want to reset the software to its original settings.

    Note

    This process will delete all experiment protocols and reset the software to the original settings for this specific microscope.

    1. If open, close NIS-Elements and wait until it is completely closed (up to 30 seconds)
    2. On your Desktop open the Softwares folder
    3. Open NIS Settings Utility
    4. Click on the Import tab
    5. Click on Browse
    6. Navigate to your Desktop
    7. Select the file Nikon Ti2-Fura settings for NIS.bin
    8. Click Select
    9. Select all items
    10. Click Import
    11. Click OK
    12. Close the NIS Settings Utility
    13. You can now reopen NIS-Elements



    Tabs Page
    idManuals
    titleManuals


    Available manuals


    Tabs Page
    idLog
    titleLog


    UI Expand
    expandedtrue
    titleTo do
    • Fix camera adapter
    • Purchase 3mm adapter for Lapp
    • Purchase multiband pass cube for Spectra
    • Fix stage inserts
    • Add Stage insert screws
    UI Expand
    title2025-03-10 Insallation Desmarais 2236
    • Complete installation
    • Complete cleaning


    Tabs Page
    idTechnical Datasheet
    titleTechnical Datasheet

    Stand

    • Nikon Ti2-E inverted  Serial System

    Light sources

    • Transmitted LED light
      • ND filter:  TBD
    • CoolLED pE340-Fura Serial
    • Lumencor Spectra Serial Serial

    Condenser

    • Manual condenser Serial

    • Lens LWD NA 0.52

    • Filter turret 7 manual positions

      1. ND TBD

      2. Empty
      3. Empty
      4. Empty
      5. Empty

    Objectives

    • 4x/0.2 Air WD 20
    • 10x
    • 20x/0.75 Air DIC WD 1.0
    • 40x
    • 60x/1.4 Oil DIC WD 0.13

    Stage

    • Motorized stage Encoded  Serial
    • Remote control joystick Ti2-S-JS Serial 
    • Inserts
      • Combo slide 3cm dish Ti2-S-HU with tilt adjustment (no incubation)

    Filters

    1. DAPI Cube TBD
    2. GFP Cube TBD
    3. Cy3 Cube TBD
    4. Cy5 Cube TBD

    Detector

    • Photometric Prime 25mm CMOS Monochrome Camera x 2048 pixels, 16-bit, 30fps at full resolution Serial 

    Workstation

    • HP Z440 Workstation
    • Intel Xeon E5-1620 v4 @ 3.5GHz
    • RAM 32 GB DDR4 2400 MHz ECC (4 x 8 GB)
    • OS 500 GB SSD 530 MB/s
    • 4 TB HD Data Storage (2 x 2 TB spanned volume) 130 MB/s
    • Video Card nVidia GTX 1080 8GB DDR5 dedicated memory
    • Monitor HP Z24i display 24' 1920x1200
    • Software NIS-Elements AR v5.02

    Consumables

    • Liquid Light Guide


    Tabs Page
    idFAQ
    titleTroubleshooting & FAQ


    Troubleshooting

    UI Expand
    titleCamera Driver Failed

    This can happen when the acquisition software is turned on before the camera has fully initialized.

    • Turn off NIS-Elements
    • Ensure the Initialize light at the back of the camera no longer blinks
    • Start NIS-Elements

    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 Container
    idDemo Image
    titleNikon Ti2-E
    directionhorizontal
    Tabs Page
    idDemo Image
    titleDemo Image

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