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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-E fully motorized inverted microscope

Roger Gaudry R-619

  • Applications

    • Brightfield

    • Phase contrast

    • Polarized light, DIC

    • Fluorescence

    • Live imaging
    • Long timelapse imaging

  • Light sources

    • LED lamp for transmitted light

    • Lumencor SpectraX for fluorescence 

Développer
titleLumencor SpectraX complete specifications


Filter nameFilter IDFilter typeExcitation wavelengths (nm)Compatible fluorophores
V

395/25

Bandpass[382-407]

DAPI, Hoechst

B

440/20

Bandpass[430-450]CFP
G

470/24

Bandpass[458-482]FITC, GFP
NT

510/25

Bandpass[497-522]YFP
GY550/15Bandpass

[542-557]

TRITC, Cy3

GY (Storage)

575/25Bandpass[562-587]

mCherry

R

640/30

Bandpass[625-655]

Cy5


  • Objectives

    • 20x/0.5 Air Ph1
    • 60x/1.4 Oil DIC WD 0.13
    • 100x/1.45 Oil Ph3 WD 0.13
    • 100x/1.45 Oil DIC WD 0.13
    • Empty
    • 20x\/0.75 Air DIC 
Développer
titleObjectives complete specifications


PositionNameBrandIDMagnificationNumerical ApertureImmersionTypeWorking distance (mm)Transmittance
(% [nm])		TechniqueCover glass thickness (mm)
120x/0.5 Air Ph1Nikon

20x/0.5 Air Plan Fluor Ph1

20x0.5AirPlan Fluor2.1>80% [400-750]BF, Pol, PhC, Fluo0.17
2

60x/1.4 Oil DIC

Nikon60x/1.4 Oil Plan Apo Lambda DIC N260x

1.4


Remarque
Oil


Plan Apo Lambda0.13>80% [475-725]BF, Pol, DIC, Fluo0.17
3

100x/1.45 Oil Ph3

Nikon100x/1.45 Oil Plan Apo Lambda Ph3

100x

1.45


Remarque
Oil


Plan Apo Lambda0.13>80% [475-750]BF, Pol, PhC, Fluo0.17
4100x/1.45 Oil DICNikon100x/1.45 Oil Plan Apo Lambda DIC N2

100x

1.45


Remarque
Oil


Plan Apo Lambda0.13>80% [475-750]BF, Pol, DIC, Fluo0.17
5Empty









620x/0.75 Air DICNikon20x/0.75 Air Plan Apo Lambda DIC N220x0.75AirPlan Apo Lambda

1.0

>80% [400-950]BF, Pol, DIC, Fluo0.17


  • Filter cubes

    1. DAPI/Hoechst/AMCA

    cubes

    1. cube

    2. FITC/EGFP

    cubw

    1. cube

    2. TRITC/Rhodamine

    3. DAPI/FITC/TRITC/Cy5

    4. DIC Analyser
  • CFP\YFP\mCherry

6 positions motorized

  • Ex 383-408 DAPI-U DM 425 BA 435-485
  • Semrock 96372 M349727 17
  • Semrock 96376 M351081 8
  • DAPI/FITC/TRITC/Cy5 77074160 Custom Quad C182279 Polychroic and quad bandpassing single bandpass filters: ET395/25x, ET470/24x, ET550/15x, ET640/30x CFP\YFP\mCherry7707\4656
  • Ti2CDICACL
    1. CFP\YFP\mCherry
    XT C197767
Développer
titleFilters complete specifications


PositionCube nameBrandIDExcitation FilterDichroic mirrorEmission Filter
1DAPI/Hoechst/AMCAChromaCube set 31000v2350/50x [325-375]400LP460/50m [435-485]
2FITC/EGFPChromaCube set 41001

480/40x [420-500]

505LP

535/50m [510-560]
3TRITC/RhodamineChroma

Cube set 41002c

545/30x [530-560]570LP620/60m [590-650]
4Texas Red
  1. DAPI/FITC/TRITC/Cy5
Chroma

Cube set 41004

560/55x [532-588]

595LP645/75m [607-682]
5Empty
  1. DIC Analyser





6Empty
  1. CFP\YFP\mCherry






  • Detector


Tabs Page
idUser Guide
titleUser Guide


UI Expand
expandedtrue
titleStartup
  1. Turn on the computer
  2. Turn on the microscope power bar

  3. If incubation is required, turn on the Okolab incubation module (3A and 3B) and open the CO2 (3C) and N2 (3D) tanks

    Avertissement

    Make sure the water bath is clean and properly filled with distilled water


  4. Log in Windows using your UdM credentials
  5. Start-up NIS-Elements
Info

The first time you log in the computer, you need to import the microscope settings. To do this follow the instructions Setting-up NIS-Elements



UI Expand
titleShutdown
  1. Save your data
  2. Close NIS-Elements
  3. Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
  4. Get your samples from the microscope
  5. Clean the oil objectives with lens cleaner and lens paper
  6. If incubation was used, turn off the incubation module (3A and 3B) and close the CO2 (3C) and N2 (3D) tanks
  7. Turn off the microscope power bar (2)
  8. Cover the microscope
Remarque
titleImportant Reminders
  • Take back your samples including ones in the microscope
  • Leave the microscope and the working area clean



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
titleSetting-up NIS-Elements

Ancre
Setting-up NIS-Elements
Setting-up NIS-Elements

This process is required the first time you are using the instrument. You will usually do it during the training session. It can also be performed if something is not working properly right or if you want to refresh the software interface.

Remarque

This process will delete all experiment protocols and restore the parameters for the microscope.

  1. Close NIS-Elements
  2. Wait until the complete closure of NIS-Elements
  3. Open the folder Desktop\Logiciels
  4. Open the software NIS Settings Utility
  5. Click on the Import tab
  6. Click on Browse
  7. Navigate to your desktop
  8. Select the file Nikon-Ti2 Settings.bin
  9. Click Select
  10. Select all items
  11. Click Import
  12. Click OK
  13. Close the NIS Settings
  14. Open NIS-Elements



Tabs Page
idLightpath
titleLight path

Light path

Download the schematics to follow Transmitted light (Bright field, Phase Contrast, Polarized light, DIC) and Reflected light (fluorescence) paths.

Light path schematics.pdf


Tabs Page
idManuals
titleManuals

Manuals


Tabs Page
idLog
titleLog


UI Expand
expandedtrue
titleTo do
  • Add label on gas bottle
  • Add line mark on humidifer
  • Adjust focus drive jitter
  • Add a holder for polarized light analyzer


UI Expand
title2018-04-04
  • Nikon Preventive Maintenance
  • Adjustment and cleaning Stage Nosepiece, condenser, filter turret, focus drive, shutters
  • 100x slightly damaged but not the lens
  • Fluorescence cubes damaged: FITC TexasRed
  • Focus drive has a jitter
UI Expand
title2021-07-20
  • Replacement mercury lamp bulb HBO 1003W/2
  • 256 512 GB SSD added in workstation installed for OS
  • Windows 10 Installation
  • BIOS updated to v2.47
  • Camera Firmware updated to v2.11
  • NIS-Elements Basic Research v4.6 64-bits installed
  • Creation NIS Elements Settings
  • FITC and TxRed cubes excitation and emission filters slightly damaged. Dichroic OK



570LP

Detector

  • Hamamatsu ORCA Flash V2 C11440-22CU CMOS Monochrome Camera 2048 x 2048 pixels, 16-bit, 30fps at full frame Serial 101 081

Workstation

  • HP Z440 Workstation
  • Intel Xeon E5-1630 v3 1620 v4 @ 3.7GHz5GHz
  • RAM 32 GB DDR4 2133 MHz  2400 MHz ECC (4 x 8 GB)
  • OS 256GB 500GB SSD 550 MBsMB/s
  • 2TB 4TB HD Data Storage 150 MBs(2 x 2 TB spanned volume) 170 MB/s
  • Video Card NVIDIA Quadro K620 2 GB DDR3 nVidia GTX 1080 8GB DDR5 dedicated memory
  • Monitor HP Z24i display 24' 1920x1200
  • Software NIS-Elements AR v5.02

Incubation

  • Okolab BoldLine Temperature unit Serial 284-1058 H101 T Unit BL
  • Okolab BoldLine CO2/O2 Unit  0-10/1-18 Serial 088-1102
  • Okolab OkoTouch Serial 118-224

Consumables

  • CO2 Tank
  • N2 Tank
Tabs Page
idTechnical Datasheet
titleTechnical Datasheet

Stand

  • Nikon Ti2-E inverted  Serial 540156 System 170110-Sys-006287

Light sources

  • Transmitted LED light
    • ND32 filter
    • Manual Polarizer
  • Lumencor SpectraX 6-NII-SE Serial 9409 Filters B G R NT V

  • R 640\30-25

  • G 470\24-25
  • B 440\20-25
    • NT 510\25-25
  • V 395\25-25
  • GY : 550\15-25
  • Storage GY 575\25-25

Condenser

  • Motorized condenser

  • Lens LWD NA 0.52

  • Filter turret 7 motorized positions

    1. Empty

    2. Empty

    3. Ph1

    4. Ph2

    5. Shutter

    6. Empty

    7. DIC N1

Objectives

  • 10x20x/0.3 Plan Fluor Ph1 DLL WD 16 Air40x/0.75 Plan Fluor Ph2 DLL WD 0.72 Air with PF40 Wollaston prism5 Air Ph1
  • 60x/1.4 Oil DIC WD 0.13
  • 100x/1.3 Plan Fluor 45 Oil Ph3 DLL WD 0.2 Oil with PF/PA 100 Oil Wollaston prism
  • Empty
  • Empty
  • Empty
  • 13
  • 100x/1.45 Oil DIC WD 0.13
  • Empty
  • 20x/0.75 Air DIC
Développer
titleObjectives complete specifications


PositionNameBrandIDMagnificationNumerical ApertureImmersionTypeWorking distance (mm)Transmittance
(% [nm])		TechniqueCover glass thickness (mm)
110x20x/0.3 5 Air Ph1Nikon10x

20x/0.

3

5 Air Plan Fluor Ph1

DLL

10x20x0.35AirPlan Fluor162.1>75% >80% [400-800750]BF, Pol, PhC, Fluo0.17
2

40x60x/0.75 Air1.4 Oil DIC

Nikon40x60x/01.75 Air Plan Fluor Ph2 DLL40x

0.75

Air

Plan Fluor0.724 Oil Plan Apo Lambda DIC N260x

1.4


Remarque
Oil


Plan Apo Lambda0.13>80% [475-725>80% [400-750]BF, Pol, PhCDIC, Fluo0.17
3

100x/1.345 Oil Ph3

Nikon100x100x/1.3 45 Oil Plan Fluor Apo Lambda Ph3 DLL

100x

1.345


Remarque
Oil


Plan FluorPlan Apo Lambda0.213>75% >80% [400475-800750]BF, Pol, PhC, Fluo0.17
4Empty100x/1.45 Oil DICNikon100x/1.45 Oil Plan Apo Lambda DIC N2

100x

1.45


Remarque
Oil


Plan Apo Lambda0.13>80% [475-750]BF, Pol, DIC, Fluo0.17
5Empty









6Empty20x/0.75 Air DICNikon20x/0.75 Air Plan Apo Lambda DIC N220x0.75AirPlan Apo Lambda

1.0

>80% [400-950]BF, Pol, DIC, Fluo0.17


Stage

  • Motorized stage Ti2 SHU compatible Serial 127808
  • Remote control joystick Ti2-S-JS Serial 127976
  • Inserts
    • Combo slide and 3cm dish with tilt adjustment insert
    • Multiwell Multi-well plate insert

Filters

  • Ex 383-408 DAPI-U DM 425 BA 435-485
  • Semrock 96372 M349727 17
  • Semrock 96376 M351081 8
  • 77074160 Custom Quad C182279 Polychroic and quad bandpass emitter for use with the following single bandpass filters: ET395/25x, ET470/24x, ET550/15x, ET640/30x
  • DIC Analyser Ti2CDICACL
  • 7707\4656 CFP\YFP\mCherry XT C197767
Développer
titleFilters complete specifications
PositionCube nameBrandIDExcitation FilterDichroic mirrorEmission Filter
1DAPI/Hoechst/AMCAChromaCube set 31000v2350/50x [325-375]400LP460/50m [435-485]
2FITC/EGFPChromaCube set 41001

480/40x [420-500]

505LP

535/50m [510-560]
3TRITC/RhodamineChroma

Cube set 41002c

545/30x [530-560]620/60m [590-650]
4Texas RedChroma

Cube set 41004

560/55x [532-588]

595LP645/75m [607-682]
5Empty6Empty


Tabs Page
idFAQ
titleTroubleshooting & FAQ


UI Expand
expandedtrue
titleTroubleshooting

Liquid forming in the incubation chamber

This happens when the mixed-gas humidifier is overfilled. Bubbles created by the gas going through the humidification bring liquid into the gas feed line.

  • Turn off the Okolab module
  • Remove your sample and store it properly
  • Dry the incubation chamber with a clean tissue
  • Carefully remove the cap of the humidifier glass bottle
Avertissement

The humidifier bottle is made of glass and is very fragile. Pay extra care when manipulating the humidifier bottle

  • Remove distilled water from the humidifier
Info

Humidifier shouldn't be more than 2/3rd filled

  • Close the humidifier by replacing the cap
  • Turn on the Okolab module


UI Expand
titleFAQ

Can 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 as well as slides between a glass
  • 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 cover slip.coverslip





Tabs Container
idDemo Image
directionhorizontal


Tabs Page
idDemo Images
titleDemo Images