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Nikon TE2000-U inverted microscope

Bombardier Building, Room 3115

Instrument awarded to Dr. Luc Desgroseillers by the Canadian Foundation for Innovation (CFI)

  • Transmitted light
  • Phase Contrast
  • Fluorescence
  • Widefield imaging

Light sources

  • Halogen lamp for transmitted light

  • Lumencor Sola V-nIR for fluorescence

Emission (nm)

Nominal power (mW)

Objectives

  1. 4x/0.1 Air
  2. 20x/0.40 Ph1 Air
  3. 40x/0.75 Ph2 Air
PositionNameBrandFull nameIdentifierMagnificationNumerical apertureImmersionWorking distance (mm)Transmittance
(% [nm])		ApplicationsCover glass thickness (mm)
14x/0.10 AirNikon4x/0.1 PlanMRP000424x0.1Air30>80% [390-900]BF, Fluo-
220x/0.40 Ph1 AirNikon20x/0.40 LWD  Ph1 ADLMRP2620220x0.4Air3>80% [420-800]BF, PhC, Fluo1.2 !
340x/0.75 Ph2 AirNikon40x/0.75 Plan Fluor Ph2 DLLMRH1040540x0.75Air0.72>80% [400-700]BF, PhC, Fluo0.17
4Empty






  

5Empty






 

6Empty









BF: Bright-field
PhC: Phase Contrast
DIC: Differential Interference Contrast
Fluo: Fluorescence

Filters

  1. Empty

  2. UV: DAPI

  3. B: FITC

  4. G: Cy3

Filter cubes

PositionNameBrandIdentifierExcitationDichroicEmission
1Empty









2DAPIChroma

31000 v2

AT350/50x

400dclp

D460/50m

3FITCChroma

41001

HQ480/40x

Q505lp

HQ535/50m

4Cy3Chroma41007

HQ535/50x

Q565lp

HQ610/75m

5Empty




6Empty




Detector

  • CoolSnap HQ2 CCD

    • Monochrome Camera

    • 1392 x 1040 pixels

    • Pixel  6.5 um x 6.5 um

    • 10 f/s at full frame

    • Dynamic range 1 : 4000 12-bit 

    • Spectral response >50% between 410-680nm, Max 60% between 489-600nm

  1. If not already done, turn on the computer (#1)
  2. Remove the cover from the microscope
  3. Turn on the microscope power bar (#2)
  4. Start Metamorph

This procedure puts the microscope in a safe configuration to load your sample. At the end the microscope will be ready for acquisition.

  1. If not already done, select the 4x objective

    The 4x objective is the safest because it has the longest working distance (30mm). The sample will appear perfectly sharp long before the lens approaches it. It is recommended to always first focus with the safest lens.

  2. Place the test slide on the microscope stage with the coverslip toward the objective

    Important

    Always use the test slide to perform the first focus.

  3. If necessary, move the stage so that the sample is centered on the objective

For brightfield

  1. Turn on the transmitted light and adjust the intensity (on the left of the microscope)
  2. Select the first filter cube (double arrow)
  3. Set the lightpath selector to 1 (ocular) (on the right of the microscope)
  4. Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp

For fluorescence

  1. Press Mode on the fluorescence light remote to turn it on
  2. Adjust the intensity with the knob
  3. Select the desired filter cube UV: DAPI, B: FITC, or G: Cy3
  4. Set the lightpath selector to 1 (ocular) (on the right of the microscope)
  5. Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp



Important

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

  1. Select the 20x or 40x objective. The 40x objective is the best Air objective because it has the largest numerical aperture (0.75).
  2. Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp 

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).

Important

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

  1. Save your data
  2. Select the lowest magnification objective
  3. Adjust the focus to place the objectives in their lowest position
  4. Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
  5. Turn off the computer
  6. Turn off the microscope power bar (#2)
  7. Cover the microscope

Important Reminders

  • Collect your samples, especially those in the microscope
  • Leave the microscope and workspace clean

  • Complete installation
  • Complete cleaning

Stand

Nikon TE2000-U inverted

Light sources

Transmitted Halogen light Nikon TE2-PS100W Serial 12V

  • GIF Filter
  • Diffusion
  • Infra-red
  • Neutral color

Lumencor Sola nViR

  • EXFO Collimator 810-00030
  • Nikon Manual Neutral density filters ND8 ND4
  • Manual Fluorescence field diaphragm
  • Manual Fluorescence shutter

Condenser

  • Manual condenser
  • Lens LWD NA 0.52
  • Filter turret 5 manual positions

    • A BF

    • Ph1
    • Ph2
    • Empty
    • Empty

Objectives

See description

Stage

  • Manual Stage
  • Circular insert

Filters

See description

Detector

Roper Scientific Coolsnap HQ2
CCD image sensor Sony ICX285
CCD format 1392 x 1040
6.45 x 6.45-μm pixels
8.98 x 6.71-mm imaging area (optically centered)
System gain 1 e-/ADU
Linear full well 16,000 e- 
Read noise 4.5 e- rms @ 10 MHz 5.5 e- rms @ 20 MHz
Nonlinearity <1%
Digitizer type IEEE-1394a LVDS 14 bits @ 20 MHz or 10 MHz (software selectable)
Frame readout 90 ms/frame
CCD temperature -30°C (regulated)
Dark current 0.001 e-/p/s @ -30˚C

Workstation

TBD

Consumables

Liquid Light Guide
12V halogen bulb

Troubleshooting

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

  • Turn off Metamorph
  • Turn off the camera
  • Wait few seconds
  • Turn the camera back on
  • Wait until the camera has fully initialize
  • Start Metamorph

FAQ

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.

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Content published under CC BY-SA 4.0. Sharing allowed under the same license with attribution: "Original content by l'Institut Courtois d'innovation biomédicale, used under CC BY-SA 4.0"