Instrument awarded to Dr. Steve Michnick by the Canadian Foundation for Innovation (CFI)
Transmitted light
Phase Contrast
Interference Contrast (DIC)
Fluorescence
Widefield imaging
Live imaging
Time-lapse imaging
Nikon TE2000EImage taken with the Nikon TE2000E
Nikon TE2000-E inverted microscope
Bombardier Building, Room 3132 Instrument awarded to Dr Steve Michnick by the Canadian Foundation for Innovation (CFI) Advanced Microscope Tier 1 usage price
Hamamatsu ORCA Flash v2 CMOS Monochrome Camera 2048x2048 pixels, 16-bit, 30 images/s at full frame (100 images/s at full frame with camera link connector). Spectral response >40% between 400-850nm. Max 80% at 550nm.
If not already done, turn on the computer (#1) and use your UdeM credentials to log in to Windows
Remove the cover from the microscope
Turn on the X-Cite exacte light source (#2)
Turn on the microscope power bar (#3)
When using the software for the first time, it is necessary to import the microscope-specific settings before launching the acquisition software. Please refer to the First Use section below for instructions.
Start NIS-Elements
When using the acquisition software 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.
Please note, this procedure will delete all your experiment protocols and restore the software to its original settings.
If open, close NIS-Elements and wait until it is completely closed
On the Desktop open the folder Softwares
Open NIS Settings Utility
Click on the Importtab
Click on Browse
Navigate to your Documents
Select the file NIS Settings for Nikon TE2000E.bin
Click Select
Select all items
Click Import
Click OK
Close NIS Settings Utility
You can then open NIS-Elements
This procedure puts the microscope in a safe configuration to load your sample. At the end the microscope will be ready for acquisition.
If not already done, in NIS-Elements select 4x to 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.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 15.2 mm.
If not done already, press Escape to lower the objective to the lowest position
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.
If necessary, move the stage so that the sample is centered on the objective
In NIS-Elements, select the optical configuration BF vis or the desired fluorescence (DAPI vis, ...) to activate the configuration
Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
The focus can be found at Z = mm). The Z value can be found in the NIS Software XYZ Position tab.
In NIS-Elements, select the optical configuration Off to turn off the illumination
Important
First focus with thesafest objective before selecting another lens and continuing with secondary focus.
In NIS-Elements, click 10x, 20x or 40x to select the desired objective
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).
In NIS-Elements, select the optical configuration BF vis or the desired fluorescence (DAPI vis, ...) to activate the configuration
Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
Select the optical configuration Off to turn off the illumination
Your sample is ready for acquisition!
After performing the first focus, in NIS-Elements:
In NIS, click Escape, to lower the objectives
Click 60x Oil or 100x Oil to select the desired objective.
The 100x objective has the greatest resolution (NA 1.45 vs 1.4)
Place a single drop of oil on the objective
In NIS-Elements, click again Escape to return the objective to its original position
Select the optical configuration BF vis or the desired fluorescence (DAPI vis, ...) to activate the configuration
Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
Select the optical configuration Off to turn off the illumination
Your sample is ready for acquisition!
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.
Save your data
If the oil objective was used, clean it with lens cleaner and lens paper (not Kimwipes)
Select the lowest magnification objective and press Escape to place the objectives in a safe position
Close NIS-Elements
Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
Turn off the computer
Turn off the X-Cite exacte light (#2)
Turn off the microscope power bar (#3)
Cover the microscope
Important Reminders
Collect your samples, especially those in the microscope
Perfect Focus avec LED de 780 nm et avec filtre dans le trajet lumineux LP 750 nm (inadéquat pour le Cy5.5)
En lumière transmise, pour que le PFS fonctionne, il faut insérer 2 filtres IR dans le trajet lumineux: celui du PFS et celui du statif près de la lampe halogène
Complete installation
Complete cleaning
Stand
Nikon TE2000-E inverted
Nikon T-HUBC Serial 750110
Nikon PFS T-PFS 810060
Nikon T-RCP Serial 750104
Light sources
Transmitted Halogen light Nikon TE2-PS100W Serial 505320 12V
GIF Filter
Diffusion
Infra-red
Neutral color
Transmitted light shutter uniblitz VCM D1 Serial A607295
X-Cite exacte Model XCT10A 200 W Seria;l XCT10A-0337
EXFO Collimator 810-00030
Prior Shutter HF201HT Serial 7332
Prior 8 position motorized excitation filter wheel 61817
Nikon Manual Neutral density filters ND8 ND4
Manual Fluorescence field diaphragm
Manual Fluorescence shutter
Condenser
Motorized condenser
Lens LWD NA 0.52
Filter turret 5 motorized positions
A BF
DIC M
Ph1
Ph2
Ph3
Objectives
See description
Stage
Prior Proscan 2 H30V4 Serial 62040
Remote control joystick Prior CS152v2 Serial R0772128
Inserts
1 slide
Multi-well plate
Adapter for 3cm dish
Filters
See description
Detector
Hamamtsu ORCA Flash V2 CMOS Model C11440-22CU Serial 002162 V2
Monochrome Camera 2048 x 2048 pixels
Pixel 6.5 um x 6.5 um
Force air cooled at -10C
Effective sensor area 13.312 mm (H) × 13.312 mm (V
Dark current 0.06 electron/pixel/s
Reading noise 1.6 electrons (r.m.s) at standard scan
Dynamic range 1 : 18 000
Spectral response >40% between 400-850nm. Max 80% at 550nm
