Spectral response >50% between 410-680nm, Max 60% between 489-600nm
If not already done, turn on the computer (#1)
Remove the cover from the microscope
Turn on the microscope power bar (#2)
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.
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.
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
For brightfield
Turn on the transmitted light and adjust the intensity (on the left of the microscope)
Select the first filter cube (double arrow)
Set the lightpath selector to 1 (ocular) (on the right of the microscope)
Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
For fluorescence
Press Mode on the fluorescence light remote to turn it on
Adjust the intensity with the knob
Select the desired filter cube UV: DAPI, B: FITC, or G: Cy3
Set the lightpath selector to 1 (ocular) (on the right of the microscope)
Adjust the focus with the main dial while looking through the eyepieces until the image is perfectly sharp
Important
First focus with thesafest objective before selecting another lens and continuing with secondary focus.
Select the 20x or 40x objective. The 40x objective is the best Air objective because it has the largest numerical aperture (0.75).
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.
Save your data
Select the lowest magnification objective
Adjust the focus to place the objectives in their lowest position
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 microscope power bar (#2)
Cover the microscope
Important Reminders
Collect your samples, especially those in the microscope
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.
