Evos                               (first draft of webpage)
ACLSW 421

Sign up online https://nyumc.ilab.agilent.com/account/login

Always sign in the paper logbook in the room too!

Want to read the whole manual? Here it is.

All users must be trained by Microscopy Laboratory staff. No more intralab training!!
New person in your lab needs to use Evos? You may show them what they need to do, but they may not use the system unassisted until checked out by Microscopy Lab staff.

Instrument calibrated for samples that have coverslips on them. Images from thicker bottom dishes may look ok at 10X and 4X, but lenses really designed for #1.5 coverslips.

One of the selling points of the Evos is the ability to use in a fully lighted room.  If you need to keep the sample dark, put the Evos supplied box over the sample or put black cloth or cardboard over sample.  Other people in the room may need the lights on and this is acceptable.

Please always put stage plates in the plastic bin with the other stage plates. Do not leave stage plates on table etc.


Instrument must be left in standard configuration with buttons in this order:
GFP
RFP
Cy5
Brightfield
Phase

There are Dapi & YFP blocks available. Please contact Microscopy Lab staff if you need to use them. (Most YFP imaging will work ok with the GFP block.)

 

WARNING: Images may not be saved with proper (or any) scaling information embedded in them.
Always put lens magnification in filename &/or folder name!

Based on measurements 20170308, each pixel with each lens is: (We will confirm these numbers and repost any changes if needed.)

Magnification N.A. XY color camera

YX grayscale camera

4X   2.25 um um
10X   0.9 um 0.9 um
20X   0.45 um um
40X   0.22 um um

If need a scalebar in ImageJ, Image > Properties and type in the X and Y for Pixel width and Pixel height respectively and then Analyze > Tools > Scalebar...

 

DO NOT TOUCH CALIBRATION SETTINGS IN TAB ON FAR RIGHT!!
When microscope calibrated correctly (as of today 8 Mar 2017), you can focus at 4X (image at left) and then click on the 40X lens and be in the same location needing only a tweak of the focus (image at right).


4X lens

40X lens (original size image 5X pixels linearly in each X and Y)

 

What about the corners being dark and slight gradient from top to bottom? We'll check with Evos, but these are probably unavoidable. Unless you crop them off. Or postprocess using flatfield correction.

 

 

When there is a problem, questions, suggestions, etc, please tell us michael.cammer@med.nyu.edu

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