Nikon TE2000
Long Term Live Cell Imaging

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Hardware:

  • Nikon TE2000 inverted microscope stand with DIC and phase contrast optics
  • Prior Proscan II motorized ZY stage and focus motor
  • Uniblitz fluorescence and transmitted light shutters
  • EXFO metal halide light source
  • Solent Scientific incubation enclosure
  • Hamamatsu Orca-ER cooled CCD camera
  • MetaMorph aquisition software

Filters:

Not sure which fluorophores will work best? Use an online spectra viewer (see Links) or contact Wendy.

Software setting Excitation Dichroic Emission
DAPI D360/40X   ET460/50M
GFP HQ480/40X   535/50M
RFP 560/40X   D630/60M
Cy5 628/40X   692/40M
DIC Tungsten halogen   Analyzer
Brightfield Tungsten halogen   Specified filtercube
*CFP ET436/20x T455lp ET480/40m
YFP ET500/20x T515lp ET535/30m
*CFP/YFP FRET ET436/20x T455lp ET535/30m
*BFP ET405/20x T425lpxr ET460/50m

All numbers in nanometers.
*Filters marked with an astrisk are not permanently installed. Contact Wendy for use of these.

Objectives:

Mag. NA

Immersion Medium

Phase contrast DIC Corrections **Scaling (um/pixel)
10x 0.30 Air Ph 1 -- Plan Fluor 0.647
20x ELWD 0.45 corr Air Ph1 -- Plan Fluor 0.328
40x ELWD 0.6 corr Air Ph2 -- Plan Fluor 0.166
*40x 0.95 Air -- H Plan Apo 0.164
*40x 1.3 Oil -- H Plan Apo  
*60x 1.4 Oil *Ph3 -- Plan Apo 0.108
*100x 1.4 Oil *Ph3 -- Plan Apo  

corr = objective has a correction collar that must be adjusted to prevent spherical aberration.
*Objectives marked with an astrisk are not permanently installed. Please contact Wendy for installation.
** These values work well for making scale bars. When performing careful distance measurements, it is best practice to confirm the calibration for the exact setup you use.

Sample holders:

All objectives, except ELWD, expect a #1.5 glass coverslip. ELWD can work with either glass coverslips or plastic dishes. Glass must be used when imaging fluorescence.

Stage plates Compatible Sample holders
Adjustable Slides, LabTek II chambers, 35mm dishes, 60mm dishes
Multipwell plate Any standard 96-well plate format

Software:

To view files in lab, many users like to use Fiji (ImageJ) with the BioFormats plugin for importing the .nd file. See our Links page to download.

 

 
 

 

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For website assistance please contact Nicki Watson