Cytometry & Imaging Microscopy Core Facility

Director: James W. Jacobberger, PhD
Professor, General Medical Sciences (Oncology)
jwj@case.edu


Cytometry Instrumentation

Cytometers are quantitative microscopes – the light emitted, reflected, or refracted from small things is imaged onto photomultiplier tubes, photo-diodes, avalanche diodes, or CCD camera and the amplified resulting electrical signal is converted to digital form and subsequently processed by the cytometer's hardware and software. The result is a list of data for each cell sampled from a population. Since all cell populations are heterogeneous or asynchronous (despite efforts to purify and synchronize), multiparametric cytometer data are correlated in a meaningful way within samples.

Cell Sorting

  • BD Biosciences Aria
    • 3 lasers; 9 color (blue, green, orange, red, far-red) emission
    • Compatible dyes: Hoechst 33342, DAPI, Pacific Blue, Pacific Orange, Fluorescein/A488, GFP, PI, AO, 7AAD, PE, A555, Cy3, YFP, PE-Cy5, PE-Cy5.5, PE-Cy7,PerCp, PerCp-Cy7, A647, A700, AP, AP-Cy7
    • Four way sorting; multi-well plate sorting. Sort speed 25,000/s
  • ICyte Reflection
    • 5 lasers (choice of 2); 4 color (blue, green, orange, red, far-red) emission
    • Compatible dyes: Hoechst 33342, DAPI, Pacific Blue, Pacific Orange, Fluorescein/A488, GFP, PI, AO, 7AAD, PE, A555, Cy3, YFP, PE-Cy5, PE-Cy5.5, PE-Cy7, PerCp, PerCp-Cy7, Texas Red, RFP, A647, A700, AP, AP-Cy7
    • Four way sorting. Flexible, high speed. Sort speed 50,000/s

Flow Cytometry

  • Millipore Guava
    • 1 blue laser; 2 color (green and red) emission
    • Compatible dyes: Fluorescein/A488, PI, 7AAD
    • This cytometer is mainly used for live/dead cell counting and data are output as events per ml. Does not have light scatter parameters.
  • Beckman Coulter XL
    • 1 blue laser; 4 color (green, orange, red, far red) emission
    • Compatible dyes: Fluorescein/A488, GFP, PI, AO, 7AAD, PE, A555, Cy3, YFP, PE-Cy5, PE-Cy5.5, PE-Cy7, PerCp, PerCp-Cy7
  • BD Biosciences LSR 2
    • 5 lasers; 14 color (blue, green, orange, red, far red) emission
    • Compatible dyes: Hoechst 33342, DAPI, A350, Pacific Blue, Pacific Orange, Fluorescein/A488, GFP, PI, AO, 7AAD, PE, A555, Cy3, YFP, PE-Cy5, PE-Cy5.5, PE-Cy7, PerCp, PerCp-Cy7, A647, A700, AP, AP-Cy7

Imaging Cytometry

  • Compucyte iCyte laser scanning cytometer (LSC)
    • 3 lasers; 8 color (blue, green, orange, red) emission
    • Compatible dyes: Hoechst 33342, DAPI, Pacific Blue, Pacific Orange, Fluorescein/A488, GFP, PI, AO, 7AAD, PE, A555, Cy3, YFP, PE-Cy5, PerCp, A647, AP
    • This platform accepts chamber slides, standard culture dishes with coverslipped windows (Labtek); optical grade multiwall plates
  • Amnis Image Stream flow cytometer
    • 3 lasers; X color (blue, green, orange, red) emission
    • Compatible dyes: Hoechst 33342, DAPI, Pacific Blue, Pacific Orange, Fluorescein/A488, GFP, PI, AO, 7AAD, PE, A555, Cy3, YFP, PE-Cy5, PE-Cy5.5, PE-Cy7, PerCp, PerCp-Cy7, A647, AP