Flow Cytometry

Director: Jun Yan, M.D., Ph.D.

Manager: Christopher A. Worth, Biomedical Engineer Sr.

Contact:  Chris  A. Worth: 852-7191   Chris.Worth@louisville.edu

The online scheduling system can be accessed at this link.

Introduction

This facility traces its roots back to 1991, with the purchase of a Coulter Elite Cell Sorter In March of this year the facility was moved to CTR 340.  In 2014 the system was upgraded to the MoFlo XDP system.  We are seeing increased sorting efficiency and speed with the new electronics. Additionally, the MoFlo’s CyClone system is able to sort into a larger range of tissue culture plates including 6,12,24,96 and Terasaki plates, for example.  The CyClone is capable of having different sort criteria for each well.

Capabilities

The MoFlo includes:

  • 11 parameter, 9 color, 4 way sorting.
  • 488nm Blue solid state LASER, providing FALS and SSC parameters. Five colors available to use.
  • 633nm Red solid state LASER, with Two colors available.
  • 405nm Violet solid state LASER, with two colors available.
  • The aforementioned Cyclone system for Cloning,  using standard plates, in addition, customized target plates can be programmed.
  • Sorted cells are typically collected on ice in tubes of various sizes, depending on user requirements.
  • Sorted cells are biologically viable for a wide range of additional experiments.

 

Example of Sorting Experiment

This facility did this project for Richard L. Benton, Ph.D., Dept. of Anatomical Sciences & Neurobiology, Kentucky Spinal Cord Injury Research Center (KSCIRC).  These experiments marked the first time these cells were ever isolated by FACS.  Also, it is rare for sort products to be photographed.  This gives the viewer a much better idea of a sort result instead of the typical histograms and dot plots.  The cells pictured were sorted from the R1 region in the dot plot.  We determined that the R2 population was debris.

Amnis ImageStream IS100

Think of it as a Flow Cytometer with a Microscope attached.  This system allows you to image each cell you send through the instrument, with up to four fluorochromes. Our first generation unit has a 488nm blue excitation LASER.  Enabling users to work with their familiar Flow Cytometry fluorochromes, FITC, PE, 7-AAD, PE-Cy5, PE-Cy5.5, PerCP, Draq-5

This system can be used for experiments such as

  • Translocation of signaling molecules
  • Co-localization of molecules
  • Internalization / phagocytosis
  • Apoptosis/necrosis/autophagy
  • Morphology-based cell classification

Aperio CS-GL

A touch of a button will scan a single slide Create a digital slide every 5 minutes (24bit True color with gigapixel resolutio)