Resources

Instructions

How to use FlowJo off-campus using an Emory VPN connection

Follow these instructions to create and open a VPN (Virtual Private Network) connection to the Emory system.

http://it.emory.edu/security/vpn.html

Then fire up FlowJo (make sure you've registered FlowJo!) Let us know if you have any questions.

How to connect to the Flow Core server for data transfer

Protocols and Procedures

Training Materials

Sorting

In order to avoid the common pitfalls inevitable in preparing cells for sorting, we recommend reading the following links:

Sorting Presentations

  • Source Lab PI: Dr. Rafi Ahmed, PhD
  • Client: Dr. Kim Kawamura, PhD
  • Source material: P14 chimeras (B6 mice initially transferred with 106 P14 splenocytes) and then immunized with DNA
  • Sort criteria: separate out CD62L hi and lo populations from DNA- immunized mice 62 days post-vaccination
  • Sort goal: to use the sorted cells to make RNA for gene chip analysis
  • Sort protocol: high-speed two-step sort

References

1. Flow Cytometry: A Practical Approach. Edited by MG Ormerod. ( IRL Press, Oxford. 1994. ISBN 0-19 963461-0)

2. Practical Flow Cytometry. 3rd Edition. Howard M Shapiro. ( Alan R Liss, Inc. ISBN 0-471-30376-3).

3. Flow Cytometry. First Principles. Alice Longobardi Givan. ( Wiley-Liss, New York, 1992. ISBN 0-471-56095-2.)

4. Handbook of Flow Cytometry Methods. Edited by J Paul Robinson. ( Wiley-Liss, New York, 1993. ISBN 0-471-59634-5. )

Facility Description

Detailed facility information is given below. If you need additional information please contact the Core Director.

Instrumentation

The Emory University School of Medicine Core Facility for Flow Cytometry is located in the Dental School Building.  Support in the form of training, technical assistance, troubleshooting and consultation are given to various laboratories that independently own cytometers on campus. Facility hardware:

All instruments have been configured with 5 lasers and 17 detectors.  The laser wavelengths are 355nm (UV), 407nm (violet), 488nm (blue), 561nm (yellow-green), 633nm (red).
 
Analyzer:
  • LSR II (Location: Dental School, Room 440A)
Sorters:
  • FACSAria IIA (Location: Dental School, room 441)
  • FACSAria IIB (Location:  Dental School, room 441)

The LSR II and FACSAria II are manufactured by Becton Dickinson.  Our sorters have capabilities of sorting up to 4 populations at one time and of single-cell deposition sorting, and all sorts are treated with great care in regards to aseptic techniques, accuracy in post-sort recovery and high-purity (>98%).

Personnel

Robert Karaffa, Technical Director (rkaraff@emory.edu)