Flow Cytometry

Researcher preparing samples for the flow cytometer.

The Flow Cytometry Core’s primary function is to offer Center Investigators the ability to utilize complex and technically-challenging flow cytometric approaches to characterize cells and their functional and signaling properties. The Core is also committed to an educational goal. Not only will Center members be trained on the operation of instruments, it is the intent that all users completely understand cytometric theory, application, and data analysis.


  1. Support current COBRE and Center projects. The Core provides state-of-the-art equipment and the technical and analytical expertise to address the flow cytometric needs of the supported early career investigators and other members of the Diabetes and Obesity Center at the University of Louisville.
  2. Educate and train investigators. Many of our Cores organize seminars and informal meetings to serve as a forum in bringing together users and to initiate discussion of novel reagents and applications. It is intended that all COBRE-supported early career investigators, in their transition to independence, will completely understand cytometric theory, application, and data analysis and be able to competently incorporate flow cytometry as an analytical tool in their future studies.
  3. Develop new protocols and techniques. The Core is currently expanding its operations to support the multidimensional flow cytometry needs in the future. With the incorporation of new investigators into the Center in the future,new projects and experiments will be carried out to perform cutting-edge research to address the rapidlyspreading epidemics of diabetes and obesity.

James McCracken, PhD at work in the flow cytometry laboratory

The Core is housed in a 400 sq. ft. laboratory in the Baxter II building at the Health Science Center at the University of Louisville. This lab is fully equipped with the LSR II and Acuri C6 cytometers (described below) centrifuges, computers, refrigerators and freezers, tissue culture hood for sterile techniques, incubators, and all the small equipment needed for cell handling and staining. The MoFlo cell sorter (described below) is housed in the adjacent MDR building.

MoFlo: Housed in the MDR building, this is the DOC’s cell sorter. Currently set up for 5 color, 2 laser detection, it can sort cells at rates up to 40,000 events/second. This instrument is currently only available by reservation with the Technical Director as it requires a dedicated, trained operator.

BD-LSRII: This Becton Dickinson (San Jose, CA.) instrument is housed within the cytometry room (Baxter II Room 207). It is equipped with 3 solid-state lasers; a 488 nm (Coherent), 633 nm (JDS Uniphase), and 405 nm (Coherent). The LSRII has recently been upgraded with appropriate detectors, mirrors and filters for up to 11 color detection (5 off the 488 laser, and 3 each off the 633 and 405 lasers).

BDAccuri C6: This is another Becton Dickinson instrument housed in Baxter II. It has two lasers (488nm and 640nm) and 4 color-detectability. A unique feature of this instrument is its cell counting capability.

Abbott Cel-Dyn 3200: This instrument is used for the complete blood counting (CBC) of human and animal (mouse, rat, rabbit, non-human primate) samples.

Fee for analyzer use

Investigators

Source of funding

Reagents

Hourly fee

Individual projects

NA

Core supplied

none

Center members

any

User supplied

$40

Non-Center users

federal

User supplied

$57

Non-Center users

Any

User Supplied

$60

Non-U L users

 

 

$100

Fee for operator time (processing, acquisition, analysis}

Investigators

Source of funding

Hourly fee

Individual projects

NA

none

Center members

any

$20

Non-Center users

federal

$27

Non-Center users

Any

$30

Non-U L users

 

$60


CBC use: $10/sample, free to Individual Projects
Sorting to be billed as Instrument time+operator. 1 hour operator time for set up and shut down for each session is required.


Contact

Email James McCracken
580 S. Preston St.
Baxter II, Room 207
Louisville, KY 40202


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