Flow Cytometry: Using the Facility

The Biomedical Sciences Flow Cytometry Core Laboratory at Cornell University located in Ithaca NY provides a wide variety of cytometry services for investigators at Cornell University as well as external investigators. We currently have access to a hi-speed FACSAria cell sorter and an LSR II. We will be happy to help you establish or convert your assay to flow cytometry, as well as aid in the analysis and interpretation of flow cytometry data.

The flow lab is currently being managed by Lavanya Sayam.

Flow Cytometry allows the rapid acquisition of data from thousands to millions of particles. Each particle is measured for relative size, internal complexity and relative fluorescence intensity for up to 12 colors. We currently analyse samples from a wide range of sources from yeast and fungus, to the fly, rodents, pets and other larger animals. An example of some common assays we are currently running are shown below.

New Investigators:
We ask that all new investigator fill out a cytometry request form the first time they use the facility. Users with established protocols can just e-mail us with the run information. New users should estimate run time and prepare appropriate controls - see our sample prep page.

This core facility does not currently support unassisted use of the aria, but has a user operated LSRII. We can run and will sort the samples for you. There are other fee for use CU cytometry related instruments available on the CU campus.

The fees for use of the facility are:

All sorts on the FACSAria are assisted-
Cornell University (Vet College) - $60.00/hr plus a 1hr set-up charge for sorting runs.
Cornell University (non Vet.)$80.00/hr plus a 1hr set-up charge for sorting runs.

LSR II fees:

Cornell University (Stem Cell Group) - $40.00/hr
Cornell University (Vet.) Unassisted-$50.00/hr (available only to members trained by the Flow Lab)

Cornell University (Vet. and Stem Cell Group) assisted-$60.00/hr

Cornell University (non Vet.) Assisted-$80.00/hr

Cornell University (non Vet.) Unassisted-$60.00/hr (available only to members trained by the Flow Lab)

Non Cornell Assisted-$127.20/hr

All Runs Require	Negative unstained controls (0.5ml minimum volume)	Compensation controls for each fluor (0.5ml minimum volume)
Acquisition Samples	It is nice to have 0.5ml of sample volume with 1million cells per ml.	Data acquisition runs typically only acquire 10,000 to 30,000 gated events, but still require a minimum 250ul of sample volume.
Sort Runs	Cells should be at 20-30 million cells/ml (5,000 to 35,000 events/s)	The Aria can sort at up to 50- 100 million cells/hour (a flow rate of 0.2-0.8ul/s and up to 2ul/s with less resolution)
Sort Time	~1-2 ml of sample volume per hour. Collection rate(cells/hour) = 2(%Viability) X (%Positives) X (cell concentration)	Collection time = (Sort number target) X (collection rate) * not including set-up
Collection Tubes	Investigator should bring collection tubes	1ml, 5ml, or 15ml tubes- some cells require tubes to be coated with BSA/Serum or sorted into tubes containing media
Collection Tubes	1ml microtubes-Bio-Rad-223-9391 5ml (12X75mm)-BD Biosciences-352058 15ml conical-BD Biosciences-352196	Multiwell plates and glass slides 96,48,12,6 well plates max 1.5cm in height slides should be coated

Questions
Please direct all questions to Lavanya at (607)253-4004 lgs56@cornell.edu

Directions

The facility is located in the basement of the Vet. Research Tower down the far right corridor and around the left corner.

All samples with large amount of debris, dead cells or particulate material MUST
be filtered through nylon mesh of less than 70microns.

There is a charge of $2.00/sterile filter for samples filtered at the facilty or Cell filters can be purchased at:

Thomas Scientific	Sterile 40micron-50ml tube BD cell strainers	4620E99	$87.40/cs50
Thomas Scientific	Sterile 70micron-50ml tube BD cell strainers	4620F02	$87.40/cs50
Thomas Scientific	non-Sterile 70micron-6ml 12X75mm tubes	4620F40	$447.70/cs100
SEFAR America inc. 1-800-995-0531	NONsterile 41micron-sheets (Autoclave safe)	NITEX 03-41/31	$179.04/3 yards