For information about the MPRI Flow Cytometry Core or FACSAria, contact:
Ken Class, Director, Imaging and Flow Cytometry Facility email@example.com
2102 Biosciences Research Building College Park, MD 20742
Office: 301.405.4593 Lab: 301.405.0398
General laboratory operating hours: 10-6 Mon-Fri (extended hours available on request).
All signups are implemented using Google Calendar. You will receive an invite to the FACSCanto Google calendar once a valid gmail account is provided. You must continue to confirm dates/times with the facility director until 24/7 access to the facility has been granted. All FACSAria experiments must be arranged through the facility director.
You will be charged for time signed up for unless canceled 24hrs in advance.
|Instrument||Univ. Of MD.||External||**Min Billable Time|
|FACSCanto II||$20/hr||$50/hr||0.25 hr|
|FACSAria II||$40/hr||$50/hr||1.00 hr|
|FACSAria II sort setup||add 0.50 hr|
** Minimum Charge of $50 for External users on FACSaria and FACSCanto
All chargebacks are calculated on a 0.25hr basis with the exception of the Luminex instrument. Samples will be run and data collected by the facility Director, Ken Class. Frequent regular users who wish to have 24/7 access to the FACSCanto will be able to do so after undergoing one-on-one training and demonstrating proficiency in instrument operation.
All samples to be run on the FACSAria must be pre-filtered through no greater than 70um strainers prior to being run on the instrument. (BD Falcon tubes, cat# 352235 are a preferred source. The flow lab has samples of these to try, but it will be mandatory to eventually purchase your own).
Up to 9-color measurement of surface and/or internal antigens on FACSAria (8-color on FACSCanto)
(Cell Sorting) into tubes or tissue culture plates.
Data analysis programs available: FACSDiva software, Flowjo (coming soon), Microsoft Office products.
Excitation sources available and commonly used fluorochromes:
|Coherent® Sapphire™ solid state||488nm (blue)||FITC, GFP, Alexa Fluor-488, PE, PE-Texas-Red, DSRedPerCP, PerCP-Cy5.5, PE-Cy7, PI, 7AAD|
|JDS Uniphase™HeNe air cooled||633nm (red)||APC, APC-Cy7, Cy5, Alexa Fluor 647, Topro-3|
|Point Source Violet solid state||407nm (violet)||
Alexa Fluor- 405 and
-430, Cascade Blue, Pacific Blue, Dye Cycle Violet
|Contact Ken at firstname.lastname@example.org regarding any additional dyes that are not mentioned in the table above.|
|Nozzle tip size||Approx. max Flow Rate and operating pressure (cells/second)||Sample Concentration (cells/ml)*|
|70mm (70 PSI)||22,000||20-40x106|
|85mm (35 PSI)||15,000||10-20x106|
|100mm (20 PSI)||7,500||5-10x106|
|130mm (10 PSI)||3,000||1-5x106|
** For sorting experiments, it is preferable to prep cells toward the higher end of the concentration range if possible. Extra media can be brought to the sorting facility for diluting the sample if necessary.
** For “analysis only” experiments, 0.5-5.0x106cells/ml are the optimal concentration regardless of nozzle tip size.