Flow Cytometry is done on a fluorescence activated cell sorter (FACS) Aria III instrument (BD Biosciences). The Aria III is equipped with blue (488 nm), green (561 nm), red (633 nm) and violet (405 nm) lasers with 11 detectors that can measure up to 10 colors simultaneously. We are currently unable to run un-fixed human or primate cells as the Aria III instrument is not in a BSL2+ facility. Formalin fixed human or primate cells may be analyzed.
Flow cytometry has many uses, including cell cycle, cell viability, redox state, and immunophenotyping to name a few. Core personnel will assist the customer with strategy and provide guidance to minimize the need for compensation and to improve your analysis. We will help with data interpretation as well. The Aria III utilizes FACSDiva software and we also have FloJo which is useful for cell cycle analysis.
The Aria III is calibrated each day of use with CS&T calibration beads (BD Biosciences) to ensure voltages and %rCVs are within acceptable ranges (e.g. bright bead %rCV <6%). For determining significance of a biological response, multiple controls and experimental samples must be assessed using standard statistical methods. Isotype controls, fluorescence minus-one (FMO) controls, and compensation controls should be used as deemed necessary by the Core facilitator. Rare events (<5%) are collected according to Poisson statistics to a desired %rCV using the equation CV=100/SQRT (#rare events).
Standard Operating Procedures
Have Questions? Contact Us.
|Qing Shi, Lab Manager
Metabolic Molecular Phenotyping Core-Chapel Hill Facilities
T: (919) 966-7162
|Steve Orena, Lab Manager
Metabolic Molecular Phenotyping Core-Kannapolis Facilities
T: (704) 250-5041