Analytical Flow Cytometry
Flow Cytometry is an important tool in all aspects of cell biology research. Many different combinations of antibody-based stains, fluorescent protein expression, and viability indicators are measured at the single cell level. The Core provides access to cytometry equipment and staff with expertise to assist researchers in developing assays. Basic analytical flow cytometry instrumentation at the Core includes the BD FACSCanto II, BD FACSCelesta, Beckman Coulter Gallios, and BD FACSCaliber.
High Parameter Analytical Flow Cytometry
Flow cytometry instrumentation in the Core can detect up to 35+ fluorescent parameters at once on the single cell level. The Core staff assists researchers in developing high parameter assays, including panel design, instrument set up, assay troubleshooting, data acquisition, and analysis. Users with complex assays are encouraged to contact Core personnel during the planning stages of their experiments. High parameter instrumentation at the Core includes the BD LRSFortessa, two BD LSRFortessa X-20s and Cytek Aurora.
We use electrostatic droplet based high-speed cell sorting to isolate a wide variety of cells based on many different combinations of fluorescence markers. Several nozzle size/pressure configurations are available. Up to 16 color cell sorting is provided with a variety of options for collection vessels including various test tube sizes and multiwell plates. Cell sorting can entail isolation of single cells as well as tube-based collection of up to 6 simultaneous populations for RNASeq, culture, or many other downstream applications.
Imaging Flow Cytometry
The Amnis ImageStreamX Mark II is an imaging flow cytometer that uses CCD cameras to image cells in flow with up to 9 fluorescent channels for each cell. A number of assays have been developed for the ImageStreamX, including colocalization, nuclear translocation, apoptosis assays, calcium flux assay, characterization of diffuse vs. punctate staining, assays of immune cell interactions, extracellular vesicle detection, circulating tumor cell detection, cell counts and transfection efficiency.
High Parameter Data Analysis
Analysis of multiparametric cytometry data can be time-consuming and complex. The Core provides expertise and access to software with modeling and computational capabilities to analyze high parameter, single cell data sets. Users with 12+ parameter data and imaging cytometry data are encouraged to consult Core staff for data analysis. The Core has software available for high dimensional analysis, including FlowJo, FCS Express, and Cytobank. Core software for imaging cytometry analysis includes IDEAS and FCS Express.
Custom mAb Conjugations
The Core performs conjugations primarily for monoclonal antibodies that are not commercially available as conjugates. The core can prepare 100 microgram-scale antibody conjugations with cell-free QC. Available fluorochromes include AlexaFluor Dyes (488, 647, 555, 594, etc.) and Pacific Blue. Contact Kathryn Cox at firstname.lastname@example.org for information.
Rates for MD Anderson Cancer Center Faculty
|Service||Instruments||Assisted Rate||Unassisted Rate||Unit|
|Analytical Flow Cytometry
||Canto II, Celesta, Gallios, Caliber||$70||$50||Hour|
|High Parameter Analytical Flow Cytometry||Fortessa, Fortessa X-20||$75||$65||Hour|
|Cell Sorting||Aria III, Aria Fusion, Influx, Jazz||$85||$60||Hour|
|Imaging Flow Cytometry
|High Parameter Data Analysis||N/A||$22||N/A||Hour|
|mAb Conjugations||N/A||$130||N/A||0.1mg Reaction|