Flow Cytometry Facility

Overview

The Flow Cytometry Shared Resource provides investigators with the technological resources and professional assistance for high quality, multiparameter flow cytometry analyses and sorting. The Facility is capable of cell sorting (sterile, at speeds up to 30,000 cells/sec) from homogeneous or mixed cell populations based on up to 32 fluorochromes, sorting up to six separate populations simultaneously, including human-derived samples at BSL-2 level. Facility personnel aid investigators in creating efficient and cost-effective experimental designs, through optimizing cytometry-specific reagent and fluorochrome selection, and offer assistance in operation of analysis instruments. Technical support is also provided for analyses of flow and imaging cytometry data for publication, presentation, and inclusion in grant applications, management of cytometric data (storage, archiving, and retrieval), and management of a site license for low-cost post-acquisition analysis software.

Services

• Multi-parameter flow cytometry analysis (up to 28 color) supporting experimental protocols.
• Fluorescence cell sorting (sterile) at rates up to 30,000 cells/second, sorting up to six populations simultaneously, utilizing 16 (FACSAria), 32 (MoFlo Astrios), or 9 fluorescent detectors (FACSMelody), all under BSL-2 conditions if necessary. The MoFlo Astrios has the capacity for UV-excited fluorochromes, including those for side population stem cell analysis and sorting. Both the MoFlo Astrios and FACSMelody can perform clonal sorting into plates.
• Imaging Cytometry – Amnis ImageStream^X analyses: Assistance provided for instrument operation as well as in developing appropriate staining protocols/techniques for assays including molecule co-localization, nuclear translocation, and localization of target molecules to intracellular organelles; and performing acquisition and analysis of experiments.
• Training and Consulting: Training of investigators on “user-friendly” flow cytometry instrumentation. Consulting, assisting in development of appropriate staining protocols/techniques.

Equipment & Features

Master Configuration/Fluorochrome List

Analysis Instruments:

• FACSymphony analyzer with high-throughput sampler – 28 color, 5 lasers
• LSRII analyzer – 18 color, 4 lasers
• LSRII analyzer – 14 color, 4 lasers
• FACSCelesta analyzer with high-throughput sampler – 12 color, 3 lasers
• FACSCalibur analyzer – 4 color, 2 lasers
• Amnis ImageStreamx imaging flow cytometer

Cell Sorting Instruments (available at BSL2+ level):

• FACSAriaII high-speed cell sorter – 16 color, 4 lasers
• MoFlo Astrios high-speed cell sorter – 32 color, 7 lasers
• FACSMelody user-friendly cell sorter – 9 color, 3 lasers

Pricing

For pricing information, visit iLab or contact the managing director.

This facility is supported in part by a Cancer Center Support Grant (CCSG) awarded by the National Cancer Institute (NCI) to The Wistar Institute Cancer Center.