Flow Cytometry Core

Resources

Salk Institute for Biological Studies - Flow Cytometry Core - Resources

Resources


The BD Biosciences FACSAriaTM Fusion is a high-end cell sorter. The Salk’s Aria Fusion is from the Special Order Research Product (SORP) line, with air launched high power lasers. The Aria design integrates fully fixed stream and optical alignment with a patented quartz cuvette flow cell and optical gel-coupled optics. While geared towards high sensitivity, complex flow cytometry, it remains easy to use, controlled by the same Diva software used by other BD Biosciences analyzers in the core. It is configured with 13 fluorescence and 2 light scatter detectors. This instrument is equipped with a full enclosure Class II Type A2 biosafety cabinet for sorting up to biosafety rating level BSL2.

The Aria Fusion has several nozzle sizes available (70, 85, 100, 130-um) to accommodate a variety of particle sizes and can sort up to 4 populations simultaneously. Cells may be collected into a variety of tube types or multi-well plates (e.g. 384 well).

CITING USAGE IN PUBLICATIONS: This instrument was acquired by a NIH Shared Instrumentation Grant (S10) award along with additional funding support from Larry and Carol Greenfield Technology Fund. In addition to the Salk’s Cancer Center grant NIH-NCI CCSG: P30 014195, please reference S10-OD023689 for reporting the use of this sorter.

The Salk core has 2 state-of-the-art BD InfluxTM cell sorters. These are jet-in-air design instruments uniquely suited for isolating delicate cells, and for high-speed sorting. “Influx-1” is equipped with 5 lasers, 11 fluorescent detectors. “ShadowFACS” is equipped with 4 lasers, 13 fluorescent detectors. Additionally, ShadowFACS is equipped with a forward scatter (FSC) pinhole assembly that facilitates detection of dim FSC signals when performing small particle analysis. Both Influxes have biosafety enclosures that enable BSL2 rated sorts. Several features of the Influx (including flow cell characteristics and use of low-noise pre-amp electronics) come together to enable excellent detection sensitivity and stable high-throughput sorting, while maintaining the integrity and viability of sensitive cell types. It is also remarkably fast and accurate at sorting into 384-well plates (~3.5min/plate for highly abundant targets).

The Influx has several nozzle sizes (70, 86, 100, 140-um) available to accommodate a variety of particle sizes and can sort up to 6 populations simultaneously. Cells may be collected into a variety of tube types or multi-well plates (e.g. 384 well).

The BD Biosciences LSR II is a configurable, high-end, analytical cytometer. A reliable platform well suited to complex polychromatic flow cytometry, the Salk’s Flow Core has two units equipped with 5 lasers, and the maximum 18 fluorescence detectors. Since the two are configured similarly, users are able to move easily from one instrument to the other. LSRII-2 is also equipped with a HTS (High Throughput Screening) module, which enables samples to be acquired in multi-well plates. The LSR II cytometer runs on BD FACS Diva acquisition software. Both units are located in the lower NE basement: LSRII-1 is located in the STEM Core, while LSRII-2 located across the corridor in the main FCCF Sorter Room.

The Becton-Dickinson FACSCanto II™ cytometer is an easy to operate 3-laser benchtop analyzer capable of detection of up to 8 fluorescent parameters and 2 light scatter parameters. The sample injection tube (SIT) features an automated flush function that rinses the probe between tubes to reduce sample cross contamination. The FACSCanto II™ runs on BD FACS Diva acquisition software. It is also equipped with a fluidics cart that holds all fluids necessary to operate, clean, maintain the instrument, as well as corresponding automated commands within the FACS Diva software to assist users in automatically starting up and shutting down the instrument in proper running buffer or fluidics shutdown fluids.

The MACSQuant VYB is a user-friendly compact bench-top analyzer with 3 lasers and 8 fluorescence detectors. The VYB has several convenient automated features including start up, cleaning and priming, flushing between samples, and a complete walk-away shut down protocol that can be set to self-execute at a specific time after start up. Buffer tanks (with fluid sensors that have software integrated warning LEDS) are attached to the instrument sides for easy access.   The VYB uses a robotic syringe needle for sample uptake, which allows delivery of a defined sample volume with high precision. This type of volume-defined sampling allows absolute cell counts (error margin ±5%) to be directly determined during acquisition. The Salk core’s instrument is also equipped with a motorized “MiniSampler” module, allowing for easy transition between single tube analysis and automated acquisition. After establishing settings, users can run a rack of sample tubes or a micro-titre plate (e.g. 96-well). Racks can be refrigerated ahead of use; these are designed to maintain 4C degrees for several hours, until a phase change back to liquid occurs (Chill 5 for 5ml tubes, and Chill 96 for plates).