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Cell Sorting

FCC facility staff will perform all sorting on the BD FACSARIA3 and BD FACSFusion . All new users are required to have an initial consultation with facility staff prior to use of the cell sorter, and all users are required to fill out a sort request form prior to each experiment. If necessary, biocontainment and biosafety concerns must be addressed by the Scripps Florida Biosafety Office.

A discussion on sample preparation, appropriate buffer conditions, collection tubes, instrument controls, experimental controls, etc., will be part of the initial consult required for first time users.

Cell Sorting Guide

Sorting on the BD FACSAria3

The BD FACSAria3 is a four-laser system configuration. The FACSAria3 uses a 488nm (blue), 561nm (yellow-green), 633nm (red), and a 407nm (violet) laser system. Sorting is available using up to sixteen fluorescence parameters as well as light scatter discrimination (cell size, organelle composition and density, and doublet discrimination). Applications include high speed, four-way cell sorting, cell purification, slide sorting, and cell cloning (by automated deposition into 6, 12, 24, 48, 96 well plates).  Default and optional configurations for different fluorescence parameters and combinations can be viewed here.

ARIA Instrument Configuration

Sorting on the BD FACSFusion

The BD FACSAria Fusion is a four-laser system configuration. The FACSAria Fusion uses a 488nm (blue), 561nm (yellow-green), 640nm (red), and a 405nm (violet) laser system. Sorting is available using up to sixteen fluorescence parameters as well as light scatter discrimination (cell size, organelle composition and density, and doublet discrimination). Applications include high speed, four-way cell sorting, cell purification, slide sorting, and cell cloning (by automated deposition into 6, 12, 24, 48, 96 well plates). The FACSAria Fusion is enclosed in a ClassII Type A2 Baker biosafety cabinet consistent with BSL2 containment.

Fusion Instrument Configuration