Thursday, 29 November 2018

New cell sorting possibilities at CIBIO Cell Analysis and Separation Core Facility

The detection of Brilliant Violet dyes and the optimal resolution of many red fluorescent proteins are now available

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The detection of Brilliant Violet dyes and the optimal resolution of many red fluorescent proteins are now available.
Cell sorter has been recently upgraded from BD FACSAria II to BD FACSAria III and equipped with two new lasers, violet (405 nm) and yellow green (YG, 561 nm) ones.

This upgrade will enable to increase the applications of the existing instrument, maintaining the good and well-known proprieties of Aria’s family cell sorters. Now, the instrument has 5 lasers (375nm/405nm, 488 nm, 561 nm, 632 nm) and measures a maximum of 14 colors simultaneously. For optical configuration and examples of fluorochromes see downloaded attachment.
At the heart of the BD FACSAria III is a new quartz cuvette flow cell in true fixed alignment with the laser and gel coupled to the collection optics. The benefits are many from a greater sensitivity in multicolor approach to an improved resolution for side population applications and DNA cell cycle analyses. The violet laser gives more color choices for multicolor panel design as well as an expanded number of bright dye available, such as Brilliant Violet reagents. These new dyes are characterized by enhanced brightness to resolve at the best the dim signals.
The laser YG is a great advantage to excite and detect PE-conjugated antibodies and fruit fluorescent protein such as mCherry. This laser increases PE detector sensitivity and minimizes or eliminates FITC spectral overlap into the PE channel, giving the instrument the best chance at resolving a dim staining PE population.
Furthemore, the software was upgraded to a new version of BD FACSDiva that includes the possibility to do index sorting. Index sorting allows to sort single cells onto a plate or slide with retrospective identification of each single cell's collected morphology and fluorescence parameters. The new sorting possibilities move us to high parametric cell analyses and sorting and allow single cell index sorting for a better understanding at the single cell level.

The instrument’s upgrade was funded by European Regional Development Fund (ERDF) 2014-2020.