Equipment

Instruments available for cell sorting:

BD FACSAria Fusion sorter

The BD Biosciences FACSAria Fusion™ is a high-end Special Order Research Product (SORP) cell sorter configured with 4 high power lasers (Violet 405 nm, Blue 488 nm, Yellow-Green 561 nm, Red 637 nm), 17 fluorescent detectors and 2 light scatter detectors.

The BD Biosciences FACSAria Fusion is capable of sorting up to 4 populations simultaneously. Samples can be sorted into 12 x 75 mm FACS tubes or 1.5 ml Eppendorf tubes for 4-way sorts, and into 15 ml centrifugation tubes for 2-way sorts.

In addition, the FACSAria can deposit single cells into 96- or 384-well plates. The optional index sorting function ensures that a single-sorted cell has the desired phenotype. This system maintains sample sterility and can sort up to ~25,000 events per second depending on cell type and nozzle size (70, 85, 100, and 130 um). 

The Fusion sorter is housed in a laminar flood hood for BSL2 work (noninfectious, human cells).

Instruments available for flow cytometry:

BD FACSymphony A3 Flow Cytometer

The BD FACSymphony™A3 Cell Analyzer is a novel high parameter system that offers qualitative and quantitative analysis of cells and other particles using multi-parametric analysis. This instrument is configured for five laser (UV 355 nm, Violet 405 nm, Blue 488 nm, Yellow-Green 561 nm, Red 637 nm) analysis and can simultaneously acquire and distinguish between 28 different fluorescent signals in addition to the forward and side scatter parameters allowing for a 20+parameter definition of the cell.

The FACSymphony is operated using FACSDiva 9.1v software. FCS file versions 2.0, 3.0, and 3.1 can be exported for data analysis.

BD FACSymphony A5SE Spectral flow cytometer

The new BD FACSymphony™ A5 SE Cell Analyzer is a fluorescence-activated, spectral-enabled cell analyzer that offers researchers the ability to choose between spectral or compensation-based cell analysis to meet different flow cytometry needs. The BD Symphony A5 SE offers our highest level of simultaneous parameter detection by utilizing five lasers (including UV) and 48 detectors for maximum coverage of the fluorochrome emission spectrum.

The software used by this instrument is comparable to that used by the Symphony A3 and Aria Fusion, with the additional ability for spectral unmixing.

Amnis Imagestream Imaging Flow Cytometer

Amnis Imagestream-X MarkII imaging flow cytometer provides users with the ability to gain detailed images of a large number of cells in a relatively short period of time and with the opportunity to perform a range of novel applications including co-localization, internalization, stem cell differentiation, and cell-cell interactions.

ImageStreamX MarkII produces up to 12 high resolution images of each cell directly in flow, at rates exceeding 1,000 cells per second, and with the fluorescence sensitivity of conventional flow cytometers. These capabilities allow you to quantitate cellular morphology and the intensity and location of fluorescent probes on, in, or between cells, even in rare sub-populations and highly heterogeneous samples.

The Imagestream includes 3 lasers (405 nm, 488 nm and 640 nm) and can visualize up to 12 channels for unmixing fluorophores, and enables visualization of cells using 20X, 40X or 60X magnification.

Microscope for imaging:

Nikon A1 Confocal

The Nikon A1 laser scanning confocal microscopy enables diffraction-limited imaging of samples. Using 4 lasers (405, 488, 561 and 640 nm), and one of two detectors:

• Conventional 4-channel detector with 2 high dynamic range PMT's, and 2 GaAsP dectors

• 32-channel spectral detector, to allow spectral unmixing of similar fluorophores (or > 4 channel imaging)

Live cell imaging: The Nikon confocal is housed on a Ti-E stand with Perfect Focus System, and includes a live cell chamber for provision of humidified 5% CO2 (in air) on a heated insert, for long term (hours to days) imaging. Live cell imaging allows users to perform FRAP, FRET, FLIP and other similar experiments.