NJMS Flow Cytometry and Immunology
||Sukhwinder Singh, PhD
||Patricia Fitzgerald-Bocarsly, PhD
Sukhwinder Singh: firstname.lastname@example.org,
The Flow Cytometry Core Lab provides support to the NJMS research community as well as to surrounding universities and private companies for applications involving flow cytometric analysis, cell sorting and imaging flow cytometry.
Flow cytometric analysis:
- BD FACSymphony A5 SE: The Symphony is equipped with 5 lasers (355nm, 405 nm, 488nm, 561nm, and 637nm). It is set up to acquire 48 fluorescent parameters either conventionally or in a spectral format, which will allow for the acquisition of virtually any fluorophore able to be excited by its five lasers. The Symphony is ideal for producing high-dimensional data sets of panels consisting of 20+, 30+ and even 40+ markers. In the conventional setting, there are 10 parameters off the 355nm, 14 parameters off the 405nm, 10 parameters off the 488nm, 9 parameters off the 561nm and 6 parameters off the 637nm laser.
- BD LSRFortessa X-20: The Fortessa is equipped with 5 lasers (355nm, 405nm, 488nm, 561nm, and 642nm). It can acquire up to 18 fluorescent parameters per sample with 3 colors off the 355nm, 5 colors off the 405nm, 2 colors off the 488nm, 5 colors off the 561nm and 3 colors off the 642nm laser. Since the instrument is equipped with the 561nm laser it has the ability to excite RFP, TdTomato and mCherry, as well as other fluorescent proteins. The Fortessa is also equipped with a 96-well plate high throughout system for running plates.
- ThermoFisher Scientific Attune NxT: The Attune is equipped with 4 lasers (405nm, 488nm, 561nm and the 637nm). It can acquire up to 14 fluorescent parameters per sample with 5 colors off the 405nm, 2 colors off the 488nm, 4 colors off the 561nm and 3 colors off the 637nm laser. The Attune is also equipped with a 96-well pate system for running plates. (Located in the NJMS Cancer Center)
- BD LSR II: The LSRII is an air-cooled four-laser benchtop flow cytometer with the ability to acquire 10 colors. It includes a solid-state 488nm (blue) laser, a JDS Uniphase 633 HeNe laser (red) and a Coherent 405nm laser (violet) and a solid-state 355nm laser (UV) with the ability to detect 5 colors from the blue laser, 3 colors from the red laser and 2 colors from the Violet laser utilizing fiber optic technology.
- BD Accuri C6: The Accuri is equipped with two lasers and is capable of acquiring 4 fluorescent parameters (3 off the 488 and 1 off the 640nm). (Located in the NJMS Cancer Center)
- BD FACSAria II SORP and BD FACSAria Fusion: Both Sorters are equipped with four lasers (355nm (UV), 488nm, 561nm and 640nm), allowing for 18 marker-based sorting for clonal expansion, multi-well plating, sorting onto slides and simultaneous 2, 3, or 4 population sorting. With the addition of the 561 nm laser, the sorter is capable of sorting cells expressing the red fluorescent family of proteins including RFP, mCherry, mPlum and others. It is housed in a Baker biosafety cabinet and in addition has an aerosol containment unit built-in, which allows for the sorting of BSL2 agents and virus-infected cells.
- BD FACSymphony S6 Cell Sorter (BSL-3): The Symphony S6 is located in the International Center for Public Health (ICPH) Regional Biocontainment Laboratory (RBL) and operated as a satellite of the FCICL. It allows for the analysis and sorting of live TB and other agents that require BSL-3 handling along with the ability to analyze the interaction between mammalian cells and those pathogens. The Symphony S6 is equipped with four lasers (405, 488, 561 and 641 nm) allowing for the simultaneous acquisition of up to 38 parameters. The Symphony S6 is capable of sorting 6 populations simultaneously or deposit the cells into multi-well plates, as well as the ability to sort cells onto slides.
Imaging Flow Cytometry:
- AMNIS ImagestreamX MarkII: The FCICL operates an Amnis ImageStreamX MarkII (Amnis Corporation) imaging flow cytometer that is equipped with four lasers (405, 488, 561, and 642 nm). This instrument captures high-resolution digital images of cells in flow at rates of approximately 1,000 – 2,000 cells/per second. It simultaneously acquires up to 12 different images of each cell, which includes 10 colors of sensitive fluorescence imagery, side scatter and brightfield imagery. The ImageStream combines the quantification and analysis of cellular morphology with the ability to perform all the fluorescence intensity measurements of conventional flow cytometry, allowing for rapid analysis and classification of thousands of cells using the ImageStream IDEAS software. The IDEAS software calculates over 100 parameters per cell, including all the standard intensity-based parameters and statistics employed in flow cytometry as well as numerous morphological parameters such as cell area, perimeter, aspect ratio, texture, spot counts and internalization ratios. The ImageStream is ideally suited for quantitative investigations of nuclear translocation, phagocytosis, intracellular co-localization, apoptosis, cell: cell interactions and fluorescent in situ hybridization in suspension.
- Luminex xMAP INTELLIFLEX: The Intelliflex is a new flow-based multiplexing platform that is based on the proven xMAP technology from Luminex. The system is capable of analyzing 500 analytes in a single sample volume. It is equipped with a 532 nm laser with a variable power output based on the mode selected with a maximum power output of 50mW with a dynamic range of 5.5 logs. The system is capable of running a 96-well plate (20 min runtime) or a 384-well plate (75 min runtime). Numerous kits are available from multiple vendors to analyze cytokines and growth factors to phosphorylated proteins. Contact Dr. Singh for Vender representative contacts.
- Beckman Coulter AcT diff. The AcTdiff is a hematology instrument that can provide a 3-part differential. It only requires 18 uL of whole blood for analysis.
Software resources :
- Additional software available for use in multiparameter studies includes BD Bioscience FACSDiva for data analysis on our analyzers. Modfit LT software for DNA and cell proliferation analysis. For the ImageStream, the IDEAS software package is available which can be used in the Flow Cytometry Core Lab at the additional ImageStream workstation. The IDEAS software is also available to NJMS researchers for use in their laboratories through our site license.
- The Core Laboratory also administers a university site license for FlowJo analysis software. Please go to the Charges and Policies link below for pricing and availability.
Equipment time and services at the Flow Cytometry Core Lab are available to investigators on an hourly fee-based schedule. Please click the link below for fees.
Charges and Policies