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> Flow Cytometry Cell Sorting
UMDNJ-NJMS Flow Cytometry Core Laboratory
| Technical Director: |
Dana Stein, BS, MT, ASCP |
| Scientific Director: |
Patricia Fitzgerald-Bocarsly, PhD |
| Contact info: |
Dana Stein: steinda@umdnj.edu,
973- 972-7502
Patricia Fitzgerald-Bocarsly:
bocarsly@umdnj.edu, 973-972-5233
|
| Location: |
MSB F522 |
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.
Resources available
Flow cytometric analysis:
Three BD Biosciences FACSCaliburs
each having a dual laser capacity (an air-cooled 488 nm argon laser and a red
diode laser) with capabilities to do up to four color analysis and an HTS sampler
for multiwell sampling. One of the BD FACSCaliburs is located in the NJMS �
University Hospital Cancer Center.
LSR II air-cooled four-laser
benchtop flow cytometer (BD Biosciences Immunocytometry Systems) with the ability
to acquire 12 colors. BD digital software (DIVA) provides both online and offline
compensation running in a Windows based environment. It includes a solid-state
488nm (blue) laser, a JDS Uniphase 633 HeNe laser (red), a Coherent 405nm laser
(violet) and a sold-state 355nm laser (UV) with the ability to detect 5 colors
from the blue laser, 3 colors from the red laser, 2 colors from the Violet laser
and 2 colors off the UV laser utilizing fiber optic technology.
Cell sorting:
FACSVantage SE TM with the
FACSDiVa upgrade cell sorter, which provides high performance, high speed, multi-color
(up to 7 markers) cell sorting, with capability for single cell sorting into
multi-well trays for clonal expansion. This cell sorter from BD Biosciences
has a water-cooled Enterprise Laser that has a 488nm argon and UV component
and an air-cooled HeNe laser for use when more complex studies are required.
A Cytek Aerosol Containment System is also available for the cell sorter.
Imaging Flow Cytometry:
Amnis ImageStream (Amnis
Corporation) imaging flow cytometer that is equipped with 488 nm and 658 nm
lasers and an extended depth of field module. This instrument, which operates
in a Windows-based environment, introduces a new technology platform that captures
high-resolution digital images of cells in flow at rates of approximately 100
cells/per second. It simultaneously acquires up to six different images of each
cell, including four 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.
Software resources :
Additional software available
for use in multiparameter studies includes BD Bioscience CellQuest for the Caliburs,
Modfit LT software for DNA and cell proliferation analysis, HTS software for
the HTS sampler, and BDDiva software for the LSRII and the Cell sorter. 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 UMDNJ-NJMS researchers for use in their laboratories
through our site license.
Facilities use
Equipment time and services at the Flow
Cytometry Core Lab are available to investigators on an hourly fee-based schedule.
Please contact the Flow Cytometry Core Lab for more information.
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