BD FACSJazz

 

Single Cell Gene Analysis

Flow phenotype correlated with gene expression utilizing index sorting
Flow phenotype correlated with gene expression utilizing index sorting
Eighty-four single cells were selected at random to be sorted into a 96-well plate, with a BD FACSJazz, from an unequal mixed population of two cell lines, KG2A and Jurkat, while simultaneously capturing and recording their surface marker staining. Using the index sort feature of BD FACS Sortware, the surface phenotype of a single cell in any well coordinate can be referenced. Subsequent single cell analysis for gene expression (using the Fluidigm Biomark™ HD gene expression platform) for the 84 single cells demonstrated exact correlation between cell types predicted by both technologies. There are 6 single KG2A cells and 78 single Jurkat cells (well locations indicated). Antibodies used were CD2 APC and CD34 PE-Cy7.
Gene expression data (96 simultaneous gene expression targets for each cell) courtesy of Fluidigm Corporation, South San Francisco, CA, USA.

 

Stem Cell Sorting

Murine stem cell development progression
Murine stem cell development progression
BD FACSJazz™ analysis of murine stem cells expressing GFP with Flk1 reveals three distinct cell populations, GFP-Flk1-, GFP+Flk1-, and GFP+Flk1+cells, which represent a developmental progression ranging from pre-mesoderm to pre-hemangioblast mesoderm to the hemangioblast. GFP expression is a marker for mesoderm. The antibody used was Flk1 PE-Cy™7.
Data courtesy of Valerie Kouskoff, Stem Cell Haematopoiesis Group, Cancer Research UK, Paterson Institute for Cancer Research, The University of Manchester, England.
Adipose-derived stem cell GFP expression profiling
Adipose-derived stem cell GFP expression profiling
Adipose-derived stem cells expressing GFP were co-cultured with pancreatic tumor cells and run on the BD FACSJazz. The GFP-positive cells were sorted at 6,000 cells per second in 1.5-drop pure mode and collected into 15-mL tubes. Data courtesy of Dr. Karen Clise-Dwyer, Director South Campus Flow Cytometry & Cell Sorting Core Facility, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
Isolation of single hematopoietic stem cells from murine bone marrow
Isolation of single hematopoietic stem cells from murine bone marrow

Hematopoietic stem cells were identified in the bone marrow of a mouse expressing eGFP under the actin promotor. Sequential gating by scatter, then of singlets, lineage/c-kit+, Sca-1+, and CD150+CD48 events was used to deposit single GFP+ HSCs into 72-well Terasaki plates with the BD FACSJazz in 1.0 drop purity mode at a rate of 1,200 cells per second. The final plot shows the lineage/c-kit+/Sca-1+ cells back gated on the FSC vs SSC plot.

Antibodies used were CD150 PE, Lineage PerCP-Cy™5.5, Sca-1 PE-Cy™7, and c-Kit APC and CD48 APC-Cy7.


 

Neuroblastoma Cell Line Sorting

Human neuroblastoma cell line sorting
Human neuroblastoma cell line sorting
SK-N-BE 2 human neuroblastoma cells were analyzed for the expression of CXCR4 cell surface receptor using an APC-labeled antibody and run on a BD FACSJazz. Cells were sorted based on staining brightness (low, medium, and high) in 1.5 drop pure mode at a rate of 3,500 events per second. A purity check of the bright staining cells shows greater than 98% purity.
Data courtesy of Christopher Brownlee, Flow Cytometry Facilities Manager, Biological Resources Imaging Laboratory, The Mark Wainwright Analytical Centre, University of New South Wales, Sydney, Australia.
Samples courtesy of Dr. Jamie Fletcher and Mr. Ben Wilkinson, Childrens Cancer Institute Australia, Lowy Cancer Research Centre, UNSW, Sydney NSW 2052
Human neuroblastoma cell line sorting
Human neuroblastoma cell line sorting
SK-N-BE 2 human neuroblastoma cells were analyzed for the expression of CXCR4 cell surface receptor using an APC-labeled antibody and run on a BD FACSJazz. Cells were sorted based on staining brightness (low, medium, and high) in 1.5 drop pure mode at a rate of 3,500 events per second. A purity check of the bright staining cells shows greater than 98% purity.
Data courtesy of Christopher Brownlee, Flow Cytometry Facilities Manager, Biological Resources Imaging Laboratory, The Mark Wainwright Analytical Centre, University of New South Wales, Sydney, Australia.
Samples courtesy of Dr. Jamie Fletcher and Mr. Ben Wilkinson, Childrens Cancer Institute Australia, Lowy Cancer Research Centre, UNSW, Sydney NSW 2052

 

Advanced Immunophenotyping

Six-color immunophenotyping
Six-color immunophenotyping
Human peripheral blood lymphocytes stained with six cell-surface markers analyzed with BD FACS™ Sortware (BD FACSJazz with 488-nm, 640-nm, and 405-nm lasers).
A. Histogram
Histograms show cell counts for a single scatter or fluorochrome variable.
B. Dot plot
Dot plots show two variables plotted against each other, useful for identifying subpopulations.
C. Overlay histogram
Overlay histograms show cell counts for a variable, such as fluorescence, for multiple subpopulations.
D. Density plot
Density plots show two variables plotted against each other using color to illustrate heavily concentrated events.
E. Contour plot
Like density plots, contour plots show two variables plotted against each other, but instead use topographic lines to show the boundaries of populations.
F. Analysis of contour plot
Employ topographic lines to define subpopulations or move the cursor over a topographic line for instantaneous subpopulation statistics.



BioMark is a trademark of Fluidigm Corporation.

Cy™ is a trademark of Amersham Biosciences Corp. Cy™ dyes are subject to proprietary rights of Amersham Biosciences Corp and Carnegie Mellon University and are made and sold under license from Amersham Biosciences Corp only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from Amersham Biosciences Corp, 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.