SDSC meeting sponsored by DVS (a Fluidigm Company) 4/09/14
Carina Torres and Cheryl Kim organized a great group of speakers for this month’s flow cytometry meeting. After introductions, we were reminded of Cyto 2014, which occurs next month in Ft. Lauderdale. There will be many great talks and courses over this week long conference. Carina also asked the group if we would be interested in attending a So Cal meeting, if held in San Diego, since the meeting normally takes place in the LA area. There was a very positive response.
The speakers for this meeting were Theresa Russel from Fluidigm and Anita Kant from DVS. The final speaker was Dr. Greg Behbehani who uses a mass spec cytometer in his research within Garry Nolan’s lab, where they worked closely with DVS to develop this technology. DVS, known for the revolutionary CyTOF mass spec cytometer, recently joined forces with Fluidigm to create a synergy of technologies for multidimensional single cell analysis.
Theresa Russel Every Cell is Unique –Single Cell Genomics
In traditional genomic studies, there is only a broad overview of the whole population. Up until now, the way scientists were able to obtain single cell genomic data was to sort individual cells using expensive and difficult to use cell sorters. These single cells could be sorted into 96-well plates for genetic analysis. Using a microfluidic system, such as the Fluidigm C1, scientists can isolate single cells in a 96 “well” format, isolate DNA and used it for “whole exome, whole genome, or targeted sequencing.” The C1 has a clear microchip format allowing visual as well as quantitative verification of single cell isolation.
Anita Kant Understanding Biological Heterogeneity through Mass Cytometry
How the CyTOF works:
An antibody or nucleic acid binder is conjugated to heavy metals of a particular atomic mass. These antibodies are bound to cells and ionized in the instrument. The metals are quantitated, relating to which antibody and how much was bound to the cell. DVS has over 250+ preconjugated antibodies. Labeling kits are available for scientists to label their own antibodies. Metals in the atomic mass range of 156-176 are ideal for this application.
Using high dimensional panels of up to 30 colors, the CyTOF2 can discriminate multiple pathways in a cell. CytoBank’s SPADE (spanning tree progression analysis of density-normalized events) software allows for a deep understanding of how genes are related within a disease and can be shown visually. Patients can be screened for cell signatures and monitored for changes with drug treatment.
Using mass spec cytometry to perform single cell systems biology analysis of MDS and AML.
Dr. Behbehani used mass spec cytometry to study biomarker changes in patients. Using the SPADE software, he was able to study MDS and AML patients’ blood for loss of hematagones and unknown biomarker expression. He used a barcoding system to help eliminate any doublets and analyzed the clusters found among the patients. Cell cycle studies using the CyTOF are made possible using iododeoxyuridine. Combining the DNA intercalator and other markers, such as the retinoblastoma protein and cyclins, he was able to study chemotherapy responses in different cell types and help guide individual patient drug therapies.
This is a great venue to discuss emerging technologies and research conducted in the flow cytometry community. Thanks to Carina and Cheryl for organizing this event.