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RNA Flow Cytometry



Presented by

Steven McClellan

Flow Cytometry Core Laboratory, Mitchell Cancer Institute, University of South Alabama, Mobile, AL

&

Paul K. Wallace

Department of Flow & Image Cytometry, Roswell Park Cancer Institute, Buffalo, NY



August 26, 2015

9:00 am PT US/Canada
12:00 pm ET US/Canada
5:00 pm GMT/UK/Portugal

Register: http://cytou.peachnewmedia.com
Cost: FREE

Steve McClellan, BS, MT(ASCP) is a life science professional with over 25 years of expertise in advanced flow cytometry & cell sorting, as well as basic and clinical research in the areas of cancer biology, stem cell therapy, transplant immunology and xenotransplantation. He is currently Manager of Basic & Translational Research Operations at the University of South Alabama Mitchell Cancer Institute, where he also serves as Chief, Flow Cytometry & Imaging Core Laboratories. For the past seven years, his lab has been conducting research on cancer stem cells; working to develop better methods of purification, culture and analysis at both genetic and functional levels, as well as using cancer stem cells in HTS drug discovery.

Paul K. Wallace, PhD has served since 2003 as Director of the Flow and Image Cytometry Department and is Professor of Oncology at Roswell Park Cancer Institute (RPCI) in Buffalo, NY. Under his direction, the Flow and Image Cytometry Department at RPCI offers a strong combination of clinical and research missions.  The department's clinical emphasis is on the diagnosis and monitoring of patients with leukemia and lymphoma. He is also Associate Professor of Microbiology & Immunology, Dartmouth College, Hanover, NH and Associate Professor of Biotechnical and Clinical Laboratory Sciences, University at Buffalo, Buffalo, NY. He is President elect of the International Society for Advancement of Cytometry and a Councilor of the International Clinical Cytometry Society.
Webinar Summary:
Flow cytometry permits the simultaneous measurements of many biomarkers in individual cells from bulk populations. Until now analysis has been limited, however to primarily analysis of proteins and total DNA or highly abundant DNA sequences. Since most RNA gene transcripts are present at very low quantities our ability to detect these mRNA species by flow cytometry has been limited. In 1993, Patterson et. al. (Science, 1993. 260:976) used a PCR driven in situ hybridization technique to detect HIV mRNA in infected cells, however the stringent conditions required by this technique prevented its wide spread application to and development for flow cytometry. Two new techniques, PrimeFlowTM (Affymetrix/eBioscience) and SmartFlareTM (EMD Millipore) have recently been commercially introduced and are seeing increasing use among research scientists.

The PrimeFlow RNA assay system allows for the determination of differential RNA expression within a mixed population of cells. The assay improves sensitivity and lowers background of fluorescent in situ hybridization (FISH) through the use of a branched DNA signal amplification. In contrast, the SmartFlareTM system employs a gold nanoparticle that is actively endocytosed into most cell types. In the cytoplasm, interaction with the specific mRNA target causes the generation of a fluorescent signal, which can be detected by flow cytometry or microscopy. Protocols and data will be presented showing the usefulness of both systems. By the end of the webinar, participants should have an understanding of which technique is most appropriate for their applications; how to apply them to their research and most importantly gain an understanding of the pitfalls and solutions from individuals who have learned these the hard way.

CYTO U Webinar Recordings
A recording of this webinar will be posted online at CYTO U within 24-48 hours after the live event for free viewing by all.  You may also want to watch the recordings of CYTO U's last three webinars, Predicting the Best Resolution and Sensitivity in Panel Development and Reducing Inter-instrument Variability in Flow Cytometry by Steve Perfetto and Jim Wood, Business Continuity and Risk Mitigation for Shared Resource Core Laboratories by Sheenah Mische and Kinetic Image Cytometry of Stem Cell-Derived Cardiomyocytes and Neurons by Jeffrey Price at CYTO U.  For more details, visit: http://cytou.peachnewmedia.com.

CYTO University
CYTO University is an online educational resource created by ISAC for its members and the wider cytometry community. In addition to free webinars, CYTO U presents recorded courses and tutorials from the CYTO Conference, and interactive online courses on a variety of cytometry topics. These are available at no cost to ISAC members and for a nominal charge for non-members. Webinars are free for all. Learn more at http://cytou.peachnewmedia.com

CYTO U Access for ISAC Members
For current ISAC members, your user login and password to CYTO U are the same user login and password you have for the members-only section of the ISAC Web site. If you remember your user log in to the ISAC Web site but cannot remember your password, you can obtain your password from the ISAC Web site by clicking on the link "forgot your password?".  If you cannot remember your username, you can contact [log in to unmask]<mailto:[log in to unmask]> for your login information.


____________________
Kanika F. Pulliam, Ph.D.
Education Manager
International Society for Advancement of Cytometry (ISAC)
9650 Rockville Pike
Bethesda, MD 20814
(301) 634-7457
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