April 9, 2020
11 am to 12 pm EDT
Sponsored by
Webinar Description:
Three of the most respected researchers in their field will discuss the methods they are using to isolate single cells from suspensions, tissue, and from living cultures. Everyone whose work will be advanced
by using the most modern methods of cell isolation will benefit by attendance at this fast-paced set of talks.
Learning Objectives:
- Attendees will leave with strong practical knowledge of the issues and solutions for obtaining high-quality, sterile collections of targeted cells from a variety of sample types such as FFPE tissue, liquid biopsy, cell cultures, suspensions, and
more.
Speakers:
Karine Gousset, Ph.D.
Associate Professor of Virology, Dept. of Biology
California State University, Fresno
Isolation of Cellular Protrusions Using Laser Capture Microdissection for Subcellular Microproteomics
Studies
Laser Capture Microdissection (LCM) is uniquely tailored to specifically and reproducibly isolate and enrich individual cells based on morphology and/or using fluorescence markers. Numerous types of cellular protrusions have been described and the
majority of studies revolves around their possible functions. Tunneling nanotubes (TNTs), filopodia, growth cones or invadopodia are examples of cellular protrusions known to play a role in major cellular events such as cell differentiation, migration
and invasion. They also play critical role in diseases, including neurodegenerative diseases, virus spread and cancer. What has been missing so far is a method to analyze their individual protein composition and identify key differences within
these various subsets of cellular protrusions. Here, I will describe an LCM approach that we have developed to specifically isolate cellular protrusions and prepare samples for downstream microproteomics analyses. I will talk about sample preparation
for LCM and the best fixative method to preserve these cellular protrusions. I will also describe steps that we have taken to improve protein extraction from these samples for optimal mass spectrometry analysis, along with a brief note regarding
the type of mass spectrometers required for such small sample sizes.
Dr. Gousset obtained her PhD at UC Davis and did post-doctoral work at the Johns Hopkins University, NIH and the Pasteur Institute. She has a broad background in cellular biology, with specific training and expertise in microscopy, virology, molecular
biology techniques and microproteomics. The main focus of her laboratory is on the role of tunneling nanotubes (TNTs), a novel mechanism of functional connectivity between cells, in the spreading of viruses, misfolded protein aggregates (leading
to neurodegenerative diseases), as well as the role they may play in the proliferation and persistence of cancer.
Her lab has developed effective methods to identify the TNT proteome by exploring the limits of laser capture microdissection (LCM) and microproteomics and has overcome several major hurdles related to fixation and the small sample size of LCM, to
develop a robust approach to collect the TNT proteome (as well as the proteomes of other membrane protrusions, i.e., filopodia, axons, dendrites, and growth cones). Karine’s lab also developed a novel binary flow cytometry assay in which
to test identified proteins for their ability to affect TNT formation and function thereby streamlining the identification of functionally important TNT proteins from our microproteomic data.
Neil Umbreit, Ph.D.
Postdoctoral Fellow
Dana-Farber Cancer Institute
Single-Cell DNA Sequencing Reveals Mechanisms of Cancer Genome Evolution
Dr. Umbreit is a Research Fellow at Dana-Farber Cancer Institute and Harvard Medical School. His research focuses on how alterations to the genetic material in a cell contribute to the initiation and progression of cancers. He earned his Ph.D. in
Biochemistry at the University of Washington in 2014, and is the recipient of a Fellowship Award from the Damon-Runyon Cancer Research Foundation.
Yelena G. Golubeva, Ph.D.
Cancer Genomics Research Laboratory
Division of Cancer Epidemiology and Genetics, NCI, FNLCR, Leidos Biomedical Research
Target Enrichment of Archival Human Samples for Large Scale Studies in Cancer Epidemiology
Dr. Yelena Golubeva is Senior Scientist and Molecular pathology laboratory manager within the Cancer Genomic Research Laboratory, Division of Cancer Epidemiology and Genetics, NCI, FNLCR, Leidos Biomedical Research. She is a former scientist with
MedImmune, Inc and former leader of Laser Tissue Microdissection Core in Pathology-Histotechnology Laboratory of NCI-Frederick. Yelena has a Ph.D. in Entomology and Space and Aviation Medicine, and HT ASCP certification in histotechnology.
She has a vast experience in histological techniques and workflows, animal model genomic sample collection and preparation, immunostaining techniques and navigated hand microdissection of archival FFPE slides for molecular profiling studies. Dr. Golubeva
is the author of multiple method chapters on histological and genomic sample preparation, mouse embryo collection for genomic studies, IHC sample preparation, and Laser Capture Microdissection, and more than 30 peer-reviewed papers.
Cost: No cost!