Research Report 2010 - MDC

Structure of the GroupGroup LeaderHans-Peter RahnStudent AssistantJens HartwigSecretariatDaniela KeynerStart of group: January 2009Hans-Peter RahnPreparative FlowcytometryFluorescence-activated cell sorting (FACS) is a specialized type of flowcytometry. It wasdesigned in 1971 by Len Herzenberg to provide a fast physical separation method forfractionating a heterogeneous mixtures of cells into distinct subsets, based upon the lightscattering and fluorescent characteristics of different cell types.The FACS Core Facility assists researchers at the MDC with two state-of-the-art, digital highspeed“FACSAria” sorters and one analog high-speed “FACSVantage SE” sorter from BD. Thefacility has been used by more than 30 scientific groups at the MDC. Sorted cells are oftenused for microarray- or sequencing-based gene expression analysis, quantitative real-timePCR, DNA sequencing, live cell imaging or adoptive transfer experiments in animals.Recent Projects:Sophisticated FACS applications are used in theresearch group of Frank Rosenbauer, with the theme“Cancer, Stem Cells, and Transcription Factors”. Theinterest of this group is to integrate hematopoitic stemcell characteristics with underlying genetic and epigeneticregulatory mechanisms. To identify specific differentiationstates of hematopoetic and leukemic cells, thegroup developed new staining methods using up tonine colors. Routinely, seven color sorts of mouse bonemarrow cells are used to analyse hematopoetic cells invivo and in vitro.The “Molecular Tumor Genetics and Immunogenetics”group of Martin Lipp investigates the function of T- andB- cells during the development of acute and chronicinflammation. The group focuses on distinct memoryand effector T cell populations which are important forthe development of antibody-based adaptive immuneresponses. In this connection central memory, effector/memoryund follicular B helper T cells are enrichedby FACS using specific patterns of surface markers onthese cells.The “Hematoloy, Oncology and Tumorimmunology”group of Bernd Dörken “Hematoloy, Oncology andTumorimmunology” uses FACS enrichment of tumorand primary murine hematopoetic cells in order toanalyse the function of ectopically expressed genes incomparison to the endogenous expression levels inlymphoma cells.Together with the research group of Manfred Gossen,FACS was used to create long term stable transgeniccell clones, without the influence of additional anitbioticresistance genes.To study early embryogenesis employing high-throughputgenomics as well as biochemistry assays, the“Systems Biology” lab of Nikolaus Rajewsky devised anew method to collect large amounts of preciselystaged C. elegans embryos by FACS. This method willmake a contribution towards a more complete understandingof gene regulatory networks during early C.elegans development.226 Technology Platforms