Title: Flow Cytometry Basic Training 1 Flow Cytometry Basic Training • A Look Inside the Box James MarvinFlow Cytometry Facility Northwestern University 2 (No Transcript) 3 Section I • Background Information on Flow Cytometry 4 The Many Parts of Flow • Experimental design • Sample preparation • Choosing the proper instrument • Setting up the instrument • Collecting the proper data • Interpreting the data • Graphics presentation and publication • Sorting Specific Applications Courses Flow Basics Data Analysis 5 What Is Flow Cytometry? • Flow cells in motion • Cyto cell • Metry measure • Measuring properties of cells while in a fluid stream 6 (No Transcript) 7 Cytometry vs. Flow Cytometry • Cytometry • Localization of antigen is possible • Poor enumeration of cell subtypes • Limiting number of simultaneous measurements • Flow Cytometry. • Cannot tell you where antigen is.

Feb 4, 2004 - From the ‡Tumour Targeting Program, Ludwig Institute for Cancer Research, Austin Hospital. Cently published crystal structures of the EGFR (29–31), we. Cells were then induced for protein display by transfer to minimal media. Cytometer (Beckman Coulter) and analyzed with WinMDI cytometry.

• Can analyze many cells in a short time frame. • Can look at numerous parameters at once.

8 Uses of Flow Cytometry • It can be used for • Immunophenotyping • DNA cell cycle/tumor ploidy • Membrane potential • Ion flux • Cell viability • Intracellular protein staining • pH changes • Cell tracking and proliferation • Sorting • Redox state • Chromatin structure • Total protein • Lipids • Surface charge • Membrane fusion/runover • Enzyme activity • Oxidative metabolism • Sulfhydryl groups/glutathione 9 Background Info Summary • Flow Cytometry is a quickly expanding technology • Has continually increased in popularity since the mid 1980s. • Gives us the ability to analyze many properties of many cells in very little time 10 Section II • The 4 Main Components of a Flow Cytometer 11 What Happens in a Flow Cytometer? • Cells in suspension flow single file past • a focused laser where they scatter light and emit fluorescence that is filtered and collected • then converted to digitized values that are stored in a file • Which can then be read by specialized software. 12 What Happens in a Flow Cytometer (Simplified) 13 The Fluidics SystemCells in suspension flow single file • You need to have the cells flow one-by-one into the cytometer to do single cell analysis • Accomplished through a pressurized laminar flow system. • The sample is injected into a sheath fluid as it passes through a small orifice (50um-300um) 14 Fluidics Schematic Sample Tube 15 How The Flow Cell Works • The cells from the sample tube are injected into the sheath stream • Flow in a flow cell is laminar. Sengoku basara 4 pc bitmoji for android phone. • Hydrodynamic focusing pushes the cells to line up single file along their long axis.

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