FlowCytometry Basics

Flow Cytometry Anna Merca - Philippe Melas - Tolga Sütlü Stockholm Research School in Molecular Life Sciences 14 October 2005

Outline ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Introduction ,[object Object],[object Object],[object Object],[object Object]

How does it work? ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Direct beam stop Laser Light High angle scatter : (Side Scatter) Cell structure Low angle scatter : (Forward Scatter) Cell size Fluorescence at longer wavelengths Flow cell Direction of flow

Basic Optics of a Flow Cytometer ( An automated fluorescent microscope) Dichroic mirrors 1 2 3 Laser(s) Cell Collection Lenses Scatter Low & High angle Photomultiplier tubes

Analysis Applications ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Analyzing the graph -one color- CD3 NUMBER CD4 NUMBER CD8 NUMBER

Analyzing the graph CD4 0 1 10 2 10 3 10 4 0 10 CD3 CD3 CD4 CD3+CD4+ green CD3-CD4+ cyan CD3+CD4- cyan CD3-CD4- black

Apoptosis vs. Necrosis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

How to study apoptosis: Caspases ,[object Object]

How to study apoptosis: Mitochondrial Membrane Potential ,[object Object]

How to study apoptosis: Plasma Membrane ,[object Object]

Cell Sorting 1 ,[object Object],[object Object],[object Object],[object Object]

Cell Sorting 2 Charged Plates Collection Tubes

Benefits of Flow Cytometry ,[object Object],[object Object],[object Object],[object Object]

Limitations ,[object Object],[object Object],[object Object],[object Object],[object Object]

FlowCytometry Basics

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