Flow Cytometry and Laboratory Automation PDF

6/27/2024 Flow Cytometry and Laboratory Automation Preamble  PowerPoints DO NOT cover the details needed for the Unit exam  Unit Objectives are your study guide not this PowerPoint 1 ...

6/27/2024 Flow Cytometry and Laboratory Automation Preamble  PowerPoints DO NOT cover the details needed for the Unit exam  Unit Objectives are your study guide not this PowerPoint 1  6/27/2024 Chapter Overview Flow cytometry Immunoassay automation Instrumentation Validation Data acquisition and analysis Sample preparation Clinical applications Flow Cytometry Automated process that analyzes cells or beads in fluid suspensions for their light-scattering characteristics. Uses fluorochromes to identify cells or particles by size, shape, and antigenic properties. Allows for rapid and accurate detection of cells found in small numbers. 2  6/27/2024 Instrumentation 1. Fluidics  Allows for cell transport 2. Laser light source for cell illumination and identification:  Intrinsic parameters  Forward scatter (FSC)  Side scatter (SSC)  Extrinsic parameters 3. Optics and photodetectors  Signal detection 4. Computer  Data management Flow Cytometry Process  Cells are processed into a suspension, and the cytometer draws up the cell suspension and injects the sample inside a carrier stream of isotonic saline (sheath fluid) to form a laminar flow.  The sample stream is constrained by the carrier stream and is thus hydrodynamically focused so that the cells pass in a single file through the intersection of the laser light source  Each cell is interrogated by a light source that typically consists of one or more small air-cooled lasers. 3  6/27/2024 Flow Cytometry Process  Cells are labeled with a fluorochrome, or fluorescent molecule, that absorbs light across a spectrum of wavelengths and emits light of lower energy across a spectrum of longer wavelengths.  Each fluorochrome has a distinctive spectral pattern of absorption (excitation) and emission. Flow Cytometry and Laboratory Automation Light at two specific angles is measured by the flow cytometer: 1. Forward-angle light scatter (FSC) (