Image Cytometry
1st Edition
Published on 1. March 2001
Book
Paperback/Softback
116 pages
978-0-387-91618-7 (ISBN)
Description
Image cytometry is the measurement of such things as dimension, volume, shape, and reaction kinetics of biological and medical samples, This is an introductory practical guide to the use of image cytometry in biological and medical research, diagnosis and screening. The book describes the equipment necessary to do image cytometry (microscope, video camera, image software) and discusses the rapidly growing field of quantifying metabolic processes in cells. It clearly explains the various types of techniques available such as photometric, morphometric, planimetric and stereological methods. Extensive examples are used throughout in all the explanations. The book also helps the reader to select the correct procedures to use to obtain the required quantitative information.
The book will be very useful to anyone who uses the microscope in the laboratory and wishes to quantify tissue and cellular characteristics.
The book will be very useful to anyone who uses the microscope in the laboratory and wishes to quantify tissue and cellular characteristics.
More details
Series
Edition
2000
Language
English
Place of publication
Oxford
United States
Product notice
Paperback (trade)
Unsewn / adhesive bound
Illustrations
60
15 s/w Tabellen, 60 s/w Abbildungen
60 illus.
Dimensions
Height: 23.5 cm
Width: 15.5 cm
Thickness: 9 mm
Weight
259 gr
ISBN-13
978-0-387-91618-7 (9780387916187)
Schweitzer Classification
Content
Introduction.
1. Equipment.
Bright field and fluorescence microscope. Confocal microscopes. Objectives & relay lenses. White light illuminators. Fluorescent illuminators. filter & Monochromators. Autofocusing. Motorized stages. Imaging devices, signal formats & frame grabbers. Image cytometry software.
2. Calibration and Set Up.
Defining illumination needs. Optical path alignment. Linearization of camera response (bright & dark field, fluoro-luminescence). Noise reduction and sensor cooling. Signal integration. Spatial calibration. Selecting magnification to optimize contrast transfer and avoid diffraction problems. Object thickness and depth of field.
3. Transmitted light photometry.
Specimen preparation, product stoichiometry, standard calibration: cytochemistry. Immunochemistry. In situ hybridization. Autoradiography. Optical density and integrated optical density. Wavelength selection. Control for glare. Establishment of the reference unit (nuclei, cells, tissue area etc.). Single cell measurements. Tissue measurements. whole section measurements. Data analysis.
4. Emitted light photometry.
Specimen preparation and standard calibration. Fluorometry. Luminescence. High sensitive microscope systems (sensors, light sources, intensifiers). Multi-band pass filters and image shifts. Emission. Fading and antifading. Time-resolved fluorescence and luminescence. Single dye measurements in tissues and cells. Ploidy analysis by interphase cytogenetics. Ratio fluorescence measurements. Comparative Genomic Hybridization. Reaction kinetics in luminescence measurements. Metabolite mapping.
5. Labeling indexes and object counting.
Assessing a staining threshold for labeled objects. Planning a sampling design for tissue areas or isolated cells. Defining reference units: cells, section areas, parenchymal functional units. Manuals scoring and counting. Interactive stereological (point grid) methods. Interactive image processing methods. fully automated image analysis methods. Measuring a labeling index for positive nuclei, cytoplasm or membranes. Measuring the mitotic index. Source of variation, but off points and statistical treatment.
6. Morphometry.
Areas, shapes, distances, profiles and contours. Isolated cells and tissue sections. Stereological methods. Image analysis methods. Three dimensional reconstruction of tissue structures (part of cytometry??). Real time morphometry on living cells. Measuring form factors in nuclear grading. Measuring AgNORs. Measuring endothelial area and microvessel density. Measuring area fraction of tissues. Statistical treatment.
7. Quality assurance issues.
8. Advance cytometric systems in the clinical setting.
Automatic PAP-test screeners. Laser scanning cytometers. Chromosomes and karyotype analyzers.
1. Equipment.
Bright field and fluorescence microscope. Confocal microscopes. Objectives & relay lenses. White light illuminators. Fluorescent illuminators. filter & Monochromators. Autofocusing. Motorized stages. Imaging devices, signal formats & frame grabbers. Image cytometry software.
2. Calibration and Set Up.
Defining illumination needs. Optical path alignment. Linearization of camera response (bright & dark field, fluoro-luminescence). Noise reduction and sensor cooling. Signal integration. Spatial calibration. Selecting magnification to optimize contrast transfer and avoid diffraction problems. Object thickness and depth of field.
3. Transmitted light photometry.
Specimen preparation, product stoichiometry, standard calibration: cytochemistry. Immunochemistry. In situ hybridization. Autoradiography. Optical density and integrated optical density. Wavelength selection. Control for glare. Establishment of the reference unit (nuclei, cells, tissue area etc.). Single cell measurements. Tissue measurements. whole section measurements. Data analysis.
4. Emitted light photometry.
Specimen preparation and standard calibration. Fluorometry. Luminescence. High sensitive microscope systems (sensors, light sources, intensifiers). Multi-band pass filters and image shifts. Emission. Fading and antifading. Time-resolved fluorescence and luminescence. Single dye measurements in tissues and cells. Ploidy analysis by interphase cytogenetics. Ratio fluorescence measurements. Comparative Genomic Hybridization. Reaction kinetics in luminescence measurements. Metabolite mapping.
5. Labeling indexes and object counting.
Assessing a staining threshold for labeled objects. Planning a sampling design for tissue areas or isolated cells. Defining reference units: cells, section areas, parenchymal functional units. Manuals scoring and counting. Interactive stereological (point grid) methods. Interactive image processing methods. fully automated image analysis methods. Measuring a labeling index for positive nuclei, cytoplasm or membranes. Measuring the mitotic index. Source of variation, but off points and statistical treatment.
6. Morphometry.
Areas, shapes, distances, profiles and contours. Isolated cells and tissue sections. Stereological methods. Image analysis methods. Three dimensional reconstruction of tissue structures (part of cytometry??). Real time morphometry on living cells. Measuring form factors in nuclear grading. Measuring AgNORs. Measuring endothelial area and microvessel density. Measuring area fraction of tissues. Statistical treatment.
7. Quality assurance issues.
8. Advance cytometric systems in the clinical setting.
Automatic PAP-test screeners. Laser scanning cytometers. Chromosomes and karyotype analyzers.