Fluorescence Microscopy and Fluorescent Probes

✍ Scribed by Brian Herman (auth.), Jan Slavík (eds.)

• Publisher: Springer US
• Year: 1996
• Tongue: English
• Leaves: 292
• Edition: 1
• Category: Library

✦ Synopsis

Fluorescence microscopy images can be easily integrated into current video and computer image processing systems. People like visual observation; they like to watch a television or computer screen, and fluorescence techniques are thus becoming more and more popular. Since true in vivo experiments are simple to perform, samples can be directly seen and there is always the possibility of manipulating the samples during the experiments; it is an ideal technique for biology and medicine. Images are obtained by a classical (now called wide-field) fluorescence microscope, a confocal scanning microscope, upright or inverted, with epifluorescence or transmission. Computerized image processing may improve definition, and remove glare and scattered light signal. It also makes it possible to compute ratio images (ratio imaging both in excitation and in emission) or lifetime imaging. Image analysis programs may supply a great deal of additional data of various types, starting with calculations of the number of fluorescent objects, their shapes, brightness, etc. Fluorescence microscopy data may be complemented by classical measurement in the cuvette yr by flow cytometry.

✦ Table of Contents

Front Matter....Pages i-xviii
Fluorescence Microscopy: State of the Art....Pages 1-14
Fluorescence Lifetime-Resolved Imaging Microscopy: A General Description of Lifetime-Resolved Imaging Measurements....Pages 15-33
Confocal Fluorescence Lifetime Imaging....Pages 35-46
Multidimensional Fluorescence Microscopy: Optical Distortions in Quantitative Imaging of Biological Specimens....Pages 47-56
Fluorescent Probes....Pages 57-60
Flow Cytometry versus Fluorescence Microscopy....Pages 61-66
Multichannel Fluorescence Microscopy and Digital Imaging - On the Exciting Developments in Fluorescence Microscopy....Pages 67-70
Fluorescence Lifetime Imaging and Spectroscopy in Photobiology and Photomedicine....Pages 71-78
A Versatile Time-Resolved Laser Scanning Confocal Microscope....Pages 79-83
Disappearance of Cytoplasmic Ca 2+ Oscillations is a Sensitive Indicator of Photodamage in Pancreatic β-Cells....Pages 85-89
Distribution of Individual Cytoplasmic pH Values in a Cell Suspension....Pages 91-93
The Effect of Lysosomal pH on Lactoferrin-Dependent Iron Uptake in Tritrichomonas foetus ....Pages 95-99
On the Protein-Error of the Calcium-Sensitive Fluorescent Indicator Fura-Red....Pages 101-106
Cytoplasmic Ion Imaging: Evidence for Intracellular Calibration Heterogeneities of Ion-Sensitive Fluoroprobes....Pages 107-112
The Effect of Protein Binding on the Calibration Curve of the pH Indicator BCECF....Pages 113-118
Artifacts in Fluorescence Ratio Imaging....Pages 119-123
Use of Fluorescent Probes and CLSM for pH-Monitoring in the Whole Plant Tissue. pH-Changes in the Shoot Apex of Chenopodium rubrum Related to Organogenesis....Pages 125-132
Spatial Resolution of Cortical Cerebral Blood Flow and Brain Intracellular pH as Measured by In Vivo Fluorescence Imaging....Pages 133-138
Is a Potential-Sensitive Probe diS-C 3 (3) a Nernstian Dye?: Time-Resolved Fluorescence Study with Liposomes as a Model System....Pages 139-143
Kinetic Behavior of Potential-Sensitive Fluorescent Redistribution Probes: Modelling of the Time Course of Cell Staining....Pages 145-149
Speed of Accumulation of the Membrane Potential Indicator diS-C 3 (3) in Yeast Cells....Pages 151-155
Spectral Effects of Slow Dye Binding to Cells and Their Role in Membrane Potential Measurements....Pages 157-162
Exploitation of Rhodamine B in the Killer Toxin Research....Pages 163-167
“In Situ” Estimates of the Spatial Resolution for “Practical” Fluorescence Microscopy of Cell Nuclei....Pages 169-173
Requirements for a Computer-Based System for Fish Applications....Pages 175-178
Fluorescent Dyes and Dye Labelled Probes for Detection of Nucleic Acid Sequences in Biological Material....Pages 179-183
Fluorescence in Situ Hybridization (FISH) in Cytogenetics of Leukemia....Pages 185-189
Estimation of “START” in Saccharomyces cerevisiae by Flow Cytometry and Fluorescent Staining of DNA and Cell Protein....Pages 191-195
Fluorescence Image Cytometry of DNA Content: A Comparative Study of Three Fluorochromes and Four Fixation Protocols....Pages 197-201
In Vivo Tissue Characterization Using Environmentally Sensitive Fluorochromes....Pages 203-209
Sensitive and Rapid Detection of ß-Galactosidase Expression in Intact Cells by Microinjection of Fluorescent Substrate....Pages 211-215
Fluorogenic Substrates Reveal Genetic Differences in Aldehyde-Oxidating Enzyme Patterns in Rat Tissues....Pages 217-221
Binding of Prothrombin Fragment 1 to Phosphatidylserine Containing Vesicles: A Solvent Relaxation Study....Pages 223-227
New Thiol Active Fluorophores for Intracellular Thiols and Glutathione Measurement....Pages 229-233
Quantification of Macrophages in the Cardiovascular System of Hypercholesterolemic Rabbits by Use of Digital Image Processing....Pages 235-239
Fluorescence Assay for Studying P-Glycoprotein Function at Single Cell Level....Pages 241-245
Alterations of Vimentin-Nucleus Interactions as an Early Phase in Cholesterol Oxide - Induced Endothelial Cell Damage....Pages 247-251
Fluorescence Microscopy of Rye Cell Walls from Kernels to Incubated Doughs....Pages 253-255
Practical Approach for Immunohistochemical Staining of Muscle Biopsies....Pages 257-259
Rapid Automatic Segmentation of Fluorescent and Phase-Contrast Images of Bacteria....Pages 261-266
Use of Confocal Microscopy for Absolute Measurement of Cell Volume and Total Cell Surface Area....Pages 267-271
Cell Volume Measurements Using Confocal Laser Scanning Microscopy....Pages 273-277
Subcellular Cytofluorometry in Confocal Microscopy....Pages 279-283
Application of Confocal Microscopy to 3-D Reconstruction and Morphometrical Analysis of Capillaries....Pages 285-289
Retrieving Spatiotemporal Information from Confocal Data: A Study Using Melanotrope Cells of Xenopus Laevis ....Pages 291-295
Dynamics of Actin Measured by Fluorescence Correlation Microscopy (FCM)....Pages 297-301
Back Matter....Pages 303-306

✦ Subjects

Biological Microscopy;Animal Physiology;Biophysics and Biological Physics;Analytical Chemistry