Foundations of Confocal Scanned Imaging in Light Microscopy -- Fundamental Limits in Confocal Microscopy -- Special Optical Elements -- Points, Pixels, and Gray Levels: Digitizing Image Data -- Laser Sources for Confocal Microscopy -- Non-Laser Light Sources for Three-Dimensional Microscopy -- Objective Lenses for Confocal Microscopy -- The Contrast Formation in Optical Microscopy -- The Intermediate Optical System of Laser-Scanning Confocal Microscopes -- Disk-Scanning Confocal Microscopy -- Measuring the Real Point Spread Function of High Numerical Aperture Microscope Objective Lenses -- Photon Detectors for Confocal Microscopy -- Structured Illumination Methods -- Visualization Systems for Multi-Dimensional Microscopy Images -- Automated Three-Dimensional Image Analysis Methods for Confocal Microscopy -- Fluorophores for Confocal Microscopy: Photophysics and Photochemistry -- Practical Considerations in the Selection and Application of Fluorescent Probes -- Guiding Principles of Specimen Preservation for Confocal Fluorescence Microscopy -- Confocal Microscopy of Living Cells -- Aberrations in Confocal and Multi-Photon Fluorescence Microscopy Induced by Refractive Index Mismatch -- Interaction of Light with Botanical Specimens -- Signal-to-Noise Ratio in Confocal Microscopes -- Comparison of Widefield/Deconvolution and Confocal Microscopy for Three-Dimensional Imaging -- Blind Deconvolution -- Image Enhancement by Deconvolution -- Fiber-Optics in Scanning Optical Microscopy -- Fluorescence Lifetime Imaging in Scanning Microscopy -- Multi-Photon Molecular Excitation in Laser-Scanning Microscopy -- Multifocal Multi-Photon Microscopy -- 4Pi Microscopy -- Nanoscale Resolution with Focused Light: Stimulated Emission Depletion and Other Reversible Saturable Optical Fluorescence Transitions Microscopy Concepts -- Mass Storage, Display, and Hard Copy -- Coherent Anti-Stokes Raman Scattering Microscopy -- Related Methods for Three-Dimensional Imaging -- Tutorial on Practical Confocal Microscopy and Use of the Confocal Test Specimen -- Practical Confocal Microscopy -- Selective Plane Illumination Microscopy -- Cell Damage During Multi-Photon Microscopy -- Photobleaching -- Nonlinear (Harmonic Generation) Optical Microscopy -- Imaging Brain Slices -- Fluorescent Ion Measurement -- Confocal and Multi-Photon Imaging of Living Embryos -- Imaging Plant Cells -- Practical Fluorescence Resonance Energy Transfer or Molecular Nanobioscopy of Living Cells -- Automated Confocal Imaging and High-Content Screening for Cytomics -- Automated Interpretation of Subcellular Location Patterns from Three-Dimensional Confocal Microscopy -- Display and Presentation Software -- When Light Microscope Resolution Is Not Enough:Correlational Light Microscopy and Electron Microscopy -- Databases for Two- and Three-Dimensional Microscopical Images in Biology -- Confocal Microscopy of Biofilms — Spatiotemporal Approaches -- Bibliography of Confocal Microscopy.

Handbook of biological confocal microscopy

Fundamental and practical limits in confocal light microscopy

The revival of light microscope techniques and instrumentation has had major impact on biological studies of the mechanism of cell movements. This paper surnarizes some of the contributions of the participants in this session to this important area of biological science and gives a brief literature review of their past work.

Computer Aided Microscopy And Image Processing: An Overview Of Studies On Cell Movement.

The previous chapter described how the confocal approach developed from conventional light microscopy and outlined the basic advantages gained by the use of confocal sampling techniques, primarily that the exclusion of light by the confocal pinhole makes it possible to record data from optical sections. This chapter will discuss the fundamental considerations that limit the performance of all confocal microscopes. Though at present no commercially available equipment approaches these limits, some simple tests will be described to help the user assess how well a given instrument performs. Additional information to help the user to operate the confocal microscope in an optimal manner can be found in Chapter 35, “ATutorial on Confocal Microscopy,” and Chapter 36, “Practical Confocal Microscopy.” These also include methods for measuring resolution and other useful parameters.

Fundamental Limits in Confocal Microscopy

Nanometer-scale motion analysis of microtubule-based motor enzymes

The recent application of digital imaging technologies to the analysis of video images from a DIC (differential interference contrast) light microscope has given 1-2 nm precision for the measurement of movements of submicron particles at 30 Hz. [Gelles et al., Nature 331(1988)450]. This technique has been applied to the study of the movements of 1) motor proteins (kinesin and cytoplasmic dynein) on microtubules (MTs) and 2) membrane glycoproteins in cellular plasma membranes. In both cases the nanometer measurements have provided important new insights into the molecular mechanisms which govern those biological movements.

Nanometer Measurements Of Motor Protein And Membrane Glycoprotein Movements

Analyzing kinesin and cytoplasmic-dynein driven movement with nanometer precision

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Nanometer-scale motion analysis of microtubule-based motor enzymes

Nanometer Measurements Of Motor Protein And Membrane Glycoprotein Movements

Analyzing kinesin and cytoplasmic-dynein driven movement with nanometer precision