Confocal Laser Scanning Microscopy There are two major forms of laser scanning microscopy: confocal laser scanning microscopy (CLSM) and multiphoton laser scanning microscopy (MPLSM). Information on: X-t scans and X-Y scans; Confocal Laser Scanning Microscopy; Multiphoton Laser Scanning Microscopy; MPLSM requires no pin hole; Advantages of MPLSM over CLSM.
Confocal Microscopy Specimen Preparation Using Synthetic Fluorophores and Immunofluorescence Includes staining protocols: Triple-Staining Adherent Cells with MitoTracker, Phalloidin,& Nuclear Dyes; Staining Adherent Cells with Cytokeratin Primary Antibodies & Synthetic Fluorophores; Staining Adherent Cells with Intermediate Filament Primary Antibodies & Synthetic Fluorophores; Staining Cells & Tissue Cryosections with Tubulin Primary Antibodies, Phallotoxins,& Synthetic Fluorophores; Triple-Staining Tissue Cryosections with Wheat Germ Agglutinin, Phalloidin,& Nuclear Dyes; etc.
Confocal Microscopy: Speciman Preparation and Imaging Basic information on confocal microscopy, includes: Specimen Preparation and Imaging; Objective Lens Parameters and Optical Section Thickness; The Objective Lens; Probes for Confocal Imaging; Autofluorescence; Collecting Images; Troubleshooting; Image Processing and Publication;
Information on Confocal Imaging Information on: Applications of Confocal Microscopy; Practical Instruments; Limitations of point-scanning Confocal Microscopy; Parallel beam confocal Imaging Systems.
Live Cell Spinning Disk Confocal Fluore Imaging of Cells- Colocalization of Fluorescent Protein Tags Protocol describes the acquisition and processing of confocal
fluorescent and bright field images of live cells, expressing cyan fluorescent protein(CFP) and/or yellow fluorescent protein (YFP), with a spinning disk confocal head on a Zeiss Axiovert 200 M microscope. This procedure is used to help determine if N- or Cterminal tagging of signaling molecules alters the steady state localization pattern of the signaling protein in question.
Live Cell Spinning Disk Confocal Fluorescence Imaging of Cells- YFP & Bright Field—Three Z Axis Protocol describes the acquisition and processing of confocal
fluorescent and bright field images of live cells expressing yellow fluorescent protein (YFP), with a spinning disk confocal head on a Zeiss Axiovert 200 M microscope when three planes along the z-axis of the cell are acquired. Protocol includes: Description of Microscope and Imaging Setup; Description of Acquisition Parameters; Image Processing.
Looking Inside Cells and Tissues by Optical Sectioning with a Confocal Laser Scanning Microscope Confocal laser scanning microscopy (CLSM) is a relatively new light microscopical imaging technique which has found wide applications in the biological sciences. The primary value of the CLSM to the biologist is its ability to produce optical sections through a 3-D specimen-e.g., an entire cell or a piece of tissue - that, to a good approximation, contain information from only one focal plane. Article includes principle and applications of confocal laser scanning microscope.
Protocol for Live Cell Spinning Disk Confocal Fluorescence Imaging of Cells on a Zeiss Protocol describes the acquisition of confocal fluorescent and
bright field images of live cells, expressing cyan fluorescent protein (CFP) and/or yellow fluorescent protein (YFP), with a spinning disk confocal head on a Zeiss Axiovert 200 M
microscope. Protocol includes: Description of Microscope and Imaging Setup; Description of Acquisition Parameters; Image Processing; Movie Processing.
Protocol for Nuclear Staining of Plants for Confocal Microscopy Protocol describes a useful way to observe the development of embryos, as well as meristems & young primordia developing at the shoot apex by confocal microscopy after staining the nuclei with propidium iodide. The number of cells can be exactly quantified in a meristem or in young primordia. Because embryonic & meristematic cells are largely filled out by their nuclei, it is easier to image only the nuclei. This method allows analysis of whole-mount material, which is more easily reconstructed.