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Imaging has become a vital tool for researchers in all aspects of biology. Recent advances in microscope technology, labeling techniques and gene and protein manipulation methods have led to breakthroughs in our understanding of biological processes. In order to take advantage of these techniques, biologists need to understand the fundamental techniques of microscopy. The methods found here, drawn from the popular laboratory standard manual Cells: A Laboratory Manual,
provide a solid course in the basics of using the microscope in a biology laboratory.
Basic Methods in Microscopy provides an essential guide to light microscopy, fluorescence microscopy, confocal microscopy, multiphoton microscopy and electron microscopy, preparation of tissues and cells, labeling of specimens and analysis of cellular events.
This manual is an important tool for any biology researcher employing imaging as a research method.
- 1. Light Microscopy
- 2. Confocal Microscopy, Deconvolution, and Structured Illumination Methods
- 3. Multiphoton and Multispectral Laserscanning Microscopy
- 4. Preparation of Cells and Tissues for Fluorescence Microscopy
- 5. Nonimmunological Fluorescent Labeling of Cellular Structures
- 6. Introduction to Immunofluorescence Microscopy
- 7. Immunostaining of Microtubules, Microtubuleassociated Proteins, and Intermediate Filaments
- 8. Immunofluorescence Localization of Actin
- 9. Immunofluorescence Localization of Nuclear Proteins
- 10. Immunofluorescence Methods for Saccharomyces cerevisiae
- 11. Immunofluorescence Methods for Drosophila Tissues
- 12. Immunofluorescence Methods for Caenorhabditis elegans
- 13. Analyzing DNA Replication: Nonisotopic Labeling
- 14. Analyzing RNA Synthesis: Nonisotopic Labeling
- 15. Fluorescence In Situ Hybridization to DNA
- 16. WholeMount Fluorescence In Situ Hybridization to Drosophila Chromosomal DNA
- 17. In Situ Hybridization to RNA
- 18. WholeMount In Situ Detection of RNAs in Vertebrate Embryos and Isolated Organs
- 19. Image Production Using Transmission Electron Microscopy
- 20. Preparative Methods for Transmission Electron Microscopy
- 21. Immunoelectron Microscopy
- APPENDIX 1. Microscopy: Lenses, Filters, and Emission/Excitation Spectra
- APPENDIX 2. Cautions
In summary, this book is a useful reference for a teaching laboratory or a microscopy facility with very general interests in a variety of microscopy techniques primarily for fixed tissues.
Any research laboratory that uses microscopy would benefit immensely by having this volume on hand. The components, from underlying optical principles and theories behind the most basic light microscope to the highly advanced transmission electron microscope, are clearly described in the text, figures, and legends. A variety of techniques in fluorescence microscopy are presented, including protocols for the preparation of specimens, preparation of slides and cover slips, and proper care, maintenance, and cleaning of optical equipment. A handy troubleshooting section provides guidance on how to correct imaging problems.
The Quarterly Review of Biology