Book contents
- Frontmatter
- Contents
- Preface to the seventh edition
- List of contributors
- List of abbreviations
- 1 Basic principles
- 2 Cell culture techniques
- 3 Centrifugation
- 4 Microscopy
- 5 Molecular biology, bioinformatics and basic techniques
- 6 Recombinant DNA and genetic analysis
- 7 Immunochemical techniques
- 8 Protein structure, purification, characterisation and function analysis
- 9 Mass spectrometric techniques
- 10 Electrophoretic techniques
- 11 Chromatographic techniques
- 12 Spectroscopic techniques: I Spectrophotometric techniques
- 13 Spectroscopic techniques: II Structure and interactions
- 14 Radioisotope techniques
- 15 Enzymes
- 16 Principles of clinical biochemistry
- 17 Cell membrane receptors and cell signalling
- 18 Drug discovery and development
- Index
- Plate section
- References
This chapter is part of a book that is no longer available to purchase from Cambridge Core
4 - Microscopy
Book contents
- Frontmatter
- Contents
- Preface to the seventh edition
- List of contributors
- List of abbreviations
- 1 Basic principles
- 2 Cell culture techniques
- 3 Centrifugation
- 4 Microscopy
- 5 Molecular biology, bioinformatics and basic techniques
- 6 Recombinant DNA and genetic analysis
- 7 Immunochemical techniques
- 8 Protein structure, purification, characterisation and function analysis
- 9 Mass spectrometric techniques
- 10 Electrophoretic techniques
- 11 Chromatographic techniques
- 12 Spectroscopic techniques: I Spectrophotometric techniques
- 13 Spectroscopic techniques: II Structure and interactions
- 14 Radioisotope techniques
- 15 Enzymes
- 16 Principles of clinical biochemistry
- 17 Cell membrane receptors and cell signalling
- 18 Drug discovery and development
- Index
- Plate section
- References
Summary
INTRODUCTION
Biochemical analysis is frequently accompanied by microscopic examination of tissue, cell or organelle preparations. Such examinations are used in many different applications, for example: to evaluate the integrity of samples during an experiment; to map the fine details of the spatial distribution of macromolecules within cells; to directly measure biochemical events within living tissues.
There are two fundamentally different types of microscope: the light microscope and the electron microscope (Fig. 4.1). Light microscopes use a series of glass lenses to focus light in order to form an image whereas electron microscopes use electromagnetic lenses to focus a beam of electrons. Light microscopes are able to magnify to a maximum of approximately 1500 times whereas electron microscopes are capable of magnifying to a maximum of approximately 200 000 times.
Magnification is not the best measure of a microscope, however. Rather, resolution, the ability to distinguish between two closely spaced points in a specimen, is a much more reliable estimate of a microscope's utility. Standard light microscopes have a lateral resolution limit of about 0.5 micrometers (μm) for routine analysis. In contrast, electron microscopes have a lateral resolution of up to 1 nanometer (nm). Both living and dead specimens are viewed with a light microscope, and often in real colour, whereas only dead ones are viewed with an electron microscope, and never in real colour.
Information
- Type
- Chapter
- Information
- Principles and Techniques of Biochemistry and Molecular Biology , pp. 100 - 137Publisher: Cambridge University PressPrint publication year: 2010
References
(2003). Microscope Basics and Beyond. Melville, NY: Olympus of America. (Good well-illustrated primer on all aspects of basic light microscopy, also available online as a pdf. file.)Google Scholar
and (2004). Biological ultrastructure research: the first 50 years. Tissue Cell, 36, 83–94. (Ageless review of the early history of electron microscopy.)CrossRefGoogle ScholarPubMed
, , , , and (2007). Molecular Biology of the Cell, 5th edn. New York: Garland Science. (Basic introduction to all forms of microscopy and live cell imaging for the cell biologist.)Google Scholar
, and (2002). To 5D and beyond: quantitative fluorescence microscopy in the postgenomic era. Traffic, 3, 29–36. (Review of multidimensional imaging, methods of coping with large data sets and international image databases.)CrossRefGoogle ScholarPubMed
(2004). Mapping molecular landscapes inside cells. Biological Chemistry, 385, 865–872. (Review of electron tomography.)CrossRefGoogle ScholarPubMed
(2006). Optical Imaging Techniques in Cell Biology. Boca Raton, FL: CRC Press. (Overview of the entire field of light microscopy.)CrossRefGoogle Scholar
, , and (2006). Confocal quantification of cis-regulatory reporter gene expression in living sea urchins. Developmental Biology, 299, 543–550. (Practical example of quantitative measurements in living cells.)CrossRefGoogle Scholar
et al. (2009). Imaging transcription in living cells. Annual Review of Biophysics, 38, 173–196.CrossRefGoogle ScholarPubMed
and (2004). Cell motility under the microscope: Vorsprung durch Technik. Nature Reviews Molecular and Cell Biology, 5, 667–672. (Review of techniques used to study cell motility.)CrossRefGoogle ScholarPubMed
, and (eds.) (2009). Light Microscopy. Nature Milestones. www.nature.com/milestones/light-microscopy (Well-produced and complete review of all aspects of contemporary light microscopy.)
(2002). Envisioning Science: The Design and Craft of the Science Image. Cambridge, MA: MIT Press. (Popular work on imaging with some great tips and tricks for the stereomicroscope.)Google Scholar
, , and (2006). The fluorescent toolbox for assessing protein location and function. Science, 312, 217–224. (A review of the characteristics and benefits of using fluorescent probes to study proteins.)CrossRefGoogle ScholarPubMed
, , and (2003). Technicolor transgenics: imaging tools for functional genomics in the mouse. Nature Review Genetics, 4, 613–625.CrossRefGoogle Scholar
and (2009). Probing the macromolecular organisation of cells by electron tomography. Current Opinion in Cell Biology, 21, 89–96.CrossRefGoogle ScholarPubMed
and (1997). Video Microscopy: The Fundamentals, 2nd edn. New York: Plenum Press. (The classic text on live cell imaging, video microscopy and general microscopy.)CrossRefGoogle Scholar
and (2007). Imaging single events at the cell membrane. Nature Chemical Biology, 3, 92–98. (Overview of high resolution methods of light microscopy including TIRF.)CrossRefGoogle ScholarPubMed
, , and (2008). Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy. Sciencexpress, www.sciencexpress.org, 9 October 2008. (Application of scanning light microscopy to image living zebrafish embryos – stunning movies of zebrafish embryogenesis available online.)CrossRef
, , , , , , and (2008). SARS–Coronavirus replication is supported by a reticulovesicular network of modified endoplasmic reticulum. PLoS Biology, 6, 1957–1974. (Cryo-electron tomography in action.)CrossRefGoogle ScholarPubMed
and (2001). The neuronal naturalist: watching neurons in their native habitat. Nature Neuroscience (Suppl.), 4, 1215–1220.CrossRefGoogle ScholarPubMed
, , , , , , and (2007). Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system. Nature, 450, 56–62. (Imaginative use of reporter gene technology to label multiple neurons in living brains.)CrossRefGoogle Scholar
, , , and (2007). Three-dimensional imaging of Drosophila melanogaster. PLoS ONE, 2, E834. (Methods of three-dimensional imaging including confocal and optical projection tomography.)CrossRefGoogle ScholarPubMed
(2008). Scientific Imaging with PhotoShop: Methods, Measurement, and Output. Berkeley, CA: Pearson Education, Peachpit Press. (Practical manual on the use of PhotoShop for measuring and preparing images for publication.)Google Scholar
(2003). Practical Flow Cytometry, 4th edn. New York: John Wiley. (Wonderfully written book on basic fluorescence and flow cytometry.)CrossRefGoogle Scholar
and (2006). Basic Methods in Microscopy. Plainview, NY: Cold Spring Harbor Laboratory Press. (A good introduction to contemporary methods of imaging both fixed and living cells at both the light and electron microscope level.)Google Scholar
, and (2006). Modeling data across labs, genomes, space and time. Nature Cell Biology, 8, 1190–1194.CrossRefGoogle Scholar
, , and the OME Consortium (2009). Bioimage informatics for experimental biology. Annual Review of Biophysics, 38, 327–346.CrossRefGoogle ScholarPubMed
, , , , , , and (2007). Global analysis of gene expression during Drosophila embryogenesis. Genome Biology, 8, R145.CrossRefGoogle ScholarPubMed
and (2002). Imaging into the future: visualizing gene expression and protein interaction with fluorescent proteins. Nature Cell Biology, 4, E15–E20. (Good primer on GFP and FRET.)CrossRefGoogle Scholar
and (2008). FISH glossary: an overview of the fluorescence in situ hybridization technique. BioTechniques, 45, 385–409.CrossRefGoogle ScholarPubMed
, and (2001). Workingperson's guide to deconvolution in light microscopy. BioTechniques, 31, 1076–1097. (Comprehensive review of the deconvolution technique.)Google ScholarPubMed
, , , , and (2009). Advances in light microscopy for neuroscience. Annual Review of Neuroscience, 32, 435–506. (Complete coverage of all modern methods of imaging including super-resolution methods.)CrossRefGoogle ScholarPubMed
, , and (2002). Creating new fluorescent probes for cell biology. Nature Reviews Molecular Cell Biology, 3, 906–918. (Review of the development of fluorescent probes of biological activity especially reporter molecules.)CrossRefGoogle ScholarPubMed
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