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Microscopy Reference Library

Laser Scanning Confocal Microscopy (LSCM)

A majority of the literature pertaining to review articles on laser scanning confocal microscopy has been published in textbooks, edited article collections, and symposia, with only an intermittent sprinkling of papers in the scientific journals. The reviews listed in this section should be available to students and investigators who have access to subscriptions through their host institutions.

Wilhelm, S., Gröbler, B., Gluch, M. and Heinz, H.

Confocal laser scanning microscopy principles.  ZEISS Jena.  Written by scientists and engineers at ZEISS in Jena, Germany, this treatise on confocal microscopy reviews optical image formation and electronic signal processing. Included are discussions of the point spread function, resolution, digitization, and noise.

Amos, W. B. and White, J. G.

How the confocal laser scanning microscope entered biological research.  Biology of the Cell 95: 335-342 (2003).  An excellent historical review that describes the early development of laser scanning confocal microscopy. Key innovations are discussed, as well as detectors, instrument design, control software, and scanning systems. Also included are the early trials and tribulations with commercialization of the ideas.

Cox, G.

Biological confocal microscopy.  Materials Today 5: 34-41 (2002).  Targeted at casual readers, this review focuses on the basic concepts of confocal microscopy and related techniques in biology and medicine. The author discusses microscope configuration and provides several examples of applications using synthetic fluorophores and fluorescent proteins.

Webb, R. H.

Confocal optical microscopy.  Reports on Progress in Physics 59: 427-471 (1996).  An extensive review of laser scanning confocal microscopy describing the primary advantages and its practical implementation. Topics include a complete discussion of confocal theory, instrument design and construction, the point spread function, specimens, and general engineering details.

Paddock, S. W.

Principles and practices of laser scanning confocal microscopy.  Molecular Biotechnology 16: 127-149 (2000).  Expert confocal microscopist Stephen Paddock discusses the basic principles and evolution of the concept into modern sophisticated instruments. Included are principal imaging modes, optical sectioning, multi-color image acquisition, three-dimensional reconstruction, and live-cell imaging. Also discussed are specimen preparation, software issues, and troubleshooting.

Wright, S. J. and Wright, D. J.

Introduction to confocal microscopy.  Methods in Cell Biology 70: 1-85 (2002).  The authors present a comprehensive review of laser scanning confocal microscopy that discusses instrumental parameters, procedures and protocols for imaging fixed and live specimens, single and multiple labeling strategies, and optical sectioning. Also described are the presentation of confocal data and three-dimensional volume rendering.

Földes-Papp, Z., Demel, U. and Tilz, G. P.

Laser scanning confocal fluorescence microscopy: an overview.  International Immunopharmacology 3: 1715-1729 (2003).  As advertised, this article contains a general overview of laser scanning confocal microscopy, including discussions of the confocal principle, spatial resolution, optical sectioning, instrument diagrams, and lasers. Several examples of biological imaging using immunofluorescence are presented.

Halbhuber, K. J. and König, K.

Modern laser scanning microscopy in biology, biotechnology and medicine.  Annals of Anatomy 185: 1-20 (2003).  The authors present an extensive review of laser scanning confocal microscopy and closely related techniques with six pages of references. Included are numerous example images of tissue sections useful in anatomical research.

Zucker, R. M. and Price, O.

Evaluation of confocal microscopy system performance.  Cytometry Part A 44: 273-294 (2001).  The goal of this report is to develop quality assurance tests to ensure that the laser scanning confocal microscope is stable while delivering reproducible intensity measurements with excellent image quality. Tests to measure dichroic reflectivity, field illumination, lens performance, laser power output, spectral registration, and axial resolution, along with other critical factors were conducted.

Hepler, P. K. and Gunning, B. E. S.

Confocal Fluorescence microscopy of plant cells.  Protoplasma 201: 121-157 (1998).  This review addresses the contribution that laser scanning microscopy has made to the study of plant cells. Included are basic principles, image processing, laser illumination sources, and the use of fluorescent probes. Also discussed are investigations of the cytoskeleton, endomembranes, and nuclear components on both fixed and live plant cells.