4.3 Review

Modern laser scanning microscopy in biology, biotechnology and medicine

Journal

ANNALS OF ANATOMY-ANATOMISCHER ANZEIGER
Volume 185, Issue 1, Pages 1-20

Publisher

ELSEVIER GMBH, URBAN & FISCHER VERLAG
DOI: 10.1016/S0940-9602(03)80002-X

Keywords

confocal laser scanning microscopy; multiphoton laser scanning microscopy; oxygen radicals; alkaline phosphatase; peroxidase; Jenfluor((R))ap; Jenfluor((R))px Red; Jenchrom((R))px Blue

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Laser microscopic techniques currently used in morphology and cell biology represent highly sensitive tools for detecting biomolecules within their natural environment. Use of the fluorescence-, reflectance- and transmission modes of confocal laser scanning microscopes (CLSM) equipped with He-Ne- and Ar+-ion lasers for Ce-IV and DAB based detection of endogenous or immunobound enzymatic activities in tissue sections (vibratome, cryostat, paraffin and semithin plastic sections) opens a wide range of interesting new possibilities in cellular and molecular biology. Increased resolution power, blur-free confocal imaging, higher sensitivity, optical sectioning capability and 3D-image analysis provide a large quantity of valuable information about biological objects specimens. The new infrared multiphoton laser scanning microscopy (NIR-LSM) is increasingly becoming the optical tool of choice for (a) fluorescence imaging of cellular and subcellular components with high spatial and temporal resolution, (b) fluorescence resonance energy transfer between physiologically relevant molecular species involving protein-protein interactions, (c) nanoprocessing within living cells and tissues, with varied applications in (d) photochemistry and (e) medical diagnostics as well. Both, CLSM and NIR-LSM as modern microscopical strategies are indispensable in basic research and will prove to be invaluable for clinical diagnostic studies and therapy in the near future.

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