3-dimensional imaging of biological structures by high resolution confocal scanning laser microscopy.
Imaging in confocal microscopy is characterized by the ability to make a selective image of just one plane inside a specimen, virtually unaffected -within certain limits- by the out-of-focus regions above and below it. This property, called optical sectioning, is accompanied by improved imaging transverse to the optical axis. We have coupled a confocal microscope to a computer system, making the combination of both an excellent instrument for mapping the 3-dimensional structure of extended specimens into a computer memory/data array. We measured that the volume element contributing to each data point has, under typical fluorescence conditions, a size of 0.2 X 0.2 X 0.72 micron. The data can be analysed and represented in various ways, i.e., stereoscopical views from any desired angle. After a description of the experimental arrangement, we show various examples of biological and food-structural studies. The microscope can be operated either in reflection or in fluorescence. In the latter mode a spectral element allows selection of the wavelength band of fluorescence light contributing to the image. In this way, we can distinguish various structures inside the cell and study their 3-dimensional relationships. Various applications in biology and the study of food structure are presented.
Brakenhoff
G J
GJ
Department of Electron Microscopy and Molecular Cytology, University of Amsterdam, The Netherlands.
van der Voort
H T
HT
van Spronsen
E A
EA
Nanninga
N
N
D000818:Animals; D002460:Cell Line; D004622:Embryo, Mammalian / Q000648:ultrastructure; D056890:Eukaryota / Q000648:ultrastructure; D005502:Food; D007834:Lasers; D008855:Microscopy, Electron, Scanning / Q000295:instrumentation* / Q000379:methods; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000648:ultrastructure; D013724:Teratoma