Premium
A confocal laser microscope scanner for digital recording of optical serial sections
Author(s) -
Carlsson K.,
Liljeborg A.
Publication year - 1989
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1989.tb00557.x
Subject(s) - microscope , laser scanning , optics , scanner , confocal , raster scan , optical sectioning , materials science , laser , pinhole (optics) , stereo microscope , physics
SUMMARY A confocal laser microscope scanner developed at our institute is described. Since an ordinary microscope is used, it is easy to view the specimen prior to scanning. Confocal imaging is obtained by laser spot illumination, and by focusing the reflected or fluorescent light from the specimen onto a pinhole aperture in front of the detector (a photomultiplier tube). Two rotating mirrors are used to scan the laser beam in a raster pattern. The scanner is controlled by a microprocessor which coordinates scanning, data display, and data transfer to a host computer equipped with an array processor. Digital images with up to 1024 × 1024 pixels and 256 grey levels can be recorded. The optical sectioning property of confocal scanning is used to record thin (∼ 1 μm) sections of a specimen without the need for mechanical sectioning. By using computer‐control to adjust the focus of the microscope, a stack of consecutive sections can be automatically recorded. A computer is then used to display the 3‐D structure of the specimen. It is also possible to obtain quantitative information, both geometric and photometric. In addition to confocal laser scanning, it is easy to perform non‐confocal laser scanning, or to use conventional microscopic illumination techniques for (non‐confocal) scanning. The design has proved reliable and stable, requiring very few adjustments and realignments. Results obtained with this scanner are reported, and some limitations of the technique are discussed.