Real-time slit scanning microscopy in the meridional plane.
The standard microscope architecture around which confocal microscopes are built imposes fundamental restrictions on the speed with which images (three-dimensional data sets) can be obtained. Commercially available slit scanning confocal microscopes are able to produce optically sectioned images at frame rates well in excess of 100 Hz. However only the focal (x-y) plane can be imaged at this speed. To image a volume specimen it is necessary to physically change the distance between the objective lens and the specimen. This refocusing process is often necessarily slow and represents a bottleneck to the speed of image acquisition. We describe the construction of a slit scanning confocal microscope based on what we know to be a novel microscope architecture, which permits images of other planes and, particular, the meridional (x-z) plane to be acquired in real time.
Botcherby
Edward J
EJ
Department of Engineering Science, University of Oxford, Parks Road, Oxford OX1 3PJ, UK.
Booth
Martin J
MJ
Juskaitis
Rimas
R
Wilson
Tony
T
D005453:Fluorescence; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D011058:Pollen / Q000166:cytology; D013997:Time Factors