A high-resolution, confocal laser-scanning microscope and flash photolysis system for physiological studies.

Abstract:

:We describe the construction of a high-resolution confocal laser-scanning microscope, and illustrate its use for studying elementary Ca2+ signalling events in cells. An avalanche photodiode module and simple optical path provide a high efficiency system for detection of fluorescence signals, allowing use of a small confocal aperture giving near diffraction-limited spatial resolution (< 300 nm lateral and < 400 nm axial). When operated in line-scan mode, the maximum temporal resolution is 1 ms, and the associated computer software allows complete flexibility to record line-scans continuously for long (minutes) periods or to obtain any desired pixel resolution in x-y scans. An independent UV irradiation system permits simultaneous photolysis of caged compounds over either a uniform, wide field (arc lamp source) or at a tightly focussed spot (frequency-tripled Nd:YAG laser). The microscope thus provides a versatile tool for optical studies of dynamic cellular processes, as well as excellent resolution for morphological studies. The confocal scanner can be added to virtually any inverted microscope for a component cost that is only a small fraction of that of comparable commercial instruments, yet offers better performance and greater versatility.

journal_name

Cell Calcium

journal_title

Cell calcium

authors

Parker I,Callamaras N,Wier WG

doi

10.1016/s0143-4160(97)90055-5

subject

Has Abstract

pub_date

1997-06-01 00:00:00

pages

441-52

issue

6

eissn

0143-4160

issn

1532-1991

pii

S0143-4160(97)90055-5

journal_volume

21

pub_type

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