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High‐resolution confocal microscopy using synchrotron radiation
Author(s) -
VAN DER OORD C. J. R.,
JONES G. R.,
SHAW D. A.,
MUNRO I. H.,
LEVINE Y. K.,
GERRITSEN H. C.
Publication year - 1996
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 - 10.1046/j.1365-2818.1996.66432.x
Subject(s) - synchrotron radiation , optics , microscope , achromatic lens , confocal , microscopy , synchrotron , materials science , resolution (logic) , confocal microscopy , optoelectronics , chemistry , physics , artificial intelligence , computer science
A confocal scanning light microscope coupled to the Daresbury Synchrotron Radiation Source is described. The broad spectrum of synchrotron radiation and the application of achromatic quartz/CaF 2 optics allows for confocal imaging over the wavelength range 200–700 nm. This includes UV light, which is particularly suitable for high‐resolution imaging. The results of test measurements using 290‐nm light indicate that a lateral resolution better than 100 nm is obtained. An additional advantage of the white synchrotron radiation is that the excitation wavelength can be chosen to match the absorption band of any fluorescent dye. The availability of UV light for confocal microscopy enables studies of naturally occurring fluorophores. The potential applications of the microscope are illustrated by the real‐time imaging of hormone traffic using the naturally occurring oestrogen coumestrol. (The IUPAC name for coumestrol is 3,9‐dihydroxy‐6H‐benzofuro[3,2‐c][1]benzopyran‐6‐one ( Chem. Abstr. Reg. No . 479‐13‐0). The trivial name will be used throughout this paper.)