Optical modifications enabling simultaneous confocal imaging with dyes excited by ultraviolet- and visible-wavelength light

C. Bliton, James D Lechleiter, D. E. Clapham

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Optical modifications to a confocal scanning laser microscope are described which allow simultaneous fluorescence imaging of living specimens excited by ultraviolet (UV)- and visible-wavelength light. Modifications to a Bio-Rad MRC 600 Lasersharp confocal microscope include the introduction of UV-path-specific lenses and a specially designed UV transmitting eyepiece and tube lens. Upon UV excitation these modifications provide similar similar resolution and field flatness when compared with visible confocal microscopy. The UV-path-specific optics could be adjusted to correct for varying amounts of longitudinal chromatic aberration in commericially available objectives. Eyepiece and tube lenses were chromatically corrected for UV through visible wavelengths to minimize lateral chromatic error. With these modifications, UV-wavelength light may be used to excitate ratioing dyes to quantify intracellular ion concentrations, or as an energy source to release caged compounds in a spatially restricted volume, while simultaneously imaging with dyes excitated by visible-wavelength light.

Original languageEnglish (US)
Pages (from-to)15-26
Number of pages12
JournalJournal of Microscopy
Volume169
Issue number1
StatePublished - 1993
Externally publishedYes

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Lenses
Coloring Agents
Dyes
dyes
eyepieces
Imaging techniques
Light
Wavelength
lenses
Color
wavelengths
Microscopes
Optical Imaging
microscopes
Ultraviolet Rays
tubes
Confocal Microscopy
Confocal microscopy
Lasers
flatness

ASJC Scopus subject areas

  • Instrumentation

Cite this

Optical modifications enabling simultaneous confocal imaging with dyes excited by ultraviolet- and visible-wavelength light. / Bliton, C.; Lechleiter, James D; Clapham, D. E.

In: Journal of Microscopy, Vol. 169, No. 1, 1993, p. 15-26.

Research output: Contribution to journalArticle

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