In situ fluorescence imaging of organs through compact scanning head for confocal laser microscopy

Taisuke Ota; Hiroya Fukuyama; Yasuhige Ishihara; Hideo Tanaka; Tetsuro Takamatsu

doi:10.1117/1.1890411

1 March 2005 In situ fluorescence imaging of organs through compact scanning head for confocal laser microscopy

Taisuke Ota, Hiroya Fukuyama, Yasuhige Ishihara, Hideo Tanaka, Tetsuro Takamatsu

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Journal of Biomedical Optics, Vol. 10, Issue 2, 024010 (March 2005). https://doi.org/10.1117/1.1890411

Abstract

We develop a compact scanning head for use in laser confocal fluorescence microscopy for in situ fluorescence imaging of organs. The head, cylindrical in shape, has 3.5 mm diameter and 30 mm length, and is thus small enough to operate in a living rat heart. The lateral and axial resolutions, defined as full widths at half maximum (FWHM) of a point spread function (PSF), measures 1.0 and 5.0 µm, respectively, for 488-nm excitation and 1.0 and 5.4 µm, respectively, for 543-nm excitation. The chromatic aberration between 488- and 543-nm laser beams is well suppressed. We perform Ca²⁺ imaging in cardiomyocytes through the right ventricular chamber of a perfused rat heart in line-scan mode with 2.9-ms time resolution. We also carried out two-color imaging of a fixed mouse heart and liver with subcellular resolution. The compact head of the microscope equipped with a line-scan imaging mode and two-color imaging mode is useful for in situ imaging in living organs with subcellular resolution and can advantageously be applied to in vivo research.

©(2005) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Taisuke Ota, Hiroya Fukuyama, Yasuhige Ishihara, Hideo Tanaka, and Tetsuro Takamatsu "In situ fluorescence imaging of organs through compact scanning head for confocal laser microscopy," Journal of Biomedical Optics 10(2), 024010 (1 March 2005). https://doi.org/10.1117/1.1890411

Published: 1 March 2005

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KEYWORDS

Head

Heart

Calcium

Confocal microscopy

Luminescence

Image resolution

Microscopes

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