A new fluorescence microscopy for tomographic observation of microcirculation by using dual-beam slit laser illumination

Masahiro Shibata, Takashi Kawamura, Mitra Sohirad, Akira Kamiya

研究成果: Article査読

8 被引用数 (Scopus)

抄録

By designing a new epi-illumination system with dual slit laser beams, we have developed a fluorescence microscope for tomographic observations of in vivo microcirculation. Two beams of an Argon ion laser were converted into thin slit beams of thickness 28 to 60 μm by optical lenses and were illuminated onto tissue to intersect at the focal plane of the objective. The fluorescent light emitted from a FITC tracer by this cross-illumination was picked up by the microscope through a filter, with no “out of focus” noise from the tracer above and below the crossing zone. The results of in vitro experiments by using glass micropipettes (20-80 μm i.d.) filled with FITC-dextran and skeletal muscle tissue certified uniform tracer excitation in the crossing zone and the augmentation of the beam thickness due to light scattering in the tissue to be 1.7 and 2.5 times at depths of 100 and 200 μm from the surface, respectively. In vivo tests in the rabbit tenuissimus muscle revealed that this system can realize microvasculature tomography, with adequate vertical zone selectivity and spatial resolution to reconstruct its three-dimensional mapping, and long-term monitoring of tracer leakage from a single capillary to the surrounding tissue, with sufficient quantitative reliability to determine the capillary permeability and its heterogeneity along the channel.

本文言語English
ページ(範囲)300-314
ページ数15
ジャーナルMicrovascular Research
49
3
DOI
出版ステータスPublished - 1995 5月
外部発表はい

ASJC Scopus subject areas

  • 生化学
  • 循環器および心血管医学
  • 細胞生物学

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