Development of a novel cryogenic microscope with numerical aperture of 0.9 and its application to photosynthesis research

Yutaka Shibata, Wataru Katoh, Tomofumi Chiba, Keisuke Namie, Norikazu Ohnishi, Jun Minagawa, Hanayo Nakanishi, Takumi Noguchi, Hiroshi Fukumura

研究成果: Article査読

9 被引用数 (Scopus)

抄録

A novel cryogenic optical-microscope system was developed in which the objective lens is set inside of the cryostat adiabatic vacuum space. Being isolated from the sample when it was cooled, the objective lens was maintained at room temperature during the cryogenic measurement. Therefore, the authors were able to use a color-aberration corrected objective lens with a numerical aperture of 0.9. The lens is equipped with an air vent for compatibility to the vacuum. The theoretically expected spatial resolutions of 0.39 μm along the lateral direction and 1.3 μm along the axial direction were achieved by the developed system. The system was applied to the observations of non-uniform distributions of the photosystems in the cells of a green alga, Chlamydomonas reinhardtii, at 94 K. Gaussian decomposition analysis of the fluorescence spectra at all the pixels clearly demonstrated a non-uniform distribution of the two photosystems, as reflected in the variable ratios of the fluorescence intensities assigned to photosystem II and to those assigned to photosystem I. The system was also applied to the fluorescence spectroscopy of single isolated photosystem I complexes at 90 K. The fluorescence, assigned to be emitted from a single photosystem I trimer, showed an intermittent fluctuation called blinking, which is typical for a fluorescence signal from a single molecule. The vibronic fluorescence bands at around 790 nm were observed for single photosystem I trimers, suggesting that the color aberration is not serious up to the 800 nm spectral region.

本文言語English
ページ(範囲)880-887
ページ数8
ジャーナルBiochimica et Biophysica Acta - Bioenergetics
1837
6
DOI
出版ステータスPublished - 2014 6

ASJC Scopus subject areas

  • 生物理学
  • 生化学
  • 細胞生物学

フィンガープリント

「Development of a novel cryogenic microscope with numerical aperture of 0.9 and its application to photosynthesis research」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル