Slit-scanning microscope with a high-NA objective lens for analysis of synaptic function

Takashi Sakurai; Yoshihiko Wakazono; Seiji Yamamoto M.D.; Susumu Terakawa M.D.

doi:10.1117/12.529115

1 July 2004 Slit-scanning microscope with a high-NA objective lens for analysis of synaptic function

Takashi Sakurai, Yoshihiko Wakazono, Seiji Yamamoto M.D., Susumu Terakawa M.D.

Proceedings Volume 5322, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II; (2004) https://doi.org/10.1117/12.529115
Event: Biomedical Optics 2004, 2004, San Jose, CA, United States

Abstract

By employing the total internal reflection fluorescence (TIRF) microscope with an ultra high NA (1.65) objective lens, we demonstrated detailed dynamics of exocytosis in various types of secretory vesicles. However, the TIRF microscopy could be applied to observations only on the plasma membrane and its immediate vicinity. To observe the vesicles in the deeper region of cytoplasm, we modified the TIRF optics to project a slit beam thinner than 1 μm in width to the cell. The slit beam illumination spotted single secretory vesicles inside the cell better and their movement and exocytosis easier. By scanning the slit beam, a fluorescence microscopy was possible at a high signal-to-noise ratio useful for measurement and analysis of single exocytosis in neurons and endocrine cells.

Citation Download Citation

Takashi Sakurai, Yoshihiko Wakazono, Seiji Yamamoto M.D., and Susumu Terakawa M.D. "Slit-scanning microscope with a high-NA objective lens for analysis of synaptic function", Proc. SPIE 5322, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II, (1 July 2004); https://doi.org/10.1117/12.529115

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KEYWORDS

Luminescence

Microscopes

Microscopy

Neurons

Objectives

Glasses

Molecules

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