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Anal Biochem. 2011 Aug 15;415(2):182-9. doi: 10.1016/j.ab.2011.03.039. Epub 2011 Apr 6.

Video rate bioluminescence imaging of secretory proteins in living cells: localization, secretory frequency, and quantification.

Author information

1
Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya 464-8650, Japan.

Abstract

We have developed a method of video rate bioluminescence imaging to investigate protein secretion from a single mammalian cell and analyzed the localization, secretory frequency, and quantification of secreted protein. By detecting the luminescence signals from the Gaussia luciferase (GLase) reaction using a high-speed electron-multiplying charge-coupled device (EM-CCD) camera, video rate imaging was performed with a time resolution within 500 ms/image over 30 min in living cells. As a model study, we applied the method to visualize the glucose-stimulated insulin secretion from clustered pancreatic MIN6 β cells using the fused protein of GLase with preproinsulin. High-quality video images clearly showed that the glucose-stimulated insulin secretion from the clustered MIN6 β cells oscillated within a period of a few minutes over 10 min. In addition, the glibenclamide-induced insulin secretion from the clustered MIN6 β cells was visualized, suggesting that bioluminescence video rate imaging is a useful method for validating drug action in living cells.

PMID:
21477579
DOI:
10.1016/j.ab.2011.03.039
[Indexed for MEDLINE]

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