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  Tracking single secretory granules in live chromaffin cells by evanescent-field fluorescence microscopy

Steyer, J. A., & Almers, W. (1999). Tracking single secretory granules in live chromaffin cells by evanescent-field fluorescence microscopy. Biophysical Journal, 76(4), 2262-2271. doi:10.1016/S0006-3495(99)77382-0.

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Item Permalink: https://hdl.handle.net/11858/00-001M-0000-0024-9273-1 Version Permalink: https://hdl.handle.net/11858/00-001M-0000-002D-A9D4-9
Genre: Journal Article

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BiophysJ_76_1999_2262.pdf (Any fulltext), 299KB
 
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 Creators:
Steyer, Jürgen A.1, Author           
Almers, Wolfhard1, Author           
Affiliations:
1Department of Molecular Cell Research, Max Planck Institute for Medical Research, Max Planck Society, ou_1497703              

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 Abstract: We have observed secretory granules beneath the plasma membrane of chromaffin cells. Using evanescent-field excitation by epiillumination, we have illuminated a thin layer of cytosol where cells adhere to glass coverslips. Up to 600 frames could be recorded at diffraction-limited resolution without appreciable photodynamic damage. We localized single granules with an uncertainty of ∼30 nm and tracked their motion in three dimensions. Granules in resting cells wander randomly as if imprisoned in a cage that leaves ∼70 nm space around a granule. The “cage” itself moves only slowly (D = 2 × 10−12 cm2/s). Rarely do granules arrive at or depart from the plasma membrane of resting cells. Stimulation increases lateral motion only slightly. After the plasma membrane has been depleted of granules by exocytosis, fresh granules can be seen to approach it at an angle. The method will be useful for exploring the molecular steps preceding exocytosis at the level of single granules.

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Language(s): eng - English
 Dates: 1998-10-151998-08-171998-10-151999-04
 Publication Status: Issued
 Pages: 10
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 666531
DOI: 10.1016/S0006-3495(99)77382-0
URI: http://www.ncbi.nlm.nih.gov/pubmed/10096921
URI: http://www.sciencedirect.com/science/article/pii/S0006349599773820
Other: 4478
 Degree: -

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Title: Biophysical Journal
  Other : Biophys. J.
Source Genre: Journal
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Publ. Info: Cambridge, Mass. : Cell Press
Pages: - Volume / Issue: 76 (4) Sequence Number: - Start / End Page: 2262 - 2271 Identifier: Other: 0006-3495
CoNE: https://pure.mpg.de/cone/journals/resource/954925385117