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Pulsed interleaved excitation-based line-scanning spatial correlation spectroscopy (PIE-lsSCS)

Gao, X.; Gao, P.; Prunsche, B.; Nienhaus, K.; Nienhaus, G. U.

Abstract:
We report pulsed interleaved excitation (PIE) based line-scanning spatial correlation spectroscopy (PIElsSCS), a quantitative fluorescence microscopy method for the study of dynamics in free-standing lipid bilayer membranes. Using a confocal microscope, we scan multiple lines perpendicularly through the membrane, each one laterally displaced from the previous one by several ten nanometers. Scanning through the membrane enables us to eliminate intensity fluctuations due to membrane displacements with respect to the observation volume. The diffusion of fluorescent molecules within the membrane is quantified by spatial correlation analysis, based on the fixed lag times between successive line scans. PIE affords dual-color excitation within a single line scan and avoids channel crosstalk. PIE-lsSCS data are acquired from a larger membrane region so that sampling is more efficient. Moreover, the local photon flux is reduced compared with single-point experiments, resulting in a smaller fraction of photobleached molecules for identical exposure times. This is helpful for precise measurements on live cells and tissues. We have evaluated th ... mehr

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Verlagsausgabe §
DOI: 10.5445/IR/1000087861
Veröffentlicht am 26.11.2018
Seitenaufrufe: 32
seit 27.11.2018
Downloads: 23
seit 30.11.2018
Zugehörige Institution(en) am KIT Institut für Angewandte Physik (APH)
Institut für Toxikologie und Genetik (ITG)
Institut für Nanotechnologie (INT)
Publikationstyp Zeitschriftenaufsatz
Jahr 2018
Sprache Englisch
Identifikator ISSN: 2045-2322
urn:nbn:de:swb:90-878611
KITopen-ID: 1000087861
HGF-Programm 43.23.01 (POF III, LK 01)
Erschienen in Scientific reports
Band 8
Heft 1
Seiten Art. Nr.: 16722
Bemerkung zur Veröffentlichung Gefördert durch den KIT-Publikationsfonds
Nachgewiesen in Scopus
Web of Science
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