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FLUORESCENCE SPECTROSCOPY AND MICROSCOPY: METHODS AND PROTOCOLS
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SYSNO ASEP 0506914 Druh ASEP M - Kapitola v monografii Zařazení RIV C - Kapitola v knize Název Z-Scan Fluorescence Correlation Spectroscopy as a Tool for Diffusion Measurements in Planar Lipid Membranes Tvůrce(i) Steinberger, T. (CZ)
Macháň, Radek (UFCH-W)
Hof, Martin (UFCH-W) RID, ORCIDZdroj.dok. FLUORESCENCE SPECTROSCOPY AND MICROSCOPY: METHODS AND PROTOCOLS, Methods and Protocols. - Totowa : HUMANA PRESS INC, 2014 / Engelborghs Y. ; Visser A.J.W.G. - ISSN 1064-3745 - ISBN 978-1-62703-648-1 Rozsah stran roč. 1076 (2014), s. 617-634 Edice Methods in Molecular Biology Poč.str. 18 s. Poč.str.knihy 817 Forma vydání Tištěná - P Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova lifetime correlation spectroscopy ; optical saturation ; continuous-wave ; bilayers ; coefficients ; accuracy ; systems ; model ; mica Vědní obor RIV CF - Fyzikální chemie a teoretická chemie Obor OECD Physical chemistry CEP GAP208/10/0376 GA ČR - Grantová agentura ČR Institucionální podpora UFCH-W - RVO:61388955 UT WOS 000328374800029 EID SCOPUS 84934443471 DOI 10.1007/978-1-62703-649-8_28 Anotace Studies of lateral diffusion are used for the characterization of the dynamics of biological membranes. One of the techniques that can be used for this purpose is fluorescence correlation spectroscopy (FCS), which belongs to the single-molecule techniques. Unfortunately, FCS measurements, when performed in planar lipid systems, are associated with a few sources of inaccuracy in the determination of the lateral diffusion coefficient. The main problems are related to the imperfect positioning of the laser focus relative to the plane of the sample. Another source of inaccuracy is the requirement for external calibration of the detection volume size. This protocol introduces a calibration-free method called Z-scan fluorescence correlation spectroscopy (Z-scan FCS), which is based on the determination of the diffusion time and particle number in steps along the optical (z-) axis by sequential FCS measurements. Z-scan FCS could be employed for diffusion measurements in planar membrane model systems-supported phospholipid bilayers (SPBs) and giant unilamellar vesicles (GUVs) and also in biological membranes. A result from measurements in SPBs is also presented in the protocol as a principle example of the Z-scan technique. Pracoviště Ústav fyzikální chemie J.Heyrovského Kontakt Michaela Knapová, michaela.knapova@jh-inst.cas.cz, Tel.: 266 053 196 Rok sběru 2020
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