Fluorescence correlation spectroscopy diffusion laws in the presence of moving nanodomains

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388955%3A_____%2F16%3A00457556" target="_blank" >RIV/61388955:_____/16:00457556 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1088/0022-3727/49/11/114002" target="_blank" >http://dx.doi.org/10.1088/0022-3727/49/11/114002</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1088/0022-3727/49/11/114002" target="_blank" >10.1088/0022-3727/49/11/114002</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Fluorescence correlation spectroscopy diffusion laws in the presence of moving nanodomains

Popis výsledku v původním jazyce

It has been shown by means of simulations that spot variation fluorescence correlation/nspectroscopy (sv-FCS) can be used for the identification and, to some extent, also/ncharacterization of immobile lipid nanodomains in model as well as cellular plasma/nmembranes. However, in these simulations, the nanodomains were assumed to be stationary,/nwhereas they actually tend to move like the surrounding lipids. In the present study, we/ninvestigated how such domain movement influences the diffusion time/spot-size dependence/nobserved in FCS experiments, usually referred to as diffusion law’ analysis. We show/nthat domain movement might mask the effects of the anomalous’ diffusion characteristics/nof membrane lipids or proteins predicted for stationary domains, making it difficult to/nidentify such moving nanodomains by sv-FCS. More specifically, our simulations indicate/nthat (i) for domains moving up to a factor of 2.25 slower than the surrounding lipids, such/nimpeded diffusion cannot be observed and the diffusion behaviour of the proteins or lipids is/nindistinguishable from that of freely diffusing molecules, i.e. nanodomains are not detected;/n(ii) impeded protein/lipid diffusion behaviour can be observed in experiments where the radii/nof the detection volume are similar in size to the domain radii, the domain diffusion is about/n10 times slower than that of the lipids, and the probes show a high affinity to the domains;/nand (iii) presence of nanodomains can only be reliably detected by diffraction limited sv-FCS/nwhen the domains move very slowly (about 200 times slower than the lipid diffusion). As/nnanodomains are expected to be in the range of tens of nanometres and most probes show/nlow affinities to such domains, sv-FCS is limited to stationary domains and/or STED-FCS./nHowever, even for that latter technique, diffusing domains smaller than 50 nm in radius are/nhardly detectable by FCS diffusion time/spot-size dependencies.

Název v anglickém jazyce

Fluorescence correlation spectroscopy diffusion laws in the presence of moving nanodomains

Popis výsledku anglicky

It has been shown by means of simulations that spot variation fluorescence correlation/nspectroscopy (sv-FCS) can be used for the identification and, to some extent, also/ncharacterization of immobile lipid nanodomains in model as well as cellular plasma/nmembranes. However, in these simulations, the nanodomains were assumed to be stationary,/nwhereas they actually tend to move like the surrounding lipids. In the present study, we/ninvestigated how such domain movement influences the diffusion time/spot-size dependence/nobserved in FCS experiments, usually referred to as diffusion law’ analysis. We show/nthat domain movement might mask the effects of the anomalous’ diffusion characteristics/nof membrane lipids or proteins predicted for stationary domains, making it difficult to/nidentify such moving nanodomains by sv-FCS. More specifically, our simulations indicate/nthat (i) for domains moving up to a factor of 2.25 slower than the surrounding lipids, such/nimpeded diffusion cannot be observed and the diffusion behaviour of the proteins or lipids is/nindistinguishable from that of freely diffusing molecules, i.e. nanodomains are not detected;/n(ii) impeded protein/lipid diffusion behaviour can be observed in experiments where the radii/nof the detection volume are similar in size to the domain radii, the domain diffusion is about/n10 times slower than that of the lipids, and the probes show a high affinity to the domains;/nand (iii) presence of nanodomains can only be reliably detected by diffraction limited sv-FCS/nwhen the domains move very slowly (about 200 times slower than the lipid diffusion). As/nnanodomains are expected to be in the range of tens of nanometres and most probes show/nlow affinities to such domains, sv-FCS is limited to stationary domains and/or STED-FCS./nHowever, even for that latter technique, diffusing domains smaller than 50 nm in radius are/nhardly detectable by FCS diffusion time/spot-size dependencies.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CF - Fyzikální chemie a teoretická chemie

OECD FORD obor

—

Projekt

<a href="/cs/project/GC14-03141J" target="_blank" >GC14-03141J: Studium vztahu struktury a funkce FGF2 oligomerů tvořících membránové póry technikami zaměřenými na sledování jedné molekuly</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2016

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Physics D-Applied Physics

ISSN

0022-3727

e-ISSN

—

Svazek periodika

49

Číslo periodika v rámci svazku

11

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

11

Strana od-do

—

Kód UT WoS článku

000371007100003

EID výsledku v databázi Scopus

2-s2.0-84960096929