Interferometric scattering (iSCAT) microscopy for high fidelity tracking at microseconds timescales

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985882%3A_____%2F18%3A00500648" target="_blank" >RIV/67985882:_____/18:00500648 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1117/12.2321086" target="_blank" >http://dx.doi.org/10.1117/12.2321086</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1117/12.2321086" target="_blank" >10.1117/12.2321086</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Interferometric scattering (iSCAT) microscopy for high fidelity tracking at microseconds timescales

Popis výsledku v původním jazyce

Novel methods aiming at understanding complex biophysical processes allow revealing the dynamics and behaviour in extreme detail down to a single protein. Developments of fluorescence-based super-resolution microscopy and nanoscopic tracking techniques helped to reach a spatial resolution in length scales below 10 nm. These advances rely on the efficient collection of fluorescence at single-molecule levels. However, complex photophysics and saturation of fluorescent labels limit the temporal resolution to milliseconds timescales. To overcome the spatiotemporal limitations of fluorescent-based techniques we are employing interferometric scattering microscopy (iSCAT). iSCAT is an optical microscopy technique which allows for the detection and localization of extremely low scattering signals. It is based on interference of light scattered on the particle with a reference wave, e.g. light partially reflected at a glass coverslip. The sensitivity of iSCAT was previously proven in detection experiments with small nanoparticles as well as unlabelled single proteins. Here, we show that scattering labels can be imaged and localized with a nanometer precision and a few microseconds temporal resolution. We investigate the limits of fast tracking of scattering labels and identify pitfalls of high-speed collection for which the tracking fidelity drops rapidly due to fluctuations in the label position

Název v anglickém jazyce

Interferometric scattering (iSCAT) microscopy for high fidelity tracking at microseconds timescales

Popis výsledku anglicky

Novel methods aiming at understanding complex biophysical processes allow revealing the dynamics and behaviour in extreme detail down to a single protein. Developments of fluorescence-based super-resolution microscopy and nanoscopic tracking techniques helped to reach a spatial resolution in length scales below 10 nm. These advances rely on the efficient collection of fluorescence at single-molecule levels. However, complex photophysics and saturation of fluorescent labels limit the temporal resolution to milliseconds timescales. To overcome the spatiotemporal limitations of fluorescent-based techniques we are employing interferometric scattering microscopy (iSCAT). iSCAT is an optical microscopy technique which allows for the detection and localization of extremely low scattering signals. It is based on interference of light scattered on the particle with a reference wave, e.g. light partially reflected at a glass coverslip. The sensitivity of iSCAT was previously proven in detection experiments with small nanoparticles as well as unlabelled single proteins. Here, we show that scattering labels can be imaged and localized with a nanometer precision and a few microseconds temporal resolution. We investigate the limits of fast tracking of scattering labels and identify pitfalls of high-speed collection for which the tracking fidelity drops rapidly due to fluctuations in the label position

Druh

D - Stať ve sborníku

CEP obor

—

OECD FORD obor

10306 - Optics (including laser optics and quantum optics)

Projekt

<a href="/cs/project/LL1602" target="_blank" >LL1602: Optické zobrazování dynamiky jednotlivých proteinů</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2018

Kód důvěrnosti údajů

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

Název statě ve sborníku

NANOIMAGING AND NANOSPECTROSCOPY VI

ISBN

978-1-5106-2024-7

ISSN

0277-786X

e-ISSN

1996-756X

Počet stran výsledku

8

Strana od-do

—

Název nakladatele

SPIE

Místo vydání

San Diego

Místo konání akce

San Diego

Datum konání akce

19. 8. 2018

Typ akce podle státní příslušnosti

WRD - Celosvětová akce

Kód UT WoS článku

000451508900009