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

Result code in 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>

Result on the web

<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>

Result language

angličtina

Original language name

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

Original language description

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

Czech name

—

Czech description

—

Type

D - Article in proceedings

CEP classification

—

OECD FORD branch

10306 - Optics (including laser optics and quantum optics)

Project

<a href="/en/project/LL1602" target="_blank" >LL1602: Optical imaging of single protein dynamics</a><br>

Continuities

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

Publication year

2018

Confidentiality

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

Article name in the collection

NANOIMAGING AND NANOSPECTROSCOPY VI

ISBN

978-1-5106-2024-7

ISSN

0277-786X

e-ISSN

1996-756X

Number of pages

8

Pages from-to

—

Publisher name

SPIE

Place of publication

San Diego

Event location

San Diego

Event date

Aug 19, 2018

Type of event by nationality

WRD - Celosvětová akce

UT code for WoS article

000451508900009