Combining PALM and SOFI for quantitative imaging of focal adhesions in living cells

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68407700%3A21230%2F17%3A00349482" target="_blank" >RIV/68407700:21230/17:00349482 - isvavai.cz</a>
Result on the web
<a href="https://doi.org/10.1117/12.2252865" target="_blank" >https://doi.org/10.1117/12.2252865</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1117/12.2252865" target="_blank" >10.1117/12.2252865</a>

Result language
angličtina
Original language name
Combining PALM and SOFI for quantitative imaging of focal adhesions in living cells
Original language description
Focal adhesions are complicated assemblies of hundreds of proteins that allow cells to sense their extracellular matrix and adhere to it. Although most focal adhesion proteins have been identified, their spatial organization in living cells remains challenging to observe. Photo-activated localization microscopy (PALM) is an interesting technique for this purpose, especially since it allows estimation of molecular parameters such as the number of fluorophores. However, focal adhesions are dynamic entities, requiring a temporal resolution below one minute, which is difficult to achieve with PALM. In order to address this problem, we merged PALM with super-resolution optical fluctuation imaging (SOFI) by applying both techniques to the same data. Since SOFI tolerates an overlap of single molecule images, it can improve the temporal resolution compared to PALM. Moreover, an adaptation called balanced SOFI (bSOFI) allows estimation of molecular parameters, such as the fluorophore density. We therefore performed simulations in order to assess PALM and SOFI for quantitative imaging of dynamic structures. We demonstrated the potential of our PALM-SOFI concept as a quantitative imaging framework by investigating moving focal adhesions in living cells.
Czech name
—
Czech description
—

Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Article name in the collection
Single Molecule Spectroscopy and Superresolution Imaging X
ISBN
978-1-5106-0583-1
ISSN
0277-786X
e-ISSN
2410-9045
Number of pages
7
Pages from-to
—
Publisher name
SPIE
Place of publication
Bellingham (stát Washington)
Event location
San Francisco
Event date
Jan 28, 2017
Type of event by nationality
WRD - Celosvětová akce
UT code for WoS article
000406422600002

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