Progress in quantitative single-molecule localization microscopy.
Histochem Cell Biol
; 142(1): 5-17, 2014 Jul.
Article
in En
| MEDLINE
| ID: mdl-24748502
ABSTRACT
With the advent of single-molecule localization microscopy (SMLM) techniques, intracellular proteins can be imaged at unprecedented resolution with high specificity and contrast. These techniques can lead to a better understanding of cell functioning, as they allow, among other applications, counting the number of molecules of a protein specie in a single cell, studying the heterogeneity in protein spatial organization, and probing the spatial interactions between different protein species. However, the use of these techniques for accurate quantitative measurements requires corrections for multiple inherent sources of error, including overcounting due to multiple localizations of a single fluorophore (i.e., photoblinking), undercounting caused by incomplete photoconversion, uncertainty in the localization of single molecules, sample drift during the long imaging time, and inaccurate image registration in the case of dual-color imaging. In this paper, we review recent efforts that address some of these sources of error in quantitative SMLM and give examples in the context of photoactivated localization microscopy (PALM).
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Proteins
/
Molecular Imaging
/
Microscopy, Fluorescence
Language:
En
Journal:
Histochem Cell Biol
Year:
2014
Document type:
Article