Quantifying F-actin patches in single melanoma cells using total-internal reflection fluorescence microscopy.

ABSTRACT: Total-internal reflection fluorescence (TIRF) microscope is a unique technique for selective excitation of only those fluorophore molecules in a cellular environment, which are located at the sub-diffraction axial distance of a cell's contact-area. Despite this prominent feature of the TIRF microscope, making quantitative use of this technique has been a challenge, since the excitation intensity strongly depends on the axial position of a fluorophore molecule. Here, we present an easy-implemented data analysis method to quantitatively characterize the fluorescent signal, without considering the intensity-value. We use F-actin patches in single-melanoma cells as an example and define two quantities of elongation and surface density for F-actin patches at the contact-area of a melanoma cell.

SUBMITTER: Sheykhi E 

PROVIDER: S-EPMC9678867 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature