Single-molecule kinetics in living cells

TL;DR

This paper reviews how single-molecule microscopy techniques enable detailed study of biomolecular kinetics within living cells, highlighting their advantages and addressing associated challenges.

Contribution

It provides a comprehensive overview of single-molecule methods applied to live-cell kinetics, emphasizing recent technological advances and solutions to methodological challenges.

Findings

Single-molecule techniques reveal intramolecular and intermolecular kinetics in vivo.

These methods offer insights not accessible through ensemble measurements.

Challenges include localization precision and tracking in complex cellular environments.

Abstract

In the past decades, advances in microscopy have made it possible to study the dynamics of individual biomolecules in vitro and resolve intramolecular kinetics that would otherwise be hidden in ensemble averages. More recently, single-molecule methods have been used to image, localize and track individually labeled macromolecules in the cytoplasm of living cells, allowing investigations of intermolecular kinetics under physiologically relevant conditions. In this review, we illuminate the particular advantages of single-molecule techniques when studying kinetics in living cells and discuss solutions to specific challenges associated with these methods.