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Unravelling the Mystery inside Cells by Using Single-Molecule Fluorescence Imaging

Live-cell imaging is a powerful technique to study the dynamics and mechanics of various biological molecules like proteins, organelles, DNA, and RNA. With the rapid evolution of optical microscopy, our understanding of how these molecules are implicated in the cells’ most critical physiological rol...

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Autores principales: Zalejski, Julian, Sun, Jiachen, Sharma, Ashutosh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10532472/
https://www.ncbi.nlm.nih.gov/pubmed/37754956
http://dx.doi.org/10.3390/jimaging9090192
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author Zalejski, Julian
Sun, Jiachen
Sharma, Ashutosh
author_facet Zalejski, Julian
Sun, Jiachen
Sharma, Ashutosh
author_sort Zalejski, Julian
collection PubMed
description Live-cell imaging is a powerful technique to study the dynamics and mechanics of various biological molecules like proteins, organelles, DNA, and RNA. With the rapid evolution of optical microscopy, our understanding of how these molecules are implicated in the cells’ most critical physiological roles deepens. In this review, we focus on how spatiotemporal nanoscale live-cell imaging at the single molecule level allows for profound contributions towards new discoveries in life science. This review will start by summarizing how single-molecule tracking has been used to analyze membrane dynamics, receptor–ligand interactions, protein–protein interactions, inner- and extra-cellular transport, gene expression/transcription, and whole organelle tracking. We then move on to how current authors are trying to improve single-molecule tracking and overcome current limitations by offering new ways of labeling proteins of interest, multi-channel/color detection, improvements in time-lapse imaging, and new methods and programs to analyze the colocalization and movement of targets. We later discuss how single-molecule tracking can be a beneficial tool used for medical diagnosis. Finally, we wrap up with the limitations and future perspectives of single-molecule tracking and total internal reflection microscopy.
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spelling pubmed-105324722023-09-28 Unravelling the Mystery inside Cells by Using Single-Molecule Fluorescence Imaging Zalejski, Julian Sun, Jiachen Sharma, Ashutosh J Imaging Review Live-cell imaging is a powerful technique to study the dynamics and mechanics of various biological molecules like proteins, organelles, DNA, and RNA. With the rapid evolution of optical microscopy, our understanding of how these molecules are implicated in the cells’ most critical physiological roles deepens. In this review, we focus on how spatiotemporal nanoscale live-cell imaging at the single molecule level allows for profound contributions towards new discoveries in life science. This review will start by summarizing how single-molecule tracking has been used to analyze membrane dynamics, receptor–ligand interactions, protein–protein interactions, inner- and extra-cellular transport, gene expression/transcription, and whole organelle tracking. We then move on to how current authors are trying to improve single-molecule tracking and overcome current limitations by offering new ways of labeling proteins of interest, multi-channel/color detection, improvements in time-lapse imaging, and new methods and programs to analyze the colocalization and movement of targets. We later discuss how single-molecule tracking can be a beneficial tool used for medical diagnosis. Finally, we wrap up with the limitations and future perspectives of single-molecule tracking and total internal reflection microscopy. MDPI 2023-09-19 /pmc/articles/PMC10532472/ /pubmed/37754956 http://dx.doi.org/10.3390/jimaging9090192 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Review
Zalejski, Julian
Sun, Jiachen
Sharma, Ashutosh
Unravelling the Mystery inside Cells by Using Single-Molecule Fluorescence Imaging
title Unravelling the Mystery inside Cells by Using Single-Molecule Fluorescence Imaging
title_full Unravelling the Mystery inside Cells by Using Single-Molecule Fluorescence Imaging
title_fullStr Unravelling the Mystery inside Cells by Using Single-Molecule Fluorescence Imaging
title_full_unstemmed Unravelling the Mystery inside Cells by Using Single-Molecule Fluorescence Imaging
title_short Unravelling the Mystery inside Cells by Using Single-Molecule Fluorescence Imaging
title_sort unravelling the mystery inside cells by using single-molecule fluorescence imaging
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10532472/
https://www.ncbi.nlm.nih.gov/pubmed/37754956
http://dx.doi.org/10.3390/jimaging9090192
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