Trends in Single-Molecule Total Internal Reflection Fluorescence Imaging and Their Biological Applications with Lab-on-a-Chip Technology

Author:

Colson Louis1ORCID,Kwon Youngeun2,Nam Soobin2,Bhandari Avinashi1,Maya Nolberto Martinez1,Lu Ying1,Cho Yongmin2ORCID

Affiliation:

1. Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA

2. Department of Chemical Engineering, Myongji University, Yongin 17058, Republic of Korea

Abstract

Single-molecule imaging technologies, especially those based on fluorescence, have been developed to probe both the equilibrium and dynamic properties of biomolecules at the single-molecular and quantitative levels. In this review, we provide an overview of the state-of-the-art advancements in single-molecule fluorescence imaging techniques. We systematically explore the advanced implementations of in vitro single-molecule imaging techniques using total internal reflection fluorescence (TIRF) microscopy, which is widely accessible. This includes discussions on sample preparation, passivation techniques, data collection and analysis, and biological applications. Furthermore, we delve into the compatibility of microfluidic technology for single-molecule fluorescence imaging, highlighting its potential benefits and challenges. Finally, we summarize the current challenges and prospects of fluorescence-based single-molecule imaging techniques, paving the way for further advancements in this rapidly evolving field.

Funder

Myongji University

Publisher

MDPI AG

Subject

Electrical and Electronic Engineering,Biochemistry,Instrumentation,Atomic and Molecular Physics, and Optics,Analytical Chemistry

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