Visualization and intracellular dynamic tracking of exosomes and exosomal miRNAs using single molecule localization microscopy

Nanoscale
Chen ChenYiping Cui

Abstract

Exosomes are small membrane vesicles secreted by a wide variety of cells. Studies have demonstrated that exosomal miRNAs can influence the biological processes of recipient cells. Therefore, direct imaging and tracking of exosomal miRNAs in living recipient cells are essential for exosome functional analysis. However, the moderate spatial resolution of conventional fluorescence microscopy limits the precise imaging and tracking of exosomes considering their relatively small size (<100 nm). Here, we took advantage of single molecule localization microscopy (SMLM) to realize the visualization and dynamic tracking of exosomes and exosomal miRNAs in living cells. First, we demonstrated simultaneous SMLM imaging of HeLa-derived exosomes and two kinds of exosomal miRNAs (mir-21 and mir-31) and tracked their movement after cellular uptake. The motion of exosomes within the intercellular filamentous structures was observed successfully. Moreover, dual-color SMLM based dynamic tracking revealed that as one kind of natural carrier of cellular cargoes, exosomes can encapsulate mir-21 inside to prevent enzyme degradation during transfer and then release them into recipient cells. These findings can provide new insights into the pathway of ...Continue Reading

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Citations

Nov 7, 2019·Chemical Communications : Chem Comm·Fang HeBang-Ce Ye
Jul 2, 2020·International Journal of Nanomedicine·Yan WangQi Li
Nov 2, 2018·International Journal of Molecular Sciences·Abdelnaby KhalyfaDavid Gozal
Feb 1, 2020·Journal of Extracellular Vesicles·Maria S PanagopoulouChristopher D Gregory
Jul 2, 2019·Journal of Extracellular Vesicles·Yongwei XiaoTianyu Zhong
Sep 27, 2018·Proceedings of the National Academy of Sciences of the United States of America·Yuting YangHui Yu
Mar 21, 2019·International Journal of Molecular Sciences·Sabrina RoyBob S Carter
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Apr 25, 2021·Cell Communication and Signaling : CCS·Sonam GurungJulien Baruteau
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Aug 8, 2021·Journal of Clinical Medicine·Anne-Tess C JolinkRick Kapur
Apr 14, 2020·ACS Nano·Bhagyashree S JoshiInge S Zuhorn

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Methods Mentioned

BETA
fluorescence microscopy
imaging techniques
transmission
atomic force microscopy
AFM
fluorescence imaging
cleavage assay
Precipitation
transmission electron microscopy

Software Mentioned

Adobe After Effects CC
Zeiss Elyra

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