A method for extraction of exosomes from breast tumour cells and characterisation by transmission electron microscopy

Author:

Liu Zhiang1ORCID,Xue Haowen2,Chen Qi234,Yang Ge2

Affiliation:

1. TEM Laboratory Experimental Teaching and Equipment Management Center Qufu Normal University Qufu Shandong People's Republic of China

2. College of Life Sciences Qufu Normal University Qufu Shandong People's Republic of China

3. College of Agriculture Fujian Agriculture and Forestry University Fuzhou People's Republic of China

4. Basic Forestry and Proteomics Research Center Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou People's Republic of China

Abstract

AbstractExosomes can not only be used as markers of tumour metastasis but also be used for noninvasive diagnosis of clinical diseases, which holds significant medical research value. Observing the morphology and distribution of exosomes using transmission electron microscopy (TEM) is highly significant. In this study, we use breast tumour cell (MDA‐MB‐231) exosomes as a representative sample and focus on the extraction and purification of exosomes, as well as the investigation of optimal conditions for the observation of exosomes using TEM. Through comparative tests, we obtained the optimal dilution concentration and staining method for TEM of exosomes, the best dilution concentration is 100 times, the best negative staining time is 1.5 min. Western blotting and scanning electron microscopy (SEM) confirmed the presence of exosome. The particle size experiment shows that the size of exosomes is mainly distributed between 60 nm and 150 nm. This method provides a reference for TEM sample preparation of subcellular structures and small molecular biomaterials other than exosomes.

Funder

Natural Science Foundation of Shandong Province

Publisher

Wiley

Subject

Histology,Pathology and Forensic Medicine

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