Issue 6, 2023

In situ silver nanoparticle development for molecular-specific biological imaging via highly accessible microscopies

Abstract

In biological studies and diagnoses, brightfield (BF), fluorescence, and electron microscopy (EM) are used to image biomolecules inside cells. When compared, their relative advantages and disadvantages are obvious. BF microscopy is the most accessible of the three, but its resolution is limited to a few microns. EM provides a nanoscale resolution, but sample preparation is time-consuming. In this study, we present a new imaging technique, which we termed decoration microscopy (DecoM), and quantitative investigations to address the aforementioned issues in EM and BF microscopy. For molecular-specific EM imaging, DecoM labels proteins inside cells using antibodies bearing 1.4 nm gold nanoparticles (AuNPs) and grows silver layers on the AuNPs' surfaces. The cells are then dried without buffer exchange and imaged using scanning electron microscopy (SEM). Structures labeled with silver-grown AuNPs are clearly visible on SEM, even they are covered with lipid membranes. Using stochastic optical reconstruction microscopy, we show that the drying process causes negligible distortion of structures and that less structural deformation could be achieved through simple buffer exchange to hexamethyldisilazane. Using DecoM, we visualize the nanoscale alterations in microtubules by microtubule-severing proteins that cannot be observed with diffraction-limited fluorescence microscopy. We then combine DecoM with expansion microscopy to enable sub-micron resolution BF microscopy imaging. We first show that silver-grown AuNPs strongly absorb white light, and the structures labeled with them are clearly visible on BF microscopy. We then show that the application of AuNPs and silver development must follow expansion to visualize the labeled proteins clearly with sub-micron resolution.

Graphical abstract: In situ silver nanoparticle development for molecular-specific biological imaging via highly accessible microscopies

Supplementary files

Article information

Article type
Paper
Submitted
13 Jūl. 2022
Accepted
19 Dec. 2022
First published
21 Dec. 2022
This article is Open Access
Creative Commons BY-NC license

Nanoscale Adv., 2023,5, 1636-1650

In situ silver nanoparticle development for molecular-specific biological imaging via highly accessible microscopies

D. Song, C. W. Song, J. Chung, E. Jang, H. Kim, Y. Hur, E. Hur, D. Kim and J. Chang, Nanoscale Adv., 2023, 5, 1636 DOI: 10.1039/D2NA00449F

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