Precision and accuracy of receptor quantification on synthetic and biological surfaces using DNA-PAINT

Roger Riera Brillas, Emmanouil Archontakis, Glenn A.O. Cremers, Tom F.A. de Greef, Peter Zijlstra (Corresponding author), Lorenzo Albertazzi (Corresponding author)

Onderzoeksoutput: Bijdrage aan tijdschriftTijdschriftartikelAcademicpeer review

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Samenvatting

Characterization of the number and distribution of biological molecules on 2D surfaces is of foremost importance in biology and biomedicine. Synthetic surfaces bearing recognition motifs are a cornerstone of biosensors, while receptors on the cell surface are critical/vital targets for the treatment of diseases. However, the techniques used to quantify their abundance are qualitative or semi-quantitative and usually lack sensitivity, accuracy, or precision. Detailed herein a simple and versatile workflow based on super-resolution microscopy (DNA-PAINT) was standardized to improve the quantification of the density and distribution of molecules on synthetic substrates and cell membranes. A detailed analysis of accuracy and precision of receptor quantification is presented, based on simulated and experimental data. We demonstrate enhanced accuracy and sensitivity by filtering out non-specific interactions and artifacts. While optimizing the workflow to provide faithful counting over a broad range of receptor densities. We validated the workflow by specifically quantifying the density of docking strands on a synthetic sensor surface and the densities of PD1 and EGF receptors (EGFR) on two cellular models.
Originele taal-2Engels
Pagina's (van-tot)80-93
Aantal pagina's14
TijdschriftACS Sensors
Volume8
Nummer van het tijdschrift1
Vroegere onlinedatum19 jan. 2023
DOI's
StatusGepubliceerd - 27 jan. 2023

Bibliografische nota

Publisher Copyright:
© 2023 The Authors. Published by American Chemical Society.

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