Semi-automated quantification of living cells with internalized nanostructures

Journal of Nanobiotechnology
Michael Bogdan MargineanuTimothy Ravasi

Abstract

Nanostructures fabricated by different methods have become increasingly important for various applications in biology and medicine, such as agents for medical imaging or cancer therapy. In order to understand their interaction with living cells and their internalization kinetics, several attempts have been made in tagging them. Although methods have been developed to measure the number of nanostructures internalized by the cells, there are only few approaches aimed to measure the number of cells that internalize the nanostructures, and they are usually limited to fixed-cell studies. Flow cytometry can be used for live-cell assays on large populations of cells, however it is a single time point measurement, and does not include any information about cell morphology. To date many of the observations made on internalization events are limited to few time points and cells. In this study, we present a method for quantifying cells with internalized magnetic nanowires (NWs). A machine learning-based computational framework, CellCognition, is adapted and used to classify cells with internalized and no internalized NWs, labeled with the fluorogenic pH-dependent dye pHrodo™ Red, and subsequently to determine the percentage of cells with ...Continue Reading

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Citations

Oct 25, 2016·Scientific Reports·Aldo Isaac Martínez-BanderasJürgen Kosel
May 12, 2018·International Journal of Molecular Sciences·Łukasz OpalińskiJacek Otlewski
Mar 14, 2020·Journal of Nanobiotechnology·Aldo Isaac Martínez-BanderasJürgen Kosel
Oct 29, 2020·Journal of Applied Toxicology : JAT·Eşref Demir
Dec 29, 2020·Journal of Colloid and Interface Science·Laura I FitzGerald, Angus P R Johnston
Nov 5, 2019·ACS Applied Materials & Interfaces·Aldo Isaac Martínez BanderasJurgen Kosel

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Methods Mentioned

BETA
confocal microscopy
infrared spectroscopy
flow
fluorescence activated cell sorting
FACS
flow cytometry
feature extraction

Software Mentioned

CellCognition
CellCogniton
FACS
CecogAnalyzer
Image J
FlowJo

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