AUTHOR=Yang Boxuan , Richards Ceri J. , Gandek Timea B. , de Boer Isa , Aguirre-Zuazo Itxaso , Niemeijer Else , Åberg Christoffer TITLE=Following nanoparticle uptake by cells using high-throughput microscopy and the deep-learning based cell identification algorithm Cellpose JOURNAL=Frontiers in Nanotechnology VOLUME=5 YEAR=2023 URL=https://www.frontiersin.org/journals/nanotechnology/articles/10.3389/fnano.2023.1181362 DOI=10.3389/fnano.2023.1181362 ISSN=2673-3013 ABSTRACT=

How many nanoparticles are taken up by human cells is a key question for many applications, both within medicine and safety. While many methods have been developed and applied to this question, microscopy-based methods present some unique advantages. However, the laborious nature of microscopy, in particular the consequent image analysis, remains a bottleneck. Automated image analysis has been pursued to remedy this situation, but offers its own challenges. Here we tested the recently developed deep-learning based cell identification algorithm Cellpose on fluorescence microscopy images of HeLa cells. We found that the algorithm performed very well, and hence developed a workflow that allowed us to acquire, and analyse, thousands of cells in a relatively modest amount of time, without sacrificing cell identification accuracy. We subsequently tested the workflow on images of cells exposed to fluorescently-labelled polystyrene nanoparticles. This dataset was then used to study the relationship between cell size and nanoparticle uptake, a subject where high-throughput microscopy is of particular utility.