Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States
Catherine M Della Santina
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States
Pema Maretich
Department of Biology, Massachusetts Institute of Technology, Cambridge, United States
Alexander L Auld
Department of Biology, Massachusetts Institute of Technology, Cambridge, United States
Kirsten K Schnieder
Department of Biology, Massachusetts Institute of Technology, Cambridge, United States
Tay Shin
Department of Media Arts and Sciences, Massachusetts Institute of Technology, Cambridge, United States
Edward S Boyden
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States; Department of Media Arts and Sciences, Massachusetts Institute of Technology, Cambridge, United States; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, United States; McGovern Institut, Massachusetts Institute of Technology, Cambridge, United States; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, United States; K Lisa Yang Center for Bionics, Massachusetts Institute of Technology, Cambridge, United States; Center for Neurobiological Engineering, Massachusetts Institute of Technology, Cambridge, United States; Koch Institute, Massachusetts Institute of Technology, Cambridge, United States
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States; Department of Biology, Massachusetts Institute of Technology, Cambridge, United States; Koch Institute, Massachusetts Institute of Technology, Cambridge, United States
Expansion microscopy (ExM) enables nanoscale imaging using a standard confocal microscope through the physical, isotropic expansion of fixed immunolabeled specimens. ExM is widely employed to image proteins, nucleic acids, and lipid membranes in single cells; however, current methods limit the number of samples that can be processed simultaneously. We developed High-throughput Expansion Microscopy (HiExM), a robust platform that enables expansion microscopy of cells cultured in a standard 96-well plate. Our method enables ~4.2 x expansion of cells within individual wells, across multiple wells, and between plates. We also demonstrate that HiExM can be combined with high-throughput confocal imaging platforms to greatly improve the ease and scalability of image acquisition. As an example, we analyzed the effects of doxorubicin, a known cardiotoxic agent, on human cardiomyocytes (CMs) as measured by the Hoechst signal across the nucleus. We show a dose-dependent effect on nuclear DNA that is not observed in unexpanded CMs, suggesting that HiExM improves the detection of cellular phenotypes in response to drug treatment. Our method broadens the application of ExM as a tool for scalable super-resolution imaging in biological research applications.
