4.8 Article

Expansion Microscopy with a Thermally Adjustable Expansion Factor Using Thermoresponsive Biospecimen-Hydrogel Hybrids

期刊

ACS APPLIED MATERIALS & INTERFACES
卷 13, 期 24, 页码 28962-28974

出版社

AMER CHEMICAL SOC
DOI: 10.1021/acsami.1c07592

关键词

expansion microscopy; cell-hydrogel hybrid; super-resolution microscopy; adjustable expansion factor; thermoresponsive hydrogel

资金

  1. Samsung Research Funding & Incubation Center of Samsung Electronics [SRFC-MA1601-08]

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T-RevExM is a technique that allows for adjusting the size of biological samples by controlling temperature, effectively increasing the magnification and resolution of the sample.
Expansion microscopy (ExM) is a technique in which swellable hydrogel-embedded biological samples are physically expanded to effectively increase imaging resolution. Here, we develop thermoresponsive reversible ExM (T-RevExM), in which the expansion factor can be thermally adjusted in a reversible manner. In this method, samples are embedded in thermoresponsive hydrogels and partially digested to allow for reversible swelling of the sample-gel hybrid in a temperature-dependent manner. We first synthesized hydrogels exhibiting lower critical solution temperature (LCST)- and upper critical solution temperature (UCST)-phase transition properties with N-alkyl acrylamide or sulfobetaine monomers, respectively. We then formed covalent hybrids between the LCST or UCST hydrogel and biomolecules across the cultured cells and tissues. The resulting hybrid could be reversibly swelled or deswelled in a temperature-dependent manner, with LCST- and UCST-based hybrids negatively and positively responding to the increase in temperature (termed thermonegative RevExM and thermopositive RevExM, respectively). We further showed reliable imaging of both unexpanded and expanded cells and tissues and demonstrated minimal distortions from the original sample using conventional confocal microscopy. Thus, T-RevExM enables easy adjustment of the size of biological samples and therefore the effective magnification and resolution of the sample, simply by changing the sample temperature.

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