4.8 Article

SERS-Active Printable Hydrogel for 3D Cell Culture and Imaging

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ANALYTICAL CHEMISTRY
卷 95, 期 49, 页码 18055-18064

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AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.3c02641

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This study incorporates gold nanoparticles into a hydrogel ink for 3D printing and culturing of cells, enabling the use of surface-enhanced Raman spectroscopy (SERS) for cell imaging. A random forest binary classifier is developed to accurately distinguish cell signals from the background. The results show that SERS signals provide information about cellular components.
Hydrogel-based three-dimensional (3D) cell culture systems mimic the salient elements of extracellular matrices and promote native cell function. However, high-resolution 3D cell imaging that can provide biological information about multiple features of individual cells is yet to be realized. In this context, we incorporated plasmonic gold nanoparticles (AuNPs) into an alginate/gelatin hydrogel to produce surface-enhanced Raman spectroscopy (SERS)-active hydrogel inks for the 3D printing and culturing of Vero cells. Dense incorporation of AuNPs enables production of a printed 3D grid structure with 3D SERS performance, but with no measurable adverse effects on cell growth. Label-free SERS spectra were collected within a hydrogel, and a random forest binary classifier was developed to discriminate Vero cell signals from the hydrogel background with an accuracy of 87.5%. The results suggest that SERS signals from cellular components, such as proteins, lipids, and carbohydrates, account for this discrimination. We demonstrate visualization of cell shape, location, and density by combining predicted binary maps with peak feature intensity maps in 2D and 3D. SERS images with a resolution of approximate to 3 mu m match well with the microscopy images and show clear increases in intensity with incubation time. We suggest that 3D SERS cell imaging is a promising means to examine the effect of external cell stimuli on cellular behavior for diagnostic purposes.

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