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FRET Imaging in Three-dimensional Hydrogels
Published on: August 1, 2016
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SERS-Active Printable Hydrogel for 3D Cell Culture and Imaging
Wei Wang1,2, Peter J Vikesland1,2
1Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, Virginia 24061, United States.
Analytical Chemistry
|November 7, 2023
Summary
Researchers developed 3D-printable SERS-active hydrogels using gold nanoparticles for cell imaging. This technique allows label-free visualization of cellular components and behavior within 3D cultures.
Area of Science:
- Biomaterials Engineering
- Spectroscopy
- Cell Biology
Background:
- Three-dimensional (3D) cell culture systems are crucial for mimicking native tissue environments.
- High-resolution 3D cell imaging is needed to capture detailed cellular information.
- Current imaging techniques often lack the resolution or specificity for complex 3D cultures.
Purpose of the Study:
- To develop a novel 3D cell imaging method using surface-enhanced Raman spectroscopy (SERS).
- To create SERS-active hydrogel inks for 3D printing and culturing of cells.
- To enable label-free, high-resolution imaging of cellular components and behavior in 3D.
Main Methods:
- Incorporation of plasmonic gold nanoparticles (AuNPs) into alginate/gelatin hydrogels.
- 3D printing of SERS-active hydrogel inks for cell culture.
- Label-free SERS spectral acquisition within the hydrogel.
- Development of a random forest classifier to distinguish cell signals from background.
- Image reconstruction combining classification and intensity maps for 2D and 3D visualization.
Main Results:
- Successfully fabricated 3D SERS-active hydrogel structures with no adverse effects on cell growth.
- Achieved 87.5% accuracy in discriminating cell signals from hydrogel background using a classifier.
- Demonstrated visualization of cell shape, location, and density with ≈3 μm resolution.
- Observed increased SERS signal intensity with incubation time, correlating with cell behavior.
Conclusions:
- 3D SERS cell imaging is a viable technique for high-resolution, label-free analysis of cells in 3D cultures.
- This method allows for the examination of cellular components (proteins, lipids, carbohydrates) and their spatial distribution.
- The developed hydrogel inks and imaging approach hold promise for diagnostic applications and studying cellular responses to stimuli.
