Chang, Y.
, Kim, S.
and Lee, Y.
(2024),
Benchtop IR Imaging of Live Cells: Monitoring the Total Mass of Biomolecules in Single Cells, Analytical Chemistry, [online], https://doi.org/10.1021/acs.analchem.4c02108, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=935425
(Accessed December 30, 2024)
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Benchtop IR Imaging of Live Cells: Monitoring the Total Mass of Biomolecules in Single Cells
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Abstract
Absolute quantity imaging of biomolecules on a single cell level is critical for measurement assurance in biosciences and bioindustries. While infrared (IR) transmission microscopy is a powerful label-free imaging modality capable of chemical quantification, its applicability to hydrated biological samples remains challenging due to the strong IR absorption by water. Traditional IR imaging of hydrated cells relies on powerful light sources, such as synchrotrons, to mitigate the light absorption by water. However, we overcome this challenge by applying a solvent absorption compensation (SAC) technique to a home-built benchtop IR microscope based on an external-cavity quantum cascade laser. SAC-IR microscopy adjusts the incident light using a pair of polarizers to pre-compensate the IR absorption by water while retaining the full dynamic range. Integrating the IR absorbance over a cell yields the total mass of biomolecules per cell. We monitor the total mass of the biomolecules of live fibroblast cells over twelve hours, demonstrating promise for advancing our understanding of the biomolecular processes occurring in live cells on the single-cell level.Citation
Analytical Chemistry
Volume
96
Issue
37
Pub Type
Journals
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Keywords
IR absorption, label-free chemical imaging, live cell imaging
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Created September 4, 2024, Updated December 19, 2024