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Development of an Inexpensive Raman-Compatible Substrate for the Construction of a Microarray Screening Platform

preprint
submitted on 09.06.2020 and posted on 12.06.2020 by Isamar Pastrana-Otero, Sayani Majumdar, Aidan E. Gilchrist, Brittney L. Gorman, Brendan A. C. Harley, Mary L. Kraft
Biomaterial microarrays are being developed to facilitate identifying the extrinsic cues that elicit stem cell fate decisions to self-renew, differentiate and remain quiescent. Raman microspectroscopy, often combined with multivariate analysis techniques such as partial least square-discriminant analysis (PLS-DA), could enable the non-invasive identification of stem cell fate decisions made in response to extrinsic cues presented at specific locations on these microarrays. Because existing biomaterial microarrays are not compatible with Raman microspectroscopy, here, we develop an inexpensive substrate that is compatible with both single-cell Raman spectroscopy and the chemistries that are often used for biomaterial microarray fabrication. Standard deposition techniques were used to fabricate a custom Raman-compatible substrate that supports microarray construction. We validated that spectra from living cells on functionalized polyacrylamide (PA) gels attached to the custom Raman-compatible substrate are comparable to spectra acquired from a more expensive commercially available substrate. We also showed that the spectra acquired from individual living cells on functionalized PA gels attached to our custom substrates were of sufficient quality to enable accurate identification of cell phenotypes using PLS-DA models of the cell spectra. We demonstrated this by using cells from laboratory lines (CHO and transfected CHO cells) as well as adult stem cells that were freshly isolated from mice (long-term and short-term hematopoietic stem cells). The custom Ramancompatible substrate reported herein may be used as an inexpensive substrate for constructing biomaterial microarrays that enable the use of Raman microspectroscopy to non-invasively identify the fate decisions of stem cells in response to extrinsic cues.

Funding

R21 HL132642

R01 CA197488

R01 DK099528

R21 EB018481

F31 DK117514

T32 EB019944

CHE-1508662

History

Email Address of Submitting Author

mlkraft@illinois.edu

Institution

University of Illinois at Urbana-Champaign

Country

United States of America

ORCID For Submitting Author

0000-0002-5879-7222

Declaration of Conflict of Interest

No competing financial interests exist.

Version Notes

Submitted version

Exports