Development of an inexpensive Raman-compatible substrate for the construction of a microarray screening platform
| Autor: | Isamar Pastrana-Otero, Brendan A.C. Harley, Brittney L. Gorman, Mary L. Kraft, Aidan E. Gilchrist, Sayani Majumdar |
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| Rok vydání: | 2020 |
| Předmět: |
Microarray
Substrate (printing) Spectrum Analysis Raman 01 natural sciences Biochemistry Article Analytical Chemistry 03 medical and health sciences symbols.namesake Mice Cricetulus Tissue engineering Single-cell analysis Cricetinae Electrochemistry Environmental Chemistry Animals Least-Squares Analysis Spectroscopy 030304 developmental biology 0303 health sciences Chemistry 010401 analytical chemistry Biomaterial Discriminant Analysis Cell Differentiation Transfection 0104 chemical sciences symbols Stem cell DNA microarray Raman spectroscopy Adult stem cell Biomedical engineering |
| Zdroj: | Analyst |
| ISSN: | 1364-5528 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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