Development of an inexpensive Raman-compatible substrate for the construction of a microarray screening platform.
Development of an inexpensive Raman-compatible substrate for the construction of a microarray screening platform.
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DOI：
10.1039/d0an01153c
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发表时间：
2020-10-26
期刊：
影响因子：
--
通讯作者：
Kraft ML
中科院分区：
文献类型：
--
作者：
Pastrana-Otero I ;Majumdar S ;Gilchrist AE ;Gorman BL ;Harley BAC ;Kraft ML
生物材料微阵列正在开发，以促进识别外部线索，引起干细胞的命运决定自我更新，分化和保持静止。拉曼显微光谱，通常与多变量分析技术，如偏最小二乘判别分析（PLS-DA）相结合，可以使非侵入性的识别干细胞的命运决定，在这些微阵列上的特定位置的外在线索。由于现有的生物材料微阵列与拉曼显微光谱不兼容，在这里，我们开发了一种廉价的基板，它与单细胞拉曼光谱和通常用于生物材料微阵列制造的化学物质兼容。使用标准沉积技术来制造支持微阵列构建的定制拉曼相容基底。我们验证了从附接至定制拉曼兼容基底的功能化聚丙烯酰胺（PA）凝胶上的活细胞获得的光谱与从更昂贵的市售基底获得的光谱相当。我们还表明，从连接到我们的定制基板上的功能化PA凝胶上的单个活细胞获得的光谱具有足够的质量，能够使用细胞光谱的PLS-DA模型准确鉴定细胞表型。我们通过使用来自实验室系的细胞（CHO和转染的CHO细胞）以及从小鼠新鲜分离的成体干细胞（长期和短期造血干细胞）来证明这一点。本文报道的定制拉曼相容性基底可用作构建生物材料微阵列的廉价基底，所述生物材料微阵列使得能够使用拉曼显微光谱法来非侵入性地鉴定干细胞响应于外在线索的命运决定。
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 Raman-compatible 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.
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影响因子：
7.4
作者：
Ilin Y;Choi JS;Harley BA;Kraft ML
通讯作者：
Kraft ML
影响因子：
3.8
作者：
Rangan, Shreyas;Kamal, Sepehr;Piret, James M.
通讯作者：
Piret, James M.
影响因子：
2.4
作者：
Musumarra, G;Barresi, V;Scirè, S
通讯作者：
Scirè, S
影响因子：
2.5
作者：
Downes, Andrew;Mouras, Rabah;Bagnaninchi, Pierre;Elfick, Alistair
通讯作者：
Elfick, Alistair
影响因子：
3.7
作者：
Challen, Grant A.;Boles, Nathan;Lin, Karen Kuan-Yin;Goodell, Margaret A.
通讯作者：
Goodell, Margaret A.