Identifying Individual Cell Types in Heterogeneous Cultures Using Secondary Ion Mass Spectrometry Imaging with C60 Etching and Multivariate Analysis

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• 作者：Christopher A. Barnes ; Jeremy Brison ; Michael Robinson ; Daniel J. Graham ; David G. Castner ; Buddy D. Ratner
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：January 17, 2012
• 年：2012
• 卷：84
• 期：2
• 页码：893-900
• 全文大小：690K
• 年卷期：v.84,no.2(January 17, 2012)
• ISSN：1520-6882

文摘

Tissue engineering approaches fabricate and subsequently implant cell-seeded and unseeded scaffold biomaterials. Once in the body, these biomaterials are repopulated with somatic cells of various phenotypes whose identification upon explantation can be expensive and time-consuming. We show that imaging time-of-flight secondary ion mass spectrometry (TOF-SIMS) can be used to distinguish mammalian cell types in heterogeneous cultures. Primary rat esophageal epithelial cells (REEC) were cultured with NIH 3T3 mouse fibroblasts on tissue culture polystyrene and freeze-dried before TOF-SIMS imaging. Results show that a short etching sequence with C₆₀⁺ ions can be used to clean the sample surface and improve the TOF-SIMS image quality. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were used to identify peaks whose contributions to the total variance in the multivariate model were due to either the two cell types or the substrate. Using PLS-DA, unknown regions of cellularity that were otherwise unidentifiable by SIMS could be classified. From the loadings in the PLS-DA model, peaks were selected that were indicative of the two cell types and TOF-SIMS images were created and overlaid that showed the ability of this method to distinguish features visually.