Raman imaging diagnosis of the early stage differentiation of mouse embryonic stem cell (mESC).

Raman microspectroscopy as a non-invasive and label-free technique was applied to diagnose the early stage differentiation of mouse embryonic stem cells. The differentiated and undifferentiated embryonic bodies (EBs) were cultured using handing drop method by the control of Leukemia Inhibitory Factor (LIF). Raman spectra of the periphery cells of differentiated EBs (PrE cells) and those of the interior of undifferentiated EBs (ES cells) were obtained to diagnose the stem cells of different differentiation. It was found from the spectra that the protein content increased as the cells differentiated. Principal component analysis (PCA) was carried out to further analyze the differences between ES cells and PrE cells. The first three principle components contained 98.19% from the total variance. Characteristic bands of ES and PrE cells were chosen to acquire Raman images of two cells according to the results of PCA. In the Raman images, PrE cells had a clear and bright outline in the peripheral areas while ES cells were difficult to identify, this could be a distinct characteristic to discriminate them. The result of the Raman images was consistent with the biological agreement that the differentiated cells were distributed around the periphery.