Real-Time Chiral Metabolic Monitoring of Single Cell Using Microchip Electrophoresis Coupled with Electrospray Ionization Mass Spectrometry

摘要

Cells are extremely complex and dynamic biochemical entities. Significant biochemical heterogeneity can exist among cells of the same type. This heterogeneity may arise via many mechanisms, including localized damage, mutations, stages in the cell cycle, and differential exposure to external signals among others. ~[1,2] Unfortunately, when working with measure- ment approaches that report average values for larger numbers of cells, critical information is often not obtained. ~[3] To under- standing the chemical differences between cell types, the techniques that can examine only one cell at a time (namely single cell analysis) are necessary. ~[4] Single cell analysis can provide an insight to the detailed chemistry information of a cell on individual, which allows exploration of the cells and the relationship to their overall pathological mechanism. However, single cell analysis is an emerging field requiring a high level interdisciplinary collaboration to provide detailed insights into the complex organisation, function and heterogeneity of life. And the cells are generally on the order of 5–20 mmin diameter. Many of the macromolecules of interest are present in low copy numbers, so the techniques which can handle such small volumes without significant dilution and which can be directly integrated with ultrasensitive detection schemes are required. Single-cell methods for genome sequence, ~[5,6] tran- scrip-tomes, small-volume analysis of cell-cell signaling molecules, [12]~ metabolites, [13, 14]~ cell fates, ~[15] DNA methyla-tion, ~[16] yeast proteome dynamics, ~[17] mass spectrometry (MS) imag- ing, ~[18, 19] and chromosome conformation~[20] have been reported. However, the technologies for real-time chiral metabolic monitoring in single cell are still a great challenge.