Václavek Tomáš, Foret František
Department of Bioanalytical Instrumentation, Institute of Analytical Chemistry of the CAS, Brno, Czech Republic.
Department of Biochemistry, Masaryk University, Brno, Czech Republic.
Electrophoresis. 2023 Jan;44(1-2):313-322. doi: 10.1002/elps.202100379. Epub 2022 Apr 18.
Analysis of cellular composition and metabolism at a single-cell resolution allows gaining more information about complex relationships of cells within tissues or whole living organisms by resolving the variance stemming from the cellular heterogeneity. Mass spectrometry (MS) is a perfect analytical tool satisfying the demanding requirements of detecting and identifying compounds present in such ultralow-volume samples of high chemical complexity. However, the method of sampling and sample ionization is crucial in obtaining relevant information. In this work, we present a microfluidic sampling platform that integrates single-cell extraction from MS-incompatible media with electrical cell lysis and nanoESI-MS analysis of human erythrocytes. Hemoglobin alpha and beta chains (300 amol/cell) were successfully identified in mass spectra of single-erythrocyte lysates.
在单细胞分辨率下对细胞组成和代谢进行分析，能够通过解析细胞异质性所产生的差异，获取更多关于组织或整个生物体中细胞复杂关系的信息。质谱（MS）是一种理想的分析工具，能够满足检测和鉴定存在于这种化学复杂性高的超低体积样品中化合物的严格要求。然而，采样和样品离子化方法对于获取相关信息至关重要。在这项工作中，我们展示了一种微流控采样平台，该平台将从与质谱不兼容的介质中提取单个细胞与电细胞裂解以及对人类红细胞进行纳升电喷雾电离质谱分析相结合。在单个红细胞裂解物的质谱图中成功鉴定出了血红蛋白α链和β链（300 阿托摩尔/细胞）。
