Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.
Department of Pharmacology, University of Michigan, Ann Arbor, MI, 48109, USA.
Anal Bioanal Chem. 2023 Feb;415(4):533-544. doi: 10.1007/s00216-022-04460-2. Epub 2022 Dec 2.
Tissue functions such as hormone secretion involve the interplay of multiple chemical signals and metabolic processes over time. Measuring the different components involved is useful in unraveling the interactions, but often requires use of multiple analytical techniques. The challenge of measuring the necessary components with temporal resolution is greater when tissue samples are limited. Here, an accessible microfluidic platform compatible with multiple measurement techniques to monitor cell secretions has been developed. The platform is applied to islets of Langerhans, micro-organs involved in glucose homeostasis and diabetes. The device houses 1 to 8 islets and the perfusion fluid can be controlled to change conditions, e.g., glucose concentration, in seconds. Samples are collected in fractions and split for offline analysis. The device is paired with a scaled-down immunoassay, AlphaLISA, for hormone quantification and liquid chromatography-mass spectrometry for small molecule quantification to study secretion dynamics. The combined system allows the first simultaneous measurement of insulin, glucagon, biogenic amines, and amino acids from islet secretions. The combined measurements revealed correlation in secretion events and differences in timing of release between hormones and biogenic amines and amino acids. These efforts decreased the number of islets required compared to standard approaches, thus decreasing necessary animal use, reagent use, and cost, while increasing information content achievable from one sample. The microfluidic device is a suitable platform for in-depth characterization of secretion from small tissue samples.
组织的功能，如激素分泌，涉及多种化学信号和代谢过程随时间的相互作用。测量涉及的不同成分对于揭示相互作用是有用的，但通常需要使用多种分析技术。当组织样本有限时，具有时间分辨率测量必要成分的挑战更大。在这里，开发了一种可与多种测量技术兼容的易于使用的微流控平台，以监测细胞分泌物。该平台应用于参与葡萄糖稳态和糖尿病的胰岛。该设备容纳 1 到 8 个胰岛，并且可以控制灌注液在几秒钟内改变条件，例如葡萄糖浓度。样品以分数形式收集并分开进行离线分析。该设备与缩小规模的免疫分析（AlphaLISA）配对，用于激素定量，以及液相色谱-质谱法用于小分子定量，以研究分泌动力学。该组合系统允许首次同时测量胰岛分泌的胰岛素、胰高血糖素、生物胺和氨基酸。联合测量显示了激素和生物胺以及氨基酸分泌事件之间的相关性和释放时间的差异。与标准方法相比，这些努力减少了所需的胰岛数量，从而减少了所需的动物使用、试剂使用和成本，同时增加了从一个样本中获得的信息量。微流控设备是从小组织样本中深入表征分泌的合适平台。
