Monitoring Intracellular Calcium in Response to GPCR Activation: Comparison Between Microtiter Plates and Microfluidic Assays
| Autor: | Duarte M. F. Prazeres, Virginia Chu, S. A. M. Martins, João Pedro Conde |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0303 health sciences
HEK 293 cells Microfluidics chemistry.chemical_element 02 engineering and technology Calcium 021001 nanoscience & nanotechnology Pirenzepine Calcium in biology 03 medical and health sciences Transduction (genetics) chemistry In vivo medicine Biophysics 0210 nano-technology 030304 developmental biology medicine.drug Calcium signaling |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071612200 |
| DOI: | 10.1007/978-1-0716-1221-7_19 |
| Popis: | Microfluidic strategies combined with transduction and electronic integration have the promise of enabling miniaturized, combinatorial assays at higher speeds and lower costs, while at the same time mimicking the local chemical concentrations and force fields of the cellular in vivo environment. In this chapter we introduce a microfluidic structure with hydrodynamic cell traps and a culture volume in the nanoliter range (50 nL), to quantitatively evaluate the transient calcium response of the endogenous Muscarinic type 1 receptor (M1) in HEK 293 T cells. The microfluidic fabrication protocol is described as well as a methodology to monitor the cell response in real time, after stimulation with M1 agonists (e.g., carbachol) and antagonists (e.g., Pirenzepine). |
| Databáze: | OpenAIRE |
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