Science and Research

Development of a Gas-Tight Microfluidic System for Raman Sensing of Single Pulmonary Arterial Smooth Muscle Cells Under Normoxic/Hypoxic Conditions

Acute hypoxia changes the redox-state of pulmonary arterial smooth muscle cells (PASMCs). This might influence the activity of redox-sensitive voltage-gated K(+)-channels (Kv-channels) whose inhibition initiates hypoxic pulmonary vasoconstriction (HPV). However, the molecular mechanism of how hypoxia-or the subsequent change in the cellular redox-state-inhibits Kv-channels remains elusive. For this purpose, a new multifunctional gas-tight microfluidic system was developed enabling simultaneous single-cell Raman spectroscopic studies (to sense the redox-state under normoxic/hypoxic conditions) and patch-clamp experiments (to study the Kv-channel activity). The performance of the system was tested by optically recording the O(2)-content and taking Raman spectra on murine PASMCs under normoxic/hypoxic conditions or in the presence of H(2)O(2). Oxygen sensing showed that hypoxic levels in the gas-tight microfluidic system were achieved faster, more stable and significantly lower compared to a conventional open system (1.6 +/- 0.2%, respectively 6.7 +/- 0.7%, n = 6, p < 0.001). Raman spectra revealed that the redistribution of biomarkers (cytochromes, FeS, myoglobin and NADH) under hypoxic/normoxic conditions were improved in the gas-tight microfluidic system (p-values from 0.00% to 16.30%) compared to the open system (p-value from 0.01% to 98.42%). In conclusion, the new redox sensor holds promise for future experiments that may elucidate the role of Kv-channels during HPV.
  • Knoepp, F.
  • Wahl, J.
  • Andersson, A.
  • Borg, J.
  • Weissmann, N.
  • Ramser, K.

Keywords

  • Raman spectroscopy
  • hypoxia
  • microfluidic system
  • redox reactions on single cell level
Publication details
DOI: 10.3390/s18103238
Journal: Sensors (Basel, Switzerland)
Number: 10
Work Type: Original
Location: UGMLC
Disease Area: General Lung and Other
Partner / Member: JLU
Access-Number: 30261634
See publication on PubMed

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