A multi-functional flow & patch chamber system for cell research and patch clamp applications

Journal of Pharmacological and Toxicological Methods, vol. 140, pp. 108535

Abstract

AI applications has established itself as a powerful tool for cell biology research in a new, quantitative dimension. We present five cell research AI applications in combination with a microfluidic device, and evaluate the results. Additionally, new potential avenues for patch clamp research are highlighted. Methodologically, microscopic observations are analyzed with specific AI solutions and compared with results from conventional technology. Applications: 1) stiffness of red blood cells (RBCs) including quantitative subpopulation analysis, 2) shear stress effect (< 3 Pa) on blood cell endothelial cell (EC) adhesion, 3) osmotic fragility of healthy RBCs (potentially in hemolytic anemias), 4) healthy induced RBC aggregate formation (potential tool for inflammation research) and 5) effect of temperature, shear flow and drugs in patch clamp measurements on the same measured cell. Results: 1) AI-RBC stiffness experiments showed that glutaraldehyde (0.1%, 15 min, Hct 2.5%) results in a significant stiffening of 8% compared to untreated healthy RBCs and no subpopulation forming, 2) The mean AI-derived cell count of RBCs adhering to ECs following two minutes of fluid shear at 0.25 Pa demonstrated a 98.7% agreement with visual counting. 3) the reference range for MCF 50 (mean corpuscular fragility) for classic osmotic fragility methods was established as 0.40–0.45%, whilst the AI-based range was 0.39–0.45%. The AI approach is suggested for investigating haemolytic anaemia patients, offering unparalleled simplicity. 4) the number of RBC roleaux formation induced by increased plasma with RBCs resulted in numbers identical to those of manually counted ones. 5) patch clamp evaluation studies are still ongoing. They are performed within 20 °C to 45 °C at ±0.2 °C accuracy and at a shear flow of 0.25 Pa around the patched cell. Precise data on critical switching temperatures of TRP channels or enzymes where Q10 values switch are expected. The above examples demonstrate the high potential of similar AI approaches for further applications, e.g., dose-response characteristics, substance wash-in and wash-out times, as well as other in vitro cell studies of sickle cell disease, thalassemia, coronavirus studies (SARS-CoV-2), white blood cell activation, or endothelial responses to mechanical stimuli.

Authors 4

1. Özgür Alaçayır Aachen Center of Competence for Bioengineering

   FH Aachen

   Affiliation as printed

   Center of Competence for Bioengineering, University of Applied Sciences Aachen, Medical and Biological Laboratory, Jülich, NRW, Germany

2. Till Peter Creutz Aachen Center of Competence for Bioengineering

   FH Aachen · Ministerium für Verkehr des Landes Nordrhein-Westfalen

   Affiliation as printed

   Center of Competence for Bioengineering, University of Applied Sciences Aachen, Medical and Biological Laboratory, Jülich, NRW, Germany

   HiTec Zang GmbH, Herzogenrath, NRW, Germany

3. G.M. Artmann Aachen Center of Competence for Bioengineering

   FH Aachen

   Affiliation as printed

   Center of Competence for Bioengineering, University of Applied Sciences Aachen, Medical and Biological Laboratory, Jülich, NRW, Germany

4. Ayşegül Temiz Artmann Aachen Center of Competence for Bioengineering

   FH Aachen

   Affiliation as printed

   Center of Competence for Bioengineering, University of Applied Sciences Aachen, Medical and Biological Laboratory, Jülich, NRW, Germany