Fluorescent Hemolysis Detection (FHD): Vaidation of the in-vitro test method
Final Report Abstract
We have investigated fluorescent hemolysis detection (FHD) in a two-phase blood analogue fluid consisting of ghost cells (GCs) and plasma. In FHD, an indicator (Cal-590™ potassium salt, AAT Bioquest, Pleasanton, California) is separated from its target (calcium ions) by a cell membrane (GCs). When the cell membrane is lysed, the two bind, resulting in a fluorescent signal that is detected optically with spatial and temporal resolution. In the project, we characterized the blood-analogue fluid and further developed the FDH method to enable spatially resolved hemolysis detection in medical devices. Standardized hemolysis tests compare two test circuits of less than 500 mL blood volume each. In order to produce GCs in this amount, a large-scale production of GCs was realized in the previous project. However, due to an error in the hematocrit measurement, the quantity of GCs produced was overestimated, so that the production process had to be further developed in order to be able to produce a larger quantity of GCs. Now, at least 350 mL of pure GCs can be reproducibly produced, from which 1,000 mL of blood analogue fluid can be produced with plasma, which has a hematocrit (HCT) of 35 % and can be used in a comparable way to standardized hemolysis tests. To quantify the FHD signal in terms of hemolysis of the GCs, different hemolysis hotspots were used to induce controlled hemolysis and FHD was performed optically. The hemolysis hotspots were a microchannel with high pressure and flow and a static reaction vessel exposed to ultrasonic shock waves. Significant cell damage was detected in both experiments. However, a FHD signal could not be detected during the evaluation of the video monitoring of the hemolysis hotspots. Instead, fluid samples taken during the experiment were optically analyzed for an FHD signal. In a spatially resolved FHD measurement, all generated results are to be combined in which hemolysis in the FDA pump is displayed spatially resolved.
Publications
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Conference Presentation and Poster, The International Society for Mechanical Circulatory Support, Hannover: Visualize Mechanical Hemolysis with Ghost Cells – A Feasibility Study for Hemolysis Detection 2021 (postponed to 2022)
Benjamin J. Schürmann, Eugenia Weber, Claudio A. Luisi, Ilona Mager, Thomas Schmitz-Rode, Ulrich Steinseifer & Johanna C. Clauser
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Conference Presentation, European Society for Artificial Organs, Krems: Optical Visualization of Spatially Resolved Mechanical Hemolysis by Means of Ghost Cells 2022
Benjamin J. Schürmann, Eugenia Weber, Claudio A. Luisi, Ilona Mager, Sebastian V. Jansen, Thomas Schmitz-Rode, Ulrich Steinseifer & Johanna C. Clauser
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Conference Poster, European Society for Artificial Organs, Bergamo: Optical Analysis of Ghost Cells under Mechanical Hemolysis using Fluorescence Hemolysis Detection 2023
Benjamin J. Schürmann, Lavanja Arunthavarasa, Nina Reinhardt, Pia Hefer, Ilona Mager, Sebastian V. Jansen, Thomas Schmitz-Rode, Ulrich Steinseifer & Johanna C. Clauser
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Conference Presentation, Blood Damage Workshop, Rostock: Ghost Cells as a Transparent Blood Substitute Fluid – Microscopic Examination of Cell Shape, Size and Deformability 2023
Benjamin J. Schürmann, Miriam D. Mineur, Lucas Stüwe, Pia Hefer, Thomas Schmitz-Rode, Ulrich Steinseifer & Johanna C. Clauser
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Conference Presentation, European Society for Artificial Organs, Bergamo: Ghost Blood- A Novel Fluid for Visual Monitoring of Coagulation in an Occlusion System 2023
P. Hefer, B. J. Schuermann, C. Luisi, X. Yin, U. Steinseifer, A. Sedaghat & J. C. Clauser
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Ghost cells as a two-phase blood analog fluid —fluorescent mechanical hemolysis detection. Springer Science and Business Media LLC.
Schürman, Benjamin J.; Holst, Bennet F.; Creutz, Pia; Schmitz-Rode, Thomas; Steinseifer, Ulrich & Clauser, Johanna C.
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Ghost cells as a two‐phase blood analog fluid: high‐volume and high‐concentration production. Artificial Organs, 49(1), 31-38.
Schürmann, Benjamin J.; Creutz, Pia; Schmitz‐Rode, Thomas; Steinseifer, Ulrich & Clauser, Johanna C.
