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CC BY 4.0 · Thromb Haemost 2025; 125(04): 340-351
DOI: 10.1055/a-2378-9088
Cellular Haemostasis and Platelets

Volume Regulation and Nonosmotic Volume of Individual Human Platelets Quantified by High-Speed Scanning Ion Conductance Microscopy

Konstantin Krutzke 
1   Institute of Applied Physics, University of Tübingen, Tübingen, Germany
,
Jan Seifert 
1   Institute of Applied Physics, University of Tübingen, Tübingen, Germany
,
Meinrad Gawaz 
2   Department of Internal Medicine III, Cardiology and Angiology, University of Tübingen, Tübingen, Germany
,
Johannes Rheinlaender 
1   Institute of Applied Physics, University of Tübingen, Tübingen, Germany
,
Tilman E. Schäffer 
1   Institute of Applied Physics, University of Tübingen, Tübingen, Germany
› Author Affiliations
Funding This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—Projektnummer 335549539/GRK2381 and Projektnummer 374031971-TRR 240. We acknowledge support by Open Access Publishing Fund of the University of Tübingen.
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Abstract

Background

Platelets are anucleate cells that play an important role in wound closure following vessel injury. Maintaining a constant platelet volume is critical for platelet function. For example, water-induced swelling can promote procoagulant activity and initiate thrombosis. However, techniques for measuring changes in platelet volume such as light transmittance or impedance techniques have inherent limitations as they only allow qualitative measurements or do not work on the single-cell level.

Methods

Here, we introduce high-speed scanning ion conductance microscopy (HS-SICM) as a new platform for studying volume regulation mechanisms of individual platelets. We optimized HS-SICM to quantitatively image the morphology of adherent platelets as a function of time at scanning speeds up to 7 seconds per frame and with 0.1 fL precision.

Results

We demonstrate that HS-SICM can quantitatively measure the rapid swelling of individual platelets after a hypotonic shock and the following regulatory volume decrease (RVD). We found that the RVD of thrombin-, ADP-, and collagen-activated platelets was significantly reduced compared with nonactivated platelets. Applying the Boyle–van't Hoff relationship allowed us to extract the nonosmotic volume and volume fraction on a single-platelet level. Activation by thrombin or ADP, but not by collagen, resulted in a decrease of the nonosmotic volume, likely due to a release reaction, leaving the total volume unaffected.

Conclusion

This work shows that HS-SICM is a versatile tool for resolving rapid morphological changes and volume dynamics of adherent living platelets.

Keywords

hypotonic shock - platelet volume - nonosmotic fraction - platelet activation - SICM

Authors' Contribution

K.K., J.S., M.G., J.R., and T.S. designed the study. K.K. performed the experiments, analyzed data, and drafted the manuscript. All authors interpreted data, discussed results, and revised the manuscript.


Supplementary Material



Publication History

Received: 21 December 2023

Accepted: 28 July 2024

Accepted Manuscript online:
02 August 2024

Article published online:
29 August 2024

© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

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