Accessibility navigation


Kinetx: a combined flow cytometry assay and analysis software framework to quantitatively measure and categorize platelet activation in real-time

Dunster, J., Mitchell, J., Mohammed, Y., Taylor, K. ORCID: https://orcid.org/0000-0002-4599-7727, Gibbins, J. ORCID: https://orcid.org/0000-0002-0372-5352 and Jones, C. ORCID: https://orcid.org/0000-0001-7537-1509 (2021) Kinetx: a combined flow cytometry assay and analysis software framework to quantitatively measure and categorize platelet activation in real-time. Journal of Visualized Experiments, 176. e62947. ISSN 1940-087X

[img]
Preview
Text (Open Access) - Published Version
· Available under License Creative Commons Attribution.
· Please see our End User Agreement before downloading.

987kB
[img] Text - Accepted Version
· Restricted to Repository staff only

1MB

It is advisable to refer to the publisher's version if you intend to cite from this work. See Guidance on citing.

To link to this item DOI: 10.3791/62947

Abstract/Summary

Platelets react rapidly to vascular injury and undergo activation in response to a range of stimuli to limit blood loss. Many platelet function tests measure endpoint responses after a defined time period and not the rate of platelet activation. However, the rate at which platelets convert extracellular stimuli into a functional response is an essential factor in determining how efficiently they can respond to injury, bind to a forming thrombus, and signal to recruit other platelets. This paper describes a flow cytometry-based platelet function assay that enables simultaneous data acquisition and sample stimulation and utilizes newly developed bespoke open-source software (Kinetx) to enable quantitative kinetic measurements of platelet granule release, fibrinogen binding, and intracellular calcium flux. Kinetix was developed in R so that users can alter parameters such as degree of smoothing, identification of outlying data points, or time scales. To aid users unfamiliar with the R environment, Kinetix analysis of data can be performed by a single command. Together, this allows real-time platelet activation metrics, such as rate, acceleration, time to peak-rate, time to peak-calcium, and qualitative shape changes, to be accurately and reproducibly measured and categorized. Kinetic measurements of platelet activation give a unique insight into platelets’ behavior during the first stages of activation and may provide a method of predicting the recruitment of platelets into a forming thrombus.

Item Type:Article
Refereed:Yes
Divisions:Life Sciences > School of Biological Sciences > Biomedical Sciences
ID Code:99880
Publisher:MYJoVE Corp

Downloads

Downloads per month over past year

University Staff: Request a correction | Centaur Editors: Update this record

Page navigation