Skip to main content

Assessment of Platelet Function by High-Throughput Screening Light Transmission Aggregometry: Optimul Assay

  • Protocol
  • First Online:
Hemostasis and Thrombosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2663))

Abstract

Platelet function testing is critical in the diagnosis of bleeding disorders and allows monitoring of antiplatelet therapy. The gold standard assay, light transmission aggregometry (LTA), was developed 60 years ago and remains widely used worldwide. It requires, however, access to expensive equipment and is time-consuming, and the interpretation of results requires evaluation by an experienced investigator. It also suffers from a lack of standardization, resulting in widely variable results between laboratories. 96-well plate-based Optimul aggregometry utilizes the same principles of LTA and aims to standardize agonist concentrations with the development of 96-well plates which are precoated with 7 concentrations of each lyophilized agonist (arachidonic acid, adenosine diphosphate, collagen, epinephrine, TRAP-6 amide, and U46619) and stored at ambient room temperature (20–25 °C) for up to 12 weeks. For platelet function testing, 40 μL of platelet-rich plasma is added to each well, and the plate is placed onto a plate shaker, after which platelet aggregation is determined by changes in light absorbance. This method reduces the blood volume required and allows for in-depth platelet function analysis without specialist training, or the need to purchase expensive, dedicated equipment.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 219.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 279.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Hvas A-M, Favaloro EJ (2017) Platelet function analyzed by light transmission aggregometry. Methods Mol Biol 1646:321–331. https://doi.org/10.1007/978-1-4939-7196-1_25

    Article  CAS  PubMed  Google Scholar 

  2. Michelson AD (2004) Platelet function testing in cardiovascular diseases. Circulation 110:e489–e493. https://doi.org/10.1161/01.CIR.0000147228.29325.F9

    Article  PubMed  Google Scholar 

  3. Cattaneo M, Hayward CPM, Moffat KA et al (2009) Results of a worldwide survey on the assessment of platelet function by light transmission aggregometry: a report from the platelet physiology subcommittee of the SSC of the ISTH. J Thromb Haemost 7:1029–1029. https://doi.org/10.1111/j.1538-7836.2009.03458.x

    Article  CAS  PubMed  Google Scholar 

  4. Cattaneo M, Cerletti C, Harrison P et al (2013) Recommendations for the standardization of light transmission aggregometry: a consensus of the working party from the platelet physiology subcommittee of SSC/ISTH. J Thromb Haemost 11:1183–1189. https://doi.org/10.1111/jth.12231

    Article  Google Scholar 

  5. Dawood BB, Lowe GC, Lordkipanidzé M et al (2012) Evaluation of participants with suspected heritable platelet function disorders including recommendation and validation of a streamlined agonist panel. Blood 120:5041–5049. https://doi.org/10.1182/blood-2012-07-444281

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Gresele P, Subcommittee on Platelet Physiology of the International Society on Thrombosis and Hemostasis (2015) Diagnosis of inherited platelet function disorders: guidance from the SSC of the ISTH. J Thromb Haemost 13:314–322. https://doi.org/10.1111/jth.12792

    Article  CAS  PubMed  Google Scholar 

  7. Chan MV, Armstrong PC, Warner TD (2018a) 96-well plate-based aggregometry. Platelets 29:650–655. https://doi.org/10.1080/09537104.2018.1445838

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Fratantoni JC, Poindexter BJ (1990) Measuring platelet aggregation with microplate reader. A new technical approach to platelet aggregation studies. Am J Clin Pathol 94:613–617

    Article  CAS  PubMed  Google Scholar 

  9. Chan MV, Leadbeater PD, Watson SP, Warner TD (2018b) Not all light transmission aggregation assays are created equal: qualitative differences between light transmission and 96-well plate aggregometry. Platelets 29:686–689. https://doi.org/10.1080/09537104.2018.1466388

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Chan MV, Armstrong PCJ, Papalia F et al (2011) Optical multichannel (optimul) platelet aggregometry in 96-well plates as an additional method of platelet reactivity testing. Platelets 22:485–494. https://doi.org/10.3109/09537104.2011.592958

    Article  CAS  PubMed  Google Scholar 

  11. Chan MV, Warner TD (2012) Standardised optical multichannel (optimul) platelet aggregometry using high-speed shaking and fixed time point readings. Platelets 23:404–408. https://doi.org/10.3109/09537104.2011.603066

    Article  CAS  PubMed  Google Scholar 

  12. Lordkipanidzé M, Lowe GC, Kirkby NS et al (2014) Characterization of multiple platelet activation pathways in patients with bleeding as a high-throughput screening option: use of 96-well Optimul assay. Blood 123:e11–e22. https://doi.org/10.1182/blood-2013-08-520387

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Paterson GG, Young JM, Willson JA et al (2020) Hypoxia modulates platelet purinergic signalling pathways. Thromb Haemost 120:253–261. https://doi.org/10.1055/s-0039-3400305

    Article  PubMed  Google Scholar 

  14. Hsu H, Chan MV, Armstrong PC et al (2022) A pilot study assessing the implementation of 96 well plate-based aggregometry (Optimul) in Australia. Pathology 54(6):746–754. https://doi.org/ 10.1016/j.pathol.2022.03.012

    CAS  Google Scholar 

  15. Grech J, Chan MV, Ochin C et al (2022) Serotonin-affecting antidepressant use in relation to platelet reactivity. Clin Pharmacol Ther 111:909–918. https://doi.org/10.1002/cpt.2517

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Harrison P, Mackie I, Mumford A et al (2011) Guidelines for the laboratory investigation of heritable disorders of platelet function. Br J Haematol 155:30–44. https://doi.org/10.1111/j.1365-2141.2011.08793.x

    Article  CAS  PubMed  Google Scholar 

  17. R Core Team (2021) R: A language and environment for statistical computing

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Melissa V. Chan .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Chan, M.V., Lordkipanidzé, M., Warner, T.D. (2023). Assessment of Platelet Function by High-Throughput Screening Light Transmission Aggregometry: Optimul Assay. In: Favaloro, E.J., Gosselin, R.C. (eds) Hemostasis and Thrombosis. Methods in Molecular Biology, vol 2663. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3175-1_41

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3175-1_41

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3174-4

  • Online ISBN: 978-1-0716-3175-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics