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The study of platelet aggregation using a microtiter plate reader ‒ methodological considerations
Magdalena Boncler1, Jacek Golański2
1Department of Haemostasis and Haemostatic Disorders, Chair of Biomedical Sciences, Medical University of Łódź, ul. Mazowiecka 6/8, Łódź, 92-215, Poland. magdalena.boncler@umed.lodz.pl.
Abstract:
Optical aggregometry by 96-well plate assay, the microplate method, is a fast, efficient, and readily available method for measuring the pharmacological effects of antiplatelet drugs. Even though recent years have witnessed growing interest in adopting the microplate method for widespread use, it remains in the shadow of the standard light transmission aggregometry (LTA). Regardless of the method used, the results of platelet aggregation depend on a variety of factors and often vary among laboratories worldwide. While several methodological papers have examined the microplate method, no standards have been established, most likely because the approach is not used as a diagnostic tool. Currently, the microplate method is recommended by researchers to be used in conjunction with LTA or as an adjunct to LTA. This raises the question of whether an optimal protocol exists for microplate aggregometry, and what are the key considerations in a good experimental protocol for obtaining reliable results? This article attempts to address these questions by summarizing the knowledge accumulated in this field over the last three decades.
Insights
Microplate aggregometry offers a fast, efficient method for studying antiplatelet drugs, though it requires standardized protocols for reliable results. This review explores optimal methods for this technique, comparing it to light transmission aggregometry.
Area of Science:
- Pharmacology
- Hematology
- Analytical Chemistry
Background:
- Optical aggregometry using 96-well plates (microplate method) is a rapid and accessible technique for assessing antiplatelet drug efficacy.
- Despite its advantages, the microplate method is less utilized than standard light transmission aggregometry (LTA).
- Platelet aggregation results are influenced by numerous factors, leading to inter-laboratory variability, and standardized protocols for the microplate method are lacking.
Purpose of the Study:
- To investigate the existence of an optimal protocol for microplate aggregometry.
- To identify key considerations for developing a reliable experimental protocol for microplate aggregometry.
- To summarize existing knowledge on microplate aggregometry over the past three decades.
Main Methods:
- Review of methodological papers and accumulated knowledge on microplate aggregometry over 30 years.
- Comparison of microplate method with standard light transmission aggregometry (LTA).
- Discussion of factors influencing platelet aggregation results and the need for standardization.
Main Results:
- The microplate method is recognized for its speed and efficiency in evaluating antiplatelet agents.
- Current recommendations suggest using the microplate method in conjunction with or as an adjunct to LTA.
- Lack of established standards hinders widespread adoption, particularly for diagnostic purposes.
Conclusions:
- An optimal protocol for microplate aggregometry is yet to be universally established.
- Further research and standardization are crucial for the reliable application of microplate aggregometry.
- The microplate method holds potential as a valuable tool when used with appropriate methodological considerations.
