Mass cytometry of platelet-rich plasma: a new approach to analyze platelet surface expression and reactivity

Platelet Subtypes in Inflammatory Settings

“…The way to solve the inherited limitation of FFC is to overcome the spectral overlap and have the possibility to evaluate different markers simultaneously on individual platelets. One way to achieve that is by applying mass cytometry (MC), which is a next-generation flow cytometry platform and using probes that are conjugated to heavy metal isotopes instead of fluorescent dyes and time-of-flight as a detection technique ( 78 , 79 ). Using MC there will be no need for compensation as this detection technique has minimal spectral or channel overlap resulting in an increase in the number of cellular parameters that can be analyzed simultaneously on individual cells.…”

“…One of the limitations with platelet subsets studies, in general, is the lack of standardized protocols that are easily reproducible. Recently, a structured method to stain and evaluate platelets from PRP using CyTOF was published which allows the acquisition of 300,000 to 500,000 events and recording the expression of up to 40 markers at once ( 78 ). MC data can be analyzed using Visual stochastic neighbor embedding (viSNE) to visualize high-dimensional single-cell data, for platelet-specific analysis some groups have developed freely available analysis pipelines such as CYANUS ( 83 ).…”

Preanalytical conditions for multiparameter platelet flow cytometry

“…Accordingly, there has been a parallel expansion and development of multidimensional analytical tools such as t-stochastic neighborhood embedded (tSNE) [ 3 ], spanning-tree progression analysis of density-normalized events (SPADE) [ 4 ], and flow self-organizing maps (flowSOM) [ 5 ]. Concurrent with advances in the wider field of single-cell analysis, platelet flow cytometry has seen various innovations including multidimensional FFC analysis [ [6] , [7] , [8] ], phosphoflow and barcoding [ 9 ], application of spectral FFC [ 10 ] and CyTOF with multidimensional analysis [ [11] , [12] , [13] ]. While the latter may show comparable efficacy to FFC [ 14 ] and the design of CyTOF assays has been recently described in full [ 15 ], it remains relatively inaccessible when compared with FFC, which is common in research and medical institutes worldwide.…”