The use of intracellular dyes to create a multiplexed flow cytometry-based red blood cell phenotyping assay.

Background And Objectives: Flow cytometry can be used to phenotype red blood cell antigens, allowing for high-throughput testing while using low reagent volumes. This article utilizes intracellular dyes to pre-label red blood cells to further multiplex flow cytometry-based red blood cell antigen phenotyping.

Materials And Methods: Red blood cells were pre-labelled using the intracellular dyes V450 and Oregon Green.

Development of multiplexed flow cytometry-based red blood cell antibody screen and identification assays.

Background And Objectives: The purpose of this study was to develop a high-throughput method of performing red blood cell antibody screens and identification by utilizing flow cytometry and intracellular dyes to allow a multiplexed assay where three-cell screens can be performed in a single test well and 11-cell panels in three test wells.

Materials And Methods: Reagent red blood cells were labelled using Violet Proliferation Dye 450 (V450) and Oregon Green fluorescent dyes, which bind intracellular proteins to allow up to four cells to be interrogated in a single test well. Sixteen 3-cell screen panels and ten 11-cell identification panels were tested using sera with known antibody specificity.

Validation of a flow-cytometry-based red blood cell antigen phenotyping method.

Background And Objectives: Current manual and automated phenotyping methods are based on visual detection of the antigen-antibody interaction. This approach has several limitations including the use of large volumes of patient and reagent red blood cells (RBCs) and antisera to produce a visually detectable reaction. We sought to determine whether the flow cytometry could be developed and validated to perform RBC phenotyping to enable a high-throughput method of phenotyping using comparatively miniscule reagent volumes via fluorescence-based detection of antibody binding.

Implementation of a novel real-time platelet inventory management system at a multi-site transfusion service.

Background: Blood platelets (PLTs) are a valuable commodity. Management of their inventory has implications both for patient care and for the cost of health care delivery. There are a variety of different methods of managing PLT inventory currently in practice and multiple theoretical models aimed at improving PLT inventory metrics.