Flow morphometry to assess the red blood cell storage lesion

Abstract

We present a novel automated system for morphology analysis of red blood cells (RBC) under flow. RBC concentrates collected by blood banks for transfusions are stored for periods of up to several weeks, during which time a number of changes occur, collectively termed the storage lesion. Typically the extent of hemolysis is the defining criterion to determine the acceptability of the RBCs for transfusions. Morphological changes are related with biochemical alteration during the storage of RBCs. The typical blood smear procedure for determining such changes is a labor-intensive and potentially biased manual process. The advantage of the flow morphometry system presented here is that it provides fully automated morphological classification of RBCs with large sample numbers in a short time. Our system uses a commercially available flow cell and flow conditions that prevent adhesion of RBCs, thus eliminating the need for blocking agents such as albumin that affect the distribution of cell shapes. Our morphometry results are validated by comparison with standard biochemical assays (hemolysis, ATP) for blood from 17 donors stored under blood bank conditions for 13 weeks. We show that the percentage of spherocytes present can be used to estimate the status of RBC concentrates. © 2017 International Society for Advancement of Cytometry.

ACVIM consensus statement on the diagnosis of immune‐mediated hemolytic anemia in dogs and cats

“…Spherocytes should be used as a diagnostic criterion only in dogs because feline erythrocytes do not consistently display central pallor. Interpreting spherocytes after blood transfusion should be done cautiously, because stored blood products may contain high proportions of spherocytes and spherocytes have been documented in human patients with hemolytic transfusion reactions . Spherocytosis should be assessed in the monolayer of a well‐made blood smear, because spherocyte‐like artifacts arise toward the feathered edge and in thick areas .…”

Label‐Free Analysis of Red Blood Cell Storage Lesions Using Imaging Flow Cytometry

“…Three-dimensional refractive index tomograms (37) and digital holographic microscopy images (38) have been used to measure temporal changes in morphological parameters of stored RBCs (39,40). Flow morphometry techniques, such as the fluidic system by Sierra et al, acquire high-throughput LM images of RBCs in flow suspension (41). In their work, a threshold-based binary decision tree was applied for the automated classification of RBCs into discocytes, echinocytes, and spherocytes.…”

Hemolysis Pathways during Storage of Erythrocytes and Inter-Donor Variability in Erythrocyte Morphology

“…This does, however, lead to the limitation that the high and low hemolysis samples are only identified at the end of the storage period, so that we are not observing the change in the RBC morphology over the course of the storage period. Previous work in which the morphology was measured as a function of time for a small number of samples and compared to the hemolysis does not appear to offer additional information [23].…”

“…It is, however, important to differentiate between measuring changes in a single sample or a defined group of samples, and the use of the MI as a marker of storage quality for an individual measurement. The inter-donor variability in morphology that has been noted here would not affect the morphological analysis used to assess the response to toxins [26], the effects of sample transportation [27], variation of samples with time [23], or the comparison of the effects of storage media [28].…”