Measuring cell surface area and deformability of individual human red blood cells over blood storage using quantitative phase imaging.

Hyunjoo Park,Yonghak Son,Kyoohyun Kim,Seongsoo Jang,Misuk Ji,Sangyun Lee,Yongkeun Park

doi:10.1038/srep34257

Abstract

The functionality and viability of stored human red blood cells (RBCs) is an important clinical issue in transfusions. To systematically investigate changes in stored whole blood, the hematological properties of individual RBCs were quantified in blood samples stored for various periods with and without a preservation solution called citrate phosphate dextrose adenine-1 (CPDA-1). With 3-D quantitative phase imaging techniques, the optical measurements for 3-D refractive index (RI) distributions and membrane fluctuations were done at the individual cell level. From the optical measurements, the morphological (volume, surface area and sphericity), biochemical (hemoglobin content and concentration), and mechanical parameters (dynamic membrane fluctuation) were simultaneously quantified to investigate the functionalities and progressive alterations of stored RBCs. Our results show that stored RBCs without CPDA-1 had a dramatic morphological transformation from discocytes to spherocytes within two weeks which was accompanied by significant decreases in cell deformability and cell surface area, and increases in sphericity. However, the stored RBCs with CPDA-1 maintained their morphology and deformability for up to 6 weeks.

Highlights

Medicine, University of Ulsan, College of Medicine and Asan Medical Center, Seoul 05535, Republic of Korea. 5TOMOCUBE, Daejeon 34051, Republic of Korea. †Present address: Department of Laboratory Medicine, Konkuk University School of Medicine, Seoul 05030, Republic of Korea. *These authors contributed to this work
red blood cells (RBCs) changes over time in the absence and presence of the preservation solution CPDA-1, 3-D refractive index (RI) tomograms of individual stored RBCs were measured with common-path diffraction optical tomography at days 1, 5, 13, 20, 27, 34 and 41 of the storage period
The RBCs were stored at 4 °C. common-path diffraction optical tomography (cDOT) is a highly sensitive 3-D quantitative phase imaging (QPI) technique which is capable of measuring both the 3-D RI tomogram and dynamic membrane fluctuation of individual cells[23]

Summary

Introduction

It has been reported that progressive deteriorations in the functionalities of RBCs during blood banking/storage were associated with the mortality of transfused patients[15,16,17]. Previous studies on stored RBCs have significantly enhanced our understanding of storage lesions, none of them were able to investigate the morphological, biochemical, and mechanical parameters of individual RBCs simultaneously. The morphological (volume, surface area and sphericity), biochemical (hemoglobin content and concentration), and mechanical parameters (dynamic membrane fluctuation) of individual RBCs were simultaneously quantified to investigate the functionalities and their progressive alterations in stored RBCs. The measurements were done every few days up to 6 weeks to investigate the alterations of these RBC parameters systematically over time in storage. For the stored RBCs without CPDA-1, all the RBCs became spherical shape within a week, whereas the storage agent CPDA-1 prevented the morphological transformation of the RBCs and maintained a healthy level of dynamic membrane fluctuations in the RBCs for up to 6 weeks in storage

Methods

Results

Conclusion