Establishment of a flow cytometric method for band 3 detection on reticulocytes: Application to evaluate the band 3 expression between RBCs and reticulocytes in various blood diseases

• Main Authors: Chungming, Chen, 陳俊銘
• Other Authors: 林亮音
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/51932162049945822570

碩士 === 國立臺灣大學 === 醫事技術學研究所 === 91 === Abstract Red blood cells normally have a biconcave discoid shape. In order to maintain this shape, it depends on membrane and cytoskeleton interaction. The red blood cell membrane is a multi-component structure that is responsible for many of the physiological functions and mechanical properties of the cell. Defects in any of these components can manifest as clinical disorders involving the erythrocyte, such as hereditary spherocytosis (HS), hereditary ellipocytosis (HE), or hereditary pyropoikilocytosis (HPP). The first-line clinical laboratory tests for screening red cell membrane disorders, mainly for the diagnosis of HS, include osmotic fragility, the autohaemolysis test, the acid glycerol lysis test, and the pink test. Despite being in use for over 40 years, these laboratory tests have poor specificity and sensitivity due to failure to detect atypical or by factors unrelated to the red cell cytoskeleton defects. In this study, we establish a flow cytometric method to measure the band 3 on reticulocytes. And then, we use this method to measure the fluorescence intensity of mature red blood cells. As a result, the fluorescence intensity of the red blood cells from HS patient was much lower than other anemias. Thus, this EMA method can serve well as a first-line screening test for the diagnosis of hereditary spherocytosis in routine haematology. Besides, only 5μl packed red cells or 10μl whole blood was required, which was suitable for newborn screening. In addition, we found that there should be correlation between cell surface area and band 3 intensity. When we use the method to measure the band 3 intensity on reticulocytes and mature red cells. The ratio is close to 2.0, but lower in leukemia. It suggested that some problems in haematopoiesis in bone marrow might be related with the loss of band 3.