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Applications of imaging flow cytometry in the diagnostic assessment of red cell membrane disorders.
Cytometry Part B: Clinical Cytometry ( IF 2.3 ) Pub Date : 2019-11-21 , DOI: 10.1002/cyto.b.21857
Tejashree Anil More 1 , Bhavik Dalal 1 , Rati Devendra 1 , Prashant Warang 1 , Aruna Shankarkumar 1 , Prabhakar Kedar 1
Affiliation  

BACKGROUND Red cell membranopathies refers to phenotypically and morphologically heterogeneous disorders. High throughput imaging flow cytometry (IFC) combines the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy to produce high content image analysis with quantitative analysis. We have evaluated the applications of IFC to examine both the morphology as well as fluorescence signal intensity in red cell membranopathies. METHODS Fluorescence intensity of eosin-5-maleimide (EMA) labeled red cells was measured for diagnosis of RBC membrane protein defect on Amnis ImageStreamX followed by Image analysis on IDEAS software to study features such as circularity and shape ratio. RESULTS The hereditary spherocytosis (HS) group showed significantly decreased MFI (52,800 ± 9,100) than normal controls (81,100 ± 4,700) (p < .05) whereas non-HS showed 78,300 ± 9,900. The shape ratio of hereditary elliptocytosis (HE) was significantly higher (43.8%) than normal controls (14.6%). The circularity score is higher in HS (64.15%) than the normal controls (44.3%) whereas the circularity score was very less in HE (10%) due to the presence of elliptocytes. CONCLUSIONS The advantages of the IFC over standard flow cytometry is its ability to provide high-content image analysis and measurement of parameters such as circularity and shape ratio allow discriminating red cell membranopathies (HS and HE) due to variations in shape and size. It could be a single, effective, and rapid IFC test for detection and differentiation of red cell membrane disorders in hematology laboratories where an IFC is available.

中文翻译:

成像流式细胞术在红细胞膜疾病诊断评估中的应用。

背景技术红细胞膜病是指表型和形态上的异质性疾病。高通量成像流式细胞术(IFC)将流式细胞术的速度,灵敏度和表型能力与显微镜的详细图像和功能见解相结合,可进行定量分析的高含量图像分析。我们评估了IFC在检查红细胞膜病中的形态以及荧光信号强度方面的应用。方法在Amnis ImageStreamX上测量曙红5-马来酰亚胺(EMA)标记的红细胞的荧光强度,以诊断RBC膜蛋白缺陷,然后在IDEAS软件上进行图像分析,以研究圆度和形状比等特征。结果遗传性球菌增多症(HS)组的MFI显着降低(52,800±9,100)比正常对照组(81,100±4,700)(p <.05),而非HS则为78,300±9,900。遗传性椭圆细胞增多症(HE)的形状比(43.8%)显着高于正常对照组(14.6%)。HS的圆形度得分(64.15%)比正常对照组(44.3%)高,而HE的圆形度得分(10%)由于椭圆细胞的存在而非常低。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析能力,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜细胞病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。100±4,700)(p <.05),而非HS则显示78,300±9,900。遗传性椭圆细胞增多症(HE)的形状比(43.8%)显着高于正常对照组(14.6%)。HS的圆形度得分(64.15%)比正常对照组(44.3%)高,而HE的圆形度得分(10%)由于椭圆细胞的存在而非常低。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析能力,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜细胞病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。100±4,700)(p <.05),而非HS则显示78,300±9,900。遗传性椭圆细胞增多症(HE)的形状比(43.8%)显着高于正常对照组(14.6%)。HS的圆形度得分(64.15%)比正常对照组(44.3%)高,而HE的圆形度得分(10%)由于椭圆细胞的存在而非常低。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析能力,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜细胞病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。900.遗传性椭圆细胞增多症(HE)的形状比(43.8%)显着高于正常对照组(14.6%)。HS的圆形度得分(64.15%)比正常对照组(44.3%)高,而HE的圆形度得分(10%)由于椭圆细胞的存在而非常低。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析能力,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜细胞病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。900.遗传性椭圆细胞增多症(HE)的形状比(43.8%)显着高于正常对照组(14.6%)。HS的圆形度得分(64.15%)比正常对照组(44.3%)高,而HE的圆形度得分(10%)由于椭圆细胞的存在而非常低。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析功能,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜性病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。6%)。HS的圆形度得分(64.15%)比正常对照组(44.3%)高,而HE的圆形度得分(10%)由于椭圆细胞的存在而非常低。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析能力,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜细胞病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。6%)。HS的圆形度得分(64.15%)比正常对照组(44.3%)高,而HE的圆形度得分(10%)由于椭圆细胞的存在而非常低。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析能力,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜细胞病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析功能,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜性病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。结论与标准流式细胞术相比,IFC的优势在于它能够提供高含量的图像分析能力,并且能够测量诸如圆形度和形状比之类的参数,从而能够区分由于形状和大小变化而引起的红细胞膜细胞病变(HS和HE)。它可以是在有IFC的血液学实验室中检测和区分红细胞膜疾病的单一,有效且快速的IFC测试。
更新日期:2019-11-21
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