Measurements of Mean Corpuscular Volume and Hemoglobin using Optical Scatter Data from Flow Cytometry

Sammanfattning

Complete blood count (CBC) analysis, often provided by an automated hematology analyzer, is a fundamental diagnostic test for evaluating a patient’s overall health status as a tool in diagnosing various medical conditions. CBC provides insights into the composition of the blood cells with parameters such as Mean Corpuscular Volume (MCV) and Hemoglobin (HGB). MCV represents the average size and volume of red blood cells, while HGB indicates the oxygen-carrying capacity of the blood. Different technologies are used in hematology analyzers, such as the impedance method and spectrophotometry, to achieve precise measurements of MCV and HGB. However, exploring other methodologies is of interest to potentially reduce instrument complexity and cost. Flow cytometry, based on light scatter, provides detailed information on the characteristics of individual cells and is commonly used in CBC analysis to differentiate white blood cells and reticulocytes. However, while the potential of this method for investigating MCV and HGB levels is well established, it is of significant interest to determine if the measurement techniques can be streamlined from three to one by solely using flow cytometry in this prototype analyzer. In this thesis, the possible use of measuring MCV and HGB using a flow cytometry system based on optical scatter data from a prototype hematology analyzer has been examined. The need to sphere the red blood cells before the measurements have also been investigated to evaluate reagent needs. The results have been evaluated based on the correlation factor, accuracy and precision of the proposed optical method. It is shown that the optical method in this thesis, can be used to measure MCV and HGB. However, the necessity of sphering the cells remains. Furthermore, a comparison is made between the optical method and the Sysmex XN-1000 to evaluate the accuracy of the obtained values. Finally, possible improvements and future work are suggested.