Research On The Classification And Identification Of Whole Blood Cells Based On Flow Cytometry And Quantitative Phase Microscopy

The medical test results of human blood cells have become important information for diagnosing the health of patients.The working principle of common blood cell analyzers is often combined with three-dimensional fluid focusing technology,electrical impedance analysis technology,cell fluorescence specific microscopy,and cell light scattering signal analysis.The blood cell analyzer can present statistical results of its data,including the absolute number,relative proportion,and hemoglobin of various types of cells per unit volume.Similarly,quantitative phase microscopy has made great progress and development in biological cell imaging in recent years.Different refractive index profiles and cell optical thicknesses cause different phase shifts of light.Therefore,quantitative phase microscopy is suitable for imaging blood cells.Quantitative phase imaging plays an important role in the acquisition of internal substructure information of unknown cells,the measurement of relative refractive index of cells,and the detection of cancerous cells.Quantitative phase imaging enables precise detection of cells without destroying the biological activity of the cells and without changing the physical appearance of the cells.In the traditional flow cytometry analysis,the cell substructure morphological recognition process often only uses the forward and side scatter light signals of the cells as the basis for classification.Also,fluorescent staining techniques alter cell viability.Firstly,this paper proposes a method combining the forward and backward scattered light intensity information of cells as a method for classifying and identifying white blood cell subclasses.Secondly,based on the actual lymphocyte and eosinophil morphological characteristics,we established the corresponding monocyte model and binuclear cell model.The Mie scattering formula and the Rayleigh-Debye-Gans approximation method under spherical particles are introduced.A simulated experimental optical path capable of simultaneously receiving forward and backward scatter spectra of cells was designed using VirtualLab Fusion software.Simulation experiments show the scattering profiles of the two cell models.Next,we established the relationship between the anterior and posterior scattered light intensity of the cells at different relative refractive indices and the incident wavelengths of 442 nm,532 nm and 633 nm.Further,it is found that lymphocytes and eosinophils are significantly different under the characterization that the backscattered light signal is the vertical axis and the forward scattered light signal is the horizontal axis.Finally,a classification method for lymphocytes and eosinophils was proposed,and the effectiveness of the method was demonstrated from the classification results.Not only is leukocyte subclass analysis,but morphological analysis and signature recognition of whole blood cells(leukocytes,red blood cells,and platelets)are also important in life sciences and clinical medicine.Based on the principles of Diffraction Quantitative Phase Imaging and Light Intensity Transfer Equation(TIE),we performed experiments using the Biophase Instrument.Optical phase information of red blood cells,platelets,and neutrophils was obtained.In order to quantitatively identify different types of blood cells,five characteristic parameters of cell phase measurement are introduced,including phase peak-to-valley difference,phase volume,phase surface area,phase projection perimeter,and phase projection area.At the same time,the corresponding phase characteristic values of red blood cells,neutrophils and platelets were discussed.In addition,a phase gradient method is proposed for the "halo"(phase delay)phenomenon that often occurs around cells during phase acquisition.The purpose is to remove the halo around the object to be measured,resulting in a clearer cell boundary profile.Finally,combined with the threshold and phase gradient calculation of cell phase characteristic parameters,a classification and recognition method for identifying the above three types of human blood cells was established.The experimental results show that the method has good accuracy and feasibility.This provides an idea for the classification of blood cell analysis based on quantitative phase imaging.