Morphology analysis of unlabeled red blood cells based on quantitative differential phase contrast microscopy

Abstract

The current classical blood smear technique to observe the morphology of single red blood cells (RBCs) for classification is a laborious and error‐prone process. To objectively evaluate the morphology of blood cells, we established a method of computational imaging based on a programmable light emitting diode array. By using quantitative differential phase contrast (qDPC), we characterized the morphology of unlabeled RBCs as well as blood smears. By focusing on comparing the difference of imaging between unlabeled RBCs and stained RBCs under multimode microscopic imaging technology, we demonstrated that qDPC could clearly differentiate discocytes and spherocytes in both unlabeled RBCs and blood smears. The phase map provided by quantitative phase imaging further enhanced the classification accuracy. According to statistical analysis from morphological indexes, the qDPC imaging has a significantly improvement in non‐circularity, texture inhomogeneity and equivalent diameters of cells. Thus, this method has a significant superiority in the capability to analyze the morphology of RBCs and could be applied to clinical assays for determining morphological, functional, and structural deterioration of RBCs.

Quantitative Measurements of Red Blood Cell Indices Using Spectroscopic Differential Phase-Contrast Microscopy

“…The method can also be readily built with inexpensive components such as a programmable LED array and is thus simple and robust. Various forms of DPCs have been demonstrated over the years and have been applied to, for example, measuring sperm cell motility, 49 RBC morphologies, 50 and visualizing birefringence materials. 51 (FPM)-based QPI methods that are aimed to obtain phase images for nondispersive samples at nominally selected wavelengths, 52,53 sDPC employs three color-multiplexed illuminations to obtain three color-dependent phase images, thereby acquiring dispersive information on RBCs.…”

“…The method can also be readily built with inexpensive components such as a programmable LED array and is thus simple and robust. Various forms of DPCs have been demonstrated over the years and have been applied to, for example, measuring sperm cell motility, RBC morphologies, and visualizing birefringence materials . Here, we introduce a functional derivative of DPC microscopy, termed spectroscopic differential phase-contrast (sDPC) microscopy, as a novel detection platform for image-based RBC indices.…”

A Compact Real-time Quantitative Phase Imaging System

“…Compared with other imaging techniques, QPI can quantitatively measure the phase of biological samples with superiority of non-invasive, undamaged and high-definition, so it has become an important optical measurement technology in microscopy. In recent years, QPI techniques based on different principles have been proposed and successfully applied to the study of cell morphology, structure and dynamics [25][26][27][28][29].…”