Polarized hyperspectral and polarized light microscopic imaging for enhanced visualization of white blood cells

White blood cells (WBC) are hematopoietic cells of the immune system that protect the body by recognizing and eliminating infectious agents. Abnormalities in WBC production, maturation, or function can lead to disease and associated morphologic changes that, when systematically characterized, support diagnostic classification and clinical decision-making.

We aim to investigate polarized hyperspectral imaging (PHSI) and polarized light imaging (PLI) microscopy for the visualization of WBCs.

We developed a dual-modality microscopic imaging system that performs both polarized hyperspectral imaging and polarized light imaging. In the dual imaging setup, we used a snapscan hyperspectral camera and an RGB camera to acquire images separately and further calculate four Stokes parameters (S0, S1, S2, and S3) as well as three Stokes vector-derived parameters, namely, the degree of polarization, degree of linear polarization, and degree of circular polarization. Synthetic RGB images of Stokes vectors and Stokes vector-derived parameters were generated for the visualization of cellular components with PHSI images. The spectral signatures of representative WBCs, e.g., granulocytes and lymphocytes, were extracted for qualitative comparison.

The preliminary results demonstrate that Stokes vector parameters can enhance the visualization of granules in granulocytes, the visualization of surface structures of lymphocytes, and the morphologic visualization of the monocyte nucleus. Furthermore, the results also reveal that the measured spectra of Stokes vector parameters could enhance the differentiation of WBCs in the spectral dimension, represented by the qualitative comparison between granulocytes and lymphocytes.

Utilizing the spatial and spectral information from the Stokes vector data, our customized polarized hyperspectral microscopic imaging system enhances the visualization of WBCs and may provide a tool for the diagnosis of disorders related to white blood cells.