Polarized Hyperspectral Microscopic Imaging for White Blood Cells on Wright-Stained Blood Smear Slides.

White blood cells, also called leukocytes, are hematopoietic cells of the immune system that are involved in protecting the body against both infectious diseases and foreign materials. The abnormal development and uncontrolled proliferation of these cells can lead to devastating cancers. Their timely recognition in the peripheral blood is critical to diagnosis and treatment. In this study, we developed a microscopic imaging system for improving the visualization of white blood cells on Wright's stained blood smear slides, with two different setups: polarized light imaging and polarized hyperspectral imaging. Based on the polarized light imaging setup, we collected the RGB images of Stokes vector parameters (S0, S1, S2, and S3) of five types of white blood cells (neutrophil, eosinophil, basophil, lymphocyte, and monocyte), and calculated the Stokes vector derived parameters: the degree of polarization (DOP), the degree of linear polarization (DOLP), and the degree of circular polarization (DOCP)). We also calculated Stokes vector data based on the polarized hyperspectral imaging setup. The preliminary results demonstrate that Stokes vector derived parameters (DOP, DOLP, and DOCP) could improve the visualization of granules in granulocytes (neutrophils, eosinophils, and basophils). Furthermore, Stokes vector derived parameters (DOP, DOLP, and DOCP) could improve the visualization of surface structures (protein patterns) of lymphocytes enabling subclassification of lymphocyte subpopulations. Finally, S2, S3, and DOCP could enhance the morphologic visualization of monocyte nucleus. We also demonstrated that the polarized hyperspectral imaging setup could provide complementary spectral information to the spatial information on different Stokes vector parameters of white blood cells. This work demonstrates that polarized light imaging & polarized hyperspectral imaging has the potential to become a strong imaging tool in the diagnosis of disorders arising from white blood cells.

A Method to Determine Human Skin Heat Capacity Using a Non-Invasive Calorimetric Sensor.

A calorimetric sensor has been designed to measure the heat flow dissipated by a 2 x 2 cm2 skin surface. In this work, a non-invasive method is proposed to determine the heat capacity and thermal conductance of the area of skin where the measurement is made. The method consists of programming a linear variation of the temperature of the sensor thermostat during its application to the skin. The sensor is modelled as a two-inputs and two-outputs system. The inputs are 1) the power dissipated by the skin and transmitted by conduction to the sensor, and 2) the power dissipated in the sensor thermostat to maintain the programmed temperature. The outputs are 1) the calorimetric signal and 2) the thermostat temperature. The proposed method consists of a sensor modelling that allows the heat capacity of the element where dissipation takes place (the skin) to be identified, and the transfer functions (TF) that link the inputs and outputs are constructed from its value. These TFs allow the determination of the heat flow dissipated by the surface of the human body as a function of the temperature of the sensor thermostat. Furthermore, as this variation in heat flow is linear, we define and determine an equivalent thermal resistance of the skin in the measured area. The method is validated with a simulation and with experimental measurements on the surface of the human body.

Hyperspectral imaging for head and neck cancer detection: specular glare and variance of the tumor margin in surgical specimens.

Head and neck squamous cell carcinoma (SCC) is primarily managed by surgical cancer resection. Recurrence rates after surgery can be as high as 55%, if residual cancer is present. Hyperspectral imaging (HSI) is evaluated for detection of SCC in ex-vivo surgical specimens. Several machine learning methods are investigated, including convolutional neural networks (CNNs) and a spectral-spatial classification framework based on support vector machines. Quantitative results demonstrate that additional data preprocessing and unsupervised segmentation can improve CNN results to achieve optimal performance. The methods are trained in two paradigms, with and without specular glare. Classifying regions that include specular glare degrade the overall results, but the combination of the CNN probability maps and unsupervised segmentation using a majority voting method produces an area under the curve value of 0.81 [0.80, 0.83]. As the wavelengths of light used in HSI can penetrate different depths into biological tissue, cancer margins may change with depth and create uncertainty in the ground truth. Through serial histological sectioning, the variance in the cancer margin with depth is investigated and paired with qualitative classification heat maps using the methods proposed for the testing group of SCC patients. The results determined that the validity of the top section alone as the ground truth may be limited to 1 to 2 mm. The study of specular glare and margin variation provided better understanding of the potential of HSI for the use in the operating room.

Cancer Detection Using Hyperspectral Imaging and Evaluation of the Superficial Tumor Margin Variance with Depth.

Head and neck squamous cell carcinoma (SCCa) is primarily managed by surgical resection. Recurrence rates after surgery can be as high as 55% if residual cancer is present. In this study, hyperspectral imaging (HSI) is evaluated for detection of SCCa in ex-vivo surgical specimens. Several methods are investigated, including convolutional neural networks (CNNs) and a spectral-spatial variant of support vector machines. Quantitative results demonstrate that additional processing and unsupervised filtering can improve CNN results to achieve optimal performance. Classifying regions that include specular glare, the average AUC is increased from 0.73 [0.71, 0.75 (95% confidence interval)] to 0.81 [0.80, 0.83] through an unsupervised filtering and majority voting method described. The wavelengths of light used in HSI can penetrate different depths into biological tissue, while the cancer margin may change with depth and create uncertainty in the ground-truth. Through serial histological sectioning, the variance in cancer-margin with depth is also investigated and paired with qualitative classification heat maps using the methods proposed for the testing group SCC patients.