Hyperspectral imaging for skin cancer and blood disorders diagnosis

Hyperspectral imaging is a novel technology for acquiring an image at a large number of wavelengths, thus allowing the study of spectral and spatial details of a sample under analysis. This technology has emerged as a promising imaging modality to be used as a diagnostic tool in several medical applications where spectral information is relevant. In this paper, we outline our most recent achievements in this field. Firstly, hyperspectral imaging systems developed to improve non-invasive diagnosis of skin cancer, consisting of digital silicon and InGaAs cameras and light emitting diodes, are described. Secondly, we present our last advances using hyperspectral technology together with confocal microscopy to improve the diagnosis of blood diseases, in particular, hemoglobinopathies such as thalassemia and cell membrane diseases such as hereditary spherocytosis. Finally, new insights on these topics are discussed.This project has been funded by the Agencia Estatal de Investigación (AEI) (PID2020-112527RB-I00 / AEI / 10.13039/501100011033). L R-B thanks the Ministry of Science, Innovation and Universities for the PhD (FPI) grant she has received.Peer ReviewedPostprint (author's final draft

Membrane protein detection and morphological analysis of red blood cells in hereditary spherocytosis by confocal laser scanning microscopy

In hereditary spherocytosis (HS), genetic mutations in the cell membrane and cytoskeleton proteins cause structural defects in red blood cells (RBCs). As a result, cells are rigid and misshapen, usually with a characteristic spherical form (spherocytes), too stiff to circulate through microcirculation regions, so they are prone to undergo hemolysis and phagocytosis by splenic macrophages. Mild to severe anemia arises in HS, and other derived symptoms like splenomegaly, jaundice, and cholelithiasis. Although abnormally shaped RBCs can be identified under conventional light microscopy, HS diagnosis relies on several clinical factors and sometimes on the results of complex molecular testing. It is specially challenging when other causes of anemia coexist or after recent blood transfusions. We propose two different approaches to characterize RBCs in HS: (i) an immunofluorescence assay targeting protein band 3, which is affected in most HS cases and (ii) a three-dimensional morphology assay, with living cells, staining the membrane with fluorescent dyes. Confocal laser scanning microscopy (CLSM) was used to carry out both assays, and in order to complement the latter, a software was developed for the automated detection of spherocytes in blood samples. CLSM allowed the precise and unambiguous assessment of cell shape and protein expression.This publication is part of the project PID2020-112527RBI00, funded by CIN/AEI/10.13039/501100011033. Laura Rey-Barroso thanks the Ministry of Science, Innovation and Universities for the PhD (FPI) grant she has received (DPI2017-89414-R). The current study has been funded by the Spanish National Agency of Investigation (AEI).Peer ReviewedPostprint (published version