Innovative Label-Free Lymphoma Diagnosis using Infrared Spectroscopy and Machine Learning on Tissue Sections

The diagnosis of lymphomas is challenging due to their diverse histological presentations and clinical manifestations. There is a need for inexpensive tools that require minimal expertise and are accessible for routine laboratories. Contrastingly, current conventional diagnostic methods are often found only in specialized environments. Attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy offers a nondestructive and user-friendly approach in the analysis of a wide range of samples. In this paper, we determined whether the technique coupled with machine learning can detect and differentiate lymphoma within lymphoid tissue samples. Tissue sections from 295 individuals diagnosed with lymphoma and 389 individuals without the disease were analyzed using ATR-FTIR spectroscopy. The resulting spectral dataset was split using a 70:30 train-test split. Partial least Squares Discriminant Analysis (PLS-DA) models were trained to distinguish non-malignant lymphoid tissue from lymphoma samples and to differentiate between subtypes. On the training set (n=478), significant spectral differences were mainly identified in the 1800-900 cm-1 region, attributed to fundamental biochemical constituents like proteins, lipids, carbohydrates, and nucleic acids. On the independent test set (n=206), the trained PLS-DA model achieved a promising AUC of 0.882 (95% CI: 0.881-0.884) in the differentiation between lymphoma and non-malignant lymphoid tissue. In addition, comparative analyses revealed spectral distinctions and notable clustering between the different lymphoma subtypes. This study provides valuable insights into the application of ATR-FTIR spectroscopy and machine learning in the field of lymphoma diagnosis as a novel, non-destructive, rapid and inexpensive tool with the potential to be easily implemented in non-specialized laboratories.