High-content stimulated Raman histology of human breast cancer

Histological examination is crucial for cancer diagnosis, including hematoxylin and eosin (H&E) staining for mapping morphology and immunohistochemistry (IHC) staining for revealing chemical information. Recently developed two-color stimulated Raman histology could bypass the complex tissue processing to mimic H&E-like morphology. Yet, the underlying chemical features are not revealed, compromising the effectiveness of prognostic stratification. Here, we present a high-content stimulated Raman histology (HC-SRH) platform that provides both morphological and chemical information for cancer diagnosis based on un-stained breast tissues. Through spectral unmixing in the C-H vibration window, HC-SRH can map unsaturated lipids, cellular protein, extracellular matrix, saturated lipid, and water in breast tissue. In this way, HC-SRH provides excellent contrast for various tissue components. Considering rapidness is important in clinical trials, we implemented spectral selective sampling to boost the speed of HC-SRH by one order. We also successfully demonstrated the HC-SRH in a clinical-compatible fiber laser-based SRS microscopy. With the widely rapid tuning capability of the advanced fiber laser, a clear chemical contrast of nucleic acid and solid-state ester is shown in the fingerprint result.