High-frequency photoacoustic microscopy of hematoxylin and eosin stained slides for veterinary oncology applications

We tested high-frequency photoacoustic imaging for efficacy in recovering the histopathological structures from hematoxylin and eosin (H&E) stained tissue sections. The results showed that exogenous staining improved the absorption contrast of the histology microscope slides, allowing the detection of adipose tissue structure with a micrometer resolution throughout the tens of micrometer-square area range. The comparative spectroscopic study proved that photoacoustic imaging caused two simultaneously occurring processes: photodegradation of H&E (and formation of stable photoproducts absorbing in the blue spectral region), and ultrasound assisted organic stain removal from the microscope slide sample, reducing the exogenous contrast-distinguishable photoacoustic detection area.