Current Landscape of Advanced Imaging Tools for Pathology Diagnostics

Abstract

Histopathology relies on century-old workflows of formalin fixation, paraffin embedding, sectioning, and staining tissue specimens on glass slides. Despite being robust, this conventional process is slow, labor-intensive, and limited to providing two-dimensional views. Emerging technologies promise to enhance and accelerate histopathology. Slide-free microscopy allows rapid imaging of fresh, unsectioned specimens, overcoming slide preparation delays. Methods such as fluorescence confocal microscopy, multiphoton microscopy, along with more recent innovations including microscopy with UV surface excitation and fluorescence-imitating brightfield imaging can generate images resembling conventional histology directly from the surface of tissue specimens. Slide-free microscopy enable applications such as rapid intraoperative margin assessment and, with appropriate technology, three-dimensional histopathology. Multiomics profiling techniques, including imaging mass spectrometry and Raman spectroscopy, provide highly multiplexed molecular maps of tissues, although clinical translation remains challenging. Artificial intelligence is aiding the adoption of new imaging modalities via virtual staining, which converts methods such as slide-free microscopy into synthetic brightfield-like or even molecularly informed images. Although not yet commonplace, these emerging technologies collectively demonstrate the potential to modernize histopathology. Artificial intelligence-assisted workflows will ease the transition to new imaging modalities. With further validation, these advances may transform the century-old conventional histopathology pipeline to better serve 21st-century medicine. This review provides an overview of these enabling technology platforms and discusses their potential impact.

Keywords: artificial intelligence; histology; microscopy; multiomics; slide-free; stain-free.