Digital pathology using whole slide images has been recently approved for primary diagnosis in clinical surgical pathology practices. Here we describe a novel imaging method, fluorescence-imitating brightfield imaging, that can capture the surface of fresh tissue without requiring prior fixation, paraffin embedding, tissue sectioning, or staining.


To compare the ability of pathologists to evaluate direct-to-digital images with standard pathology preparations.


One hundred surgical pathology samples were obtained. Samples were first digitally imaged, then processed for standard histologic examination on 4-μm hematoxylin-eosin–stained sections and digitally scanned. The resulting digital images from both digital and standard scan sets were viewed by each of 4 reading pathologists. The data set consisted of 100 reference diagnoses and 800 study pathologist reads. Each study read was compared to the reference diagnosis, and also compared to that reader's diagnosis across both modalities.


The overall agreement rate, across 800 reads, was 97.9%. This consisted of 400 digital reads at 97.0% versus reference and 400 standard reads versus reference at 98.8%. Minor discordances (defined as alternative diagnoses without clinical treatment or outcome implications) were 6.1% overall, 7.2% for digital, and 5.0% for standard.


Pathologists can provide accurate diagnoses from fluorescence-imitating brightfield imaging slide-free images. Concordance and discordance rates are similar to published rates for comparisons of whole slide imaging to standard light microscopy of glass slides for primary diagnosis. It may be possible, therefore, to develop a slide-free, nondestructive approach for primary pathology diagnosis.

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Author notes

This work was supported by NIH grant 1R01EB028635 (Levenson and Fereidouni) and in part by Histolix.

Borowsky, Levenson, and Fereidouni are cofounders of Histolix Inc; Gown is a consultant to Histolix Inc; and Morningstar was a paid full-time employee of Histolix Inc for part of the time of the study. Fleury has a financial interest in a capital group that has invested in Histolix. The other authors have no relevant financial interest in the products or companies described in this article.