The tissue diagnosis of adenocarcinoma and intraductal carcinoma of the prostate (IDC-P) includes Gleason grading of tumor morphology on the hematoxylin and eosin (H&E) stain, and immunohistochemistry (IHC) markers on the PIN-4 stain (CK5/6, P63, AMACR). In this work, we create an automated system for producing both virtual H&E and PIN-4 IHC stains from unstained prostate tissue using a high-throughput multispectral fluorescence microscope and artificial intelligence & machine learning. We demonstrate that the virtual stainer models can produce high-quality images suitable for diagnosis by genitourinary pathologists. Specifically, we validate our system through extensive human review and computational analysis, using a previously-validated Gleason scoring model, and an expert panel, on a large dataset of test slides. This study extends our previous work on virtual staining from autofluorescence, demonstrates the clinical utility of this technology for prostate cancer, and exemplifies a rigorous standard of qualitative and quantitative evaluation for digital pathology.

Verily Life Sciences, LLC reports 3 pending patents on Virtual Staining and 1 on an AI-assisted Prostate Cancer Process. P.F.W., C.M., Y.W., J.P., C.S., M.G., A.H., K.N., P-H.C.C., T.J., E.W., D.F.S, P.C. are current or former employees of Alphabet Inc. (Verily and Google) and performed work for this study during their tenure at Alphabet. They also report equity ownership in Alphabet at the time of work. L.R., J.R. are current or former employees of Danaher Corporation (Leica Biosystems) and performed work for this study during their tenure at Danaher.

Verily Life Sciences, LLC reports 3 pending patents on Virtual Staining and 1 on an AI-assisted Prostate Cancer Process. P.F.W., C.M., Y.W., J.P., C.S., M.G., A.H., K.N., P-H.C.C., T.J., E.W., D.F.S, P.C. are current or former employees of Alphabet Inc. (Verily and Google) and performed work for this study during their tenure at Alphabet. They also report equity ownership in Alphabet at the time of work. L.R., J.R. are current or former employees of Danaher Corporation (Leica Biosystems) and performed work for this study during their tenure at Danaher.

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Yes

The details of the IRB/oversight body that provided approval or exemption for the research described are given below:

wcgIRB gave ethical approval for the study in this work.

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Yes

I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance).

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I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable.

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The data for the patient samples cannot be shared under the restrictions placed by the institutional review board. The data storage and retrieval infrastructure was built and managed by Google LLC.