Diagnosing 12 prostate needle cores within an hour of biopsy via open-top light-sheet microscopy

Abstract

. Significance: Processing and diagnosing a set of 12 prostate biopsies using conventional histology methods typically take at least one day. A rapid and accurate process performed while the patient is still on-site could significantly improve the patient’s quality of life. Aim: We develop and assess the feasibility of a one-hour-to-diagnosis (1Hr2Dx) method for processing and providing a preliminary diagnosis of a set of 12 prostate biopsies. Approach: We developed a fluorescence staining, optical clearing, and 3D open-top light-sheet microscopy workflow to enable 12 prostate needle core biopsies to be processed and diagnosed within an hour of receipt. We analyzed 44 biopsies by the 1Hr2Dx method, which does not consume tissue. The biopsies were then processed for routine, slide-based 2D histology. Three pathologists independently evaluated the 3D 1Hr2Dx and 2D slide-based datasets in a blinded, randomized fashion. Turnaround times were recorded, and the accuracy of our method was compared with gold-standard slide-based histology. Results: The average turnaround time for tissue processing, imaging, and diagnosis was 44.5 min. The sensitivity and specificity of 1Hr2Dx in diagnosing cancer were both . Conclusions: The 1Hr2Dx method has the potential to improve patient care by providing an accurate preliminary diagnosis within an hour of biopsy.

Rapid Histological Assessment of Prostate Specimens in the Three-dimensional Space by Hydrophilic Tissue Clearing and Confocal Microscopy

“…By taking advantage of fluorescence counterstaining, we were able to acquire strong fluorescence signals with an emission spectrum distinct from that achievable with immunofluorescence. Although several fluorescence microscopy techniques based on the use of nucleic acid staining dyes have been described for assessing prostate needle biopsies, 23,24 the proSTAT workflow met the requirements for a complete analysis of the specimen with membrane staining and IHC. Specifically, the integration of a lipophilic tracer and p63 immunostaining was of great benefit not only for the distinction between malignant and benign lesions but also for avoiding alternative radiation-based techniques (e.g., microCT).…”

Slide Over

“…With additional multimagnification, multiimmersion, and rapid optical-clearing features, this optical design has demonstrated extraordinary performance and usability in 3D histopathology samples with a wide range of geometries. 64,78 Deep, Vibrant CellsdFull-Field Optical Coherence Tomography Fluorescence microscopy often depends on extrinsic staining for the generation of optical contrast. Although the staining procedures of most slide-free histopathology techniques are simple, fast, and minimally affect the quality of the sample, any extra manual steps present potential burdens in a clinical process.…”

Rapid and label-free histological imaging of unprocessed surgical tissues via dark-field reflectance ultraviolet microscopy

“… 6 , 7 , 8 Light-sheet microscopy (LSM) achieves optical sectioning by a thin “selective” illumination plane and the signal collection from the orthogonal direction. 9 , 10 , 11 , 12 Although these technologies can be applied to the pathological assessment of thick tissues and shorten turnaround time, fluorescence labeling is still required to provide image contrast. Fluorescence labeling is challenging to be integrated into the current clinical practice, especially intraoperative pathological examination.…”