Abstract P6-04-18: Rapid diagnosis of breast biopsies with open-top light-sheet microscopy
Authors: Pope Brandy E. Olin , Dintzis Suzanne , Parker Elizabeth U. , Alvarez Rebeca , Alvarsson Alexandra , Rahbar Habib , Reder Nicholas 
Published: March 2023

Abstract
Intro: Detection of breast cancer is achieved through a diagnostic work-up that involves specialized breast imaging, image-guided biopsy, and pathological assessment. Pathology results typically require ~ 3 business days before a diagnosis is rendered, creating avoidable anxiety in women presenting with a breast abnormality. We describe a solution to deliver a rapid preliminary diagnosis within 30 minutes of biopsy. This rapid preliminary diagnosis has the potential to reduce anxiety, streamline patient care workflows, and reduce healthcare costs. Methods: Fresh 14 gauge breast biopsies in normal saline were received directly from the breast imaging clinic and immediately stained using the nuclear marker SYBR Gold (Invitrogen) and pan-protein marker Atto 655 NHS Ester (Sigma) prepared in dimethyl sulfoxide, washed, and cleared for imaging at a refractive index of 1.46 using 2,2’-Thiodiethanol (Sigma). The full process requires approximately 14 minutes for staining and clearing. After staining, we placed the biopsy in a custom-built specimen holder and imaged a 100-micron cross section along the full length of the biopsy using our custom open-top light-sheet microscope. Images were subsequently converted computationally to a standard hematoxylin and eosin (H&E) color format using Fiji and Aivia (Leica) software and were ready for evaluation by a pathologist on the same day they were collected. Results: Using the protocol above, we demonstrated the ability to stain, clear, image, and visualize needle core biopsies within 30 minutes of receiving the tissue sample. Processing the data and converting to the H&E color palette required additional time, often requiring 60-90 minutes, surpassing the overall 30 minute turnaround time goal. The images contained identifiable stroma, epithelial cells, immune cells, and duct structures to a depth of 100 microns. Discussion: We describe a method to obtain a microscopic image for preliminary diagnosis within 30 minutes of receipt of tissue. The quality of the images produced by the method shows promise for preliminary diagnosis. Additional optimization is needed in sample preparation and data processing to meet the 30 minute turnaround time requirement. This optimization can be achieved by parallelization of the data processing on a cluster or cloud to reduce the time by an order of magnitude, which is currently under investigation by our team. The imaging and data processing will also be accelerated by multi-resolution imaging, which will decrease the time of imaging and dataset size for processing. A diagnostic study, comparing the preliminary light-sheet-based diagnosis to the final formalin fixed paraffin embedded (FFPE) pathology, is underway.
Citation Format: Brandy E. Olin Pope, Suzanne Dintzis, Elizabeth U. Parker, Rebeca Alvarez, Alexandra Alvarsson, Habib Rahbar, Nicholas Reder. Rapid diagnosis of breast biopsies with open-top light-sheet microscopy [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P6-04-18.