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

Abstract

Microscopic examination of biopsied and resected prostatic specimens is the mainstay in the diagnosis of prostate cancer. However, conventional analysis of hematoxylin and eosin (H&E)-stained tissue is time-consuming and offers limited two-dimensional (2D) information. In the current study, we devised a method—termed Prostate Rapid Optical examination for cancer STATus (proSTAT)—for rapid screening of prostate cancer using high-resolution 2D and three-dimensional (3D) confocal images obtained after hydrophilic tissue clearing of 100-µm-thick tissue slices. The results of the proSTAT method were compared with those of traditional H&E stains for the analysis of cores ( n=15) obtained from radical prostatectomy specimens ( n=5). Gland lumen formation, consistent with Gleason pattern 3, was evident following tracking of multiple optical imaging sections. In addition, 3D rendering allowed visualizing a tubular network of interconnecting branches. Rapid 3D fluorescent labeling of tumor protein p63 accurately distinguished prostate adenocarcinoma from normal tissue and benign lesions. Compared with conventional stains, the 3D spatial and molecular information extracted from proSTAT may significantly increase the amount of available data for pathological assessment of prostate specimens. Our approach is amenable to automation and—subject to independent validation—can find a wide spectrum of clinical and research applications.

Schwann cells acquire a repair phenotype after assembling into spheroids and show enhanced in vivo therapeutic potential for promoting peripheral nerve repair

“…The treated sciatic nerves were harvested at 7 days postoperation. Additional fluorescent dye was added to stain the plasma membrane and nucleus, and the tissues were optically cleared in preparation for high‐resolution 3D confocal imaging 35,36 …”

“…Additional fluorescent dye was added to stain the plasma membrane and nucleus, and the tissues were optically cleared in preparation for high-resolution 3D confocal imaging. 35,36 One day after treatment, more CM-DiI-labeled cells were detected in the group that received SC spheroids than in the group that received SC suspensions (Figure 7a), suggesting that the acute retention rate of injected cells could be improved when cells were delivered as 3D spheroids. Seven days after cell administration, only a limited number of CM-DiI-positive cells were detected in the retrieved sciatic nerves treated with SC suspensions, revealing significant cell loss (Figure 7a and Supplementary Video 1).…”

Optimizing immunofluorescence with high-dynamic-range imaging to enhance PD-L1 expression evaluation for 3D pathology assessment from NSCLC tumor tissue

“…Regarding interobserver agreement between pathologists for TPS (Table 3 ), we found that the agreement of TPS in IF images after HDR processing was ameliorated, but still had not as much as the agreement in IHC, possibly because IF images were less frequently used in routine training of pathologists. Except for additional training and more clinical cases, we can also apply pseudo-H&E and IHC processing to transform the color domain of fluorescent images, which will help pathologists become more familiar with fluorescent images to evaluate the biomarker expression patterns in a familiar format 45 .…”