Open-top Bessel beam two-photon light sheet microscopy for three-dimensional pathology

Nondestructive pathology based on three-dimensional (3D) optical microscopy holds promise as a complement to traditional destructive hematoxylin and eosin (H&E) stained slide-based pathology by providing cellular information in high throughput manner. However, conventional techniques provided superficial information only due to shallow imaging depths. Herein, we developed open-top two-photon light sheet microscopy (OT-TP-LSM) for intraoperative 3D pathology. An extended depth of field two-photon excitation light sheet was generated by scanning a nondiffractive Bessel beam, and selective planar imaging was conducted with cameras at 400 frames/s max during the lateral translation of tissue specimens. Intrinsic second harmonic generation was collected for additional extracellular matrix (ECM) visualization. OT-TP-LSM was tested in various human cancer specimens including skin, pancreas, and prostate. High imaging depths were achieved owing to long excitation wavelengths and long wavele..., , , # Open-top Bessel beam two-photon light sheet microscopy for three-dimensional pathology

[https://doi.org/10.5061/dryad.pk0p2ngwn](https://doi.org/10.5061/dryad.pk0p2ngwn)

This dataset contains images and code that support the study entitled \"Open-top Bessel beam two-photon light sheet microscopy for three-dimensional pathology\" published in eLife ([https://doi.org/10.7554/eLife.92614.2)](https://doi.org/10.7554/eLife.92614.2\))

## Description of the data and file structure

This dataset contains OT-TP-LSM and H&E image datasets from skin (cancer), pancreas (normal, premalignant, cancer), prostate (benign, cancer). In addition, the pancreatic OT-TP-LSM and H&E images used to train cycle-consistent generational adversarial networks (CycleGAN) for virtual H&E are included.

**Image dataset of two basal cell carcinoma (BCC) skin cancer used in Figure 2 are represented in this database:**

SkinCancer_.zip and SkinCancer_H_E.zip

**Image dataset of normal and intraductal papil...