Systems and methods for serial staining and imaging
Inventors
Ragan, Timothy • Dimant, Chaim Moshe
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11519832-B2
Publication Date
2022-12-06
Expiration Date
Abstract
The present invention relates to systems and methods for sequential operation of staining, imaging and sectioning of tissue samples by a processing system. After each layer of the sample is removed by the sectioning system, the system automatically stains the exposed surface of a sample to a depth to enable imaging of the remaining tissue. The system then repeats the sectioning, staining and imaging steps in sequence to image the sample.
Core Innovation
The invention relates to serial blockface staining (SBS) for imaging a tissue sample by repeatedly performing sequential staining and sectioning cycles. An exposed region is stained after a tissue surface is sectioned, and the stain penetrates into the tissue sample to a penetration depth. Imaging is performed to generate images while the blockface is incrementally advanced through multiple sectioning and staining iterations.
A control system coordinates sequential staining, imaging and sectioning of the tissue sample, and controls sequential staining, imaging and sectioning so that the staining system adjusts the rate of penetration of the stain into the tissue sample. By reducing penetration time to reach a staining depth within the tissue sample, the workflow enables production of sequential images across the tissue volume. The control is connected to the imaging system, the sectioning system, and the staining system.
The described SBS workflow integrates sectioning and staining with hardware and staining components that support diffusion-enhancing or penetration-controlling behavior to shorten penetration time. The document describes diffusion-enhancing devices and configurations that include agitation, heating, sonication, electrophoresis arrangements, and histology microwave antigen retrieval, along with detergents and proteases. The approach is used with imaging modalities including multiphoton microscopy and other listed optical imaging techniques to produce three-dimensional datasets from serial sections.
Claims Coverage
The document provides two independent claims: a system claim and a method claim. Each independent claim centers on coordinating sectioning, staining, and imaging, while controlling stain penetration timing by adjusting penetration rate to reduce penetration time to a staining depth.
System for sequential staining, imaging and sectioning with controlled penetration rate
A system for imaging a tissue sample including an imaging system, a sectioning system that sections tissue slices from the tissue sample, a staining system operative to stain an exposed region with a stain in sequence with sectioning, wherein the stain penetrates to a penetration depth, and a control system connected to the imaging system, the sectioning system, and the staining system that controls sequential staining, imaging and sectioning and adjusts the rate of penetration to reduce penetration time to reach a staining depth.
Method for producing sequential images with iterated staining, imaging and sectioning under penetration-rate control
A method of producing sequential images of a tissue sample by staining a first surface with a stain to deliver the stain to a penetration depth, imaging to generate an image, removing a section to expose a second surface, and iterating the staining, imaging and removing steps in response to a control system wherein the control system adjusts the rate of penetration to control penetration time to a staining depth.
Across both independent claims, the inventive core is the coordinated sequential cycle of staining, imaging, and sectioning under a control system that adjusts stain penetration rate to reduce penetration time while achieving a targeted staining depth.
Stated Advantages
Reduce the penetration time of the stain to a staining depth within the tissue sample.
Documented Applications
Production of whole-organ 3D datasets using serial blockface staining with repeatedly exposed tissue surfaces and imaging across sequential sections.
Proof-of-concept imaging including NeuN immunostaining and DAPI small-molecule staining with serial sectioning spacing.
Interested in licensing this patent?