Optical Sectioning Technique
Optical Sectioning Technique - Related Videos
06:22
Optical Sectioning and Visualization of the Intervertebral Disc from Embryonic Development to Degeneration
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2021
We present a method to investigate spatial chondrocyte organization in the anulus fibrosus of the intervertebral disc using an optical sectioning method.
03:14
Vibratome Sectioning of Whole Murine Kidney: A Technique to Generate Thin Sections of Murine Kidney
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2023
In this video, we demonstrate the vibratome sectioning of a whole mouse kidney to obtain thin slices. These sliced tissue sections can be stored at low temperatures until further downstream analysis.
08:57
Quantification of Optic Nerve Cross Sectional Area on MRI: A Novel Protocol using Fiji Software
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2021
We provided a detailed protocol for a standardized method of optic nerve assessment and quantification using MRI, utilizing a widely available imaging sequence, and open access software for image analysis. Following this standardized protocol would provide meaningful data for comparison between different patients and different studies.
07:34
Minimally-invasive Technique for Injection into Rat Optic Nerve
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Cited by 4 •
2015
Direct injection into the rat optic nerve is useful for regenerative research. We demonstrate a minimally-invasive technique for direct injection into a rat optic nerve that does not involve opening the skull. Using this method, surgical complications are minimized and recovery is more rapid.
07:55
The Optical Fractionator Technique to Estimate Cell Numbers in a Rat Model of Electroconvulsive Therapy
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Cited by 24 •
2017
Here, we present a stereological method, the optical fractionator, used to quantify the formation of new neurons, and their survival, in the rat hippocampus following electroconvulsive stimulation. When correctly implemented, the sensitivity and efficiency of stereological methods ensures accurate estimates with a fixed and predetermined precision.
FAQs
Pinhole-based systems use focused illumination and a pinhole to reject light originating outside the focal plane. Multiphoton systems achieve depth selectivity through excitation that is restricted to a localized region within the specimen. These mechanisms provide different routes to isolate information from a chosen depth, allowing investigators to examine internal cellular or tissue organization with improved image sharpness.
Light from structures above and below the focal plane can blur the signal assigned to the region being examined. Restricting image formation to the selected plane reduces this interference, so cellular arrangements, tissue organization, and lesion features appear more sharply separated. This focal discrimination is especially useful when complex biological specimens contain structures distributed through several depths.
The focal plane identifies the depth from which the system forms a sharp image. Changing that depth produces a sequence of distinct optical sections rather than relying on a single view of the specimen. When these sections are assembled, researchers can examine spatial relationships across the specimen and generate a three-dimensional representation of its internal organization.
Researchers acquire images at successive depths through the specimen, producing a series of optical sections that represents its three-dimensional structure. The sections can then be reconstructed into a volumetric view, preserving information about how cells, tissues, or lesion features are arranged relative to one another. This approach supports analysis of biological organization beyond what one image plane can show.
A typical workflow selects a specimen region, focuses imaging at a defined depth, and records a sharp section from that plane. The system then samples additional depths to create serial sections, which may be reconstructed for three-dimensional examination. Depending on the platform, depth selectivity comes from a pinhole-based arrangement or from multiphoton excitation.
This approach is useful when medical researchers need high-resolution views of cells, tissues, or disease-related changes while preserving information about depth. It supports microscopy studies, pathology research, and development of diagnostic approaches by revealing biological organization and lesion features. Its ability to image internal layers also contributes to investigations of minimally invasive imaging.
Optical sectioning can reveal how cellular and tissue structures are organized through different depths, rather than presenting only a surface or single focal view. In disease studies, serial sections may help visualize lesion features and their spatial relationships. These observations can inform pathology research, diagnostic development, and the interpretation of three-dimensional biological changes.