Confocal Microscope Measurement
Confocal Microscope Measurement - Related Videos
05:13
Selective Single-Neuron Laser Ablation in a Zebrafish Larva Using a Confocal Microscope
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2025
Source: Morsch, M., et. al., Triggering Cell Stress and Death Using Conventional UV Laser Confocal Microscopy. J. Vis. Exp. (2017) In this video, we explore the technique of high-intensity UV laser ablation for precisely targeting and studying individual neurons in transgenic zebrafish. We demonstrate how to prepare, immobilize, and visualize neuronal responses, highlighting the cellular dynamics of injury and recovery.
09:45
Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes
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Cited by 21 •
2011
This protocol describes a method to image fluorescent T cells introduced into lymph node slices. The technique permits real-time analyses of T cell migration with traditional widefield fluorescence or confocal microscopes.
08:32
Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis
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Cited by 3 •
2022
Extracellular vesicles (EVs) contribute to cellular biology and intercellular communications. There is a need for practical assays to visualize and quantify EVs uptake by the cells. The current protocol proposes the EV uptake assay by utilizing three-dimensional fluorescence imaging via confocal microscopy, following EV isolation by a nano-filtration-based microfluidic device.
09:46
Simultaneous pH Measurement in Endocytic and Cytosolic Compartments in Living Cells using Confocal Microscopy
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Cited by 20 •
2014
Several methods are available for measuring intracellular pH but very few of these allow simultaneous measurement of cytoplasmic and organellar pH. Here, we describe in detail a rapid and accurate methodology to simultaneously measure cytoplasmic and vesicular pH by ratiometric imaging of living cells.
11:17
Confocal Microscope-Based Laser Ablation and Regeneration Assay in Zebrafish Interneuromast Cells
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Cited by 13 •
2020
Laser ablation is a widely applicable technology for studying regeneration in biological systems. The presented protocol describes use of a standard laser-scanning confocal microscope for laser ablation and subsequent time-lapse imaging of regenerating interneuromast cells in the zebrafish lateral line.
FAQs
The pinhole acts as a spatial filter in front of the detector. Light originating away from the focal region is rejected, while signal from the focused location can pass through. This selective detection reduces background from out-of-focus regions, improving image contrast and depth resolution for quantitative examination of structures and surfaces.
Scanning a focused laser or other point source samples the specimen at defined locations rather than collecting an undifferentiated image from the entire field. Repeating this process across the specimen produces optical sections at different positions. These sections can be assembled into image stacks that support three-dimensional analysis of geometry and surface features.
The measurement can use emitted light from a specimen or reflected light from a surface, depending on the available signal. In either case, the detected light passes through the spatial-filtering path before reaching the detector. This allows the same measurement principle to examine internal structures or engineered surfaces without relying on physical sectioning.
Rejecting out-of-focus light separates signals from different optical depths more effectively than an image that combines them. The resulting sections provide clearer information about the location and shape of features. For engineering analysis, that improved separation supports measurements of surface form, microfabricated geometry, and dimensional relationships within a specimen.
A typical workflow positions the specimen for optical examination, focuses a point source, and scans it across the area of interest. Emitted or reflected light travels through the pinhole to the detector. Measurements collected at successive optical positions are then organized into image stacks for three-dimensional analysis, height mapping, or cross-sectional evaluation.
Image stacks can be processed to represent the measured specimen as a height map or cross-sectional view. A height map describes surface variation, whereas a cross-section reveals dimensional relationships through the measured region. These outputs convert optical sections into information that can support evaluation of structures, surfaces, and microscale features.
Engineers apply the method to surface topography measurement, dimensional inspection, material characterization, and analysis of microfabricated features. Because image stacks provide height and cross-sectional information without physically sectioning the sample, the technique can also support quality control, failure analysis, and microscale design evaluation.