Time Resolved Confocal Microscopy
Time Resolved Confocal Microscopy - Related Videos
02:07
Imaging Tumor Cell Migration in Glioblastoma Tissue Using Time-Lapse Scanning Confocal Microscopy
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2025
Source: Parker, J. J., et al. A Human Glioblastoma Organotypic Slice Culture Model for Study of Tumor Cell Migration and Patient-specific Effects of Anti-Invasive Drugs. J. Vis. Exp. (2017)This video showcases the imaging of glioblastoma tissue slices expressing green fluorescent protein (GFP) via confocal microscopy. High-resolution images are captured at consistent intervals to track tumor cell movement and direction.
10:07
Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers
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2014
High-resolution intravital imaging with enhanced contrast up to 120 µm depth in lymph nodes of adult mice is achieved by spatially modulating the excitation pattern of a multi-focal two-photon microscope. In 100 µm depth we measured resolutions of 487 nm (lateral) and 551 nm (axial), thus circumventing scattering and diffraction limits.
10:48
Imaging Biological Samples with Optical and Confocal Microscopy
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2023
Source: Peiman Shahbeigi-Roodposhti and Sina Shahbazmohamadi, Biomedical Engineering Department, University of Connecticut, Storrs, Connecticut Optical microscopes have been around for centuries, and while they reached their theoretical limitation of resolution decades ago, new equipment and techniques, such as confocal and digital image processing, have created new niches within the field of optical imaging. The best optical microscopes will typically have a resolution down to 200 nm in ideal...
10:05
Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy
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2021
Here, we provide a detailed protocol for the use of a rapid grid making device for both fast grid-making and for rapid mixing and freezing to conduct time-resolved experiments.
12:28
Correlative Super-resolution and Electron Microscopy to Resolve Protein Localization in Zebrafish Retina
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Cited by 11 •
2017
This protocol describes the necessary steps to obtain subcellular protein localization results on zebrafish retina by correlating super-resolution light microscopy and scanning electron microscopy images.
FAQs
Time-resolved detection converts photon arrival timing into a fluorescence-lifetime measurement. After pulsed excitation, the system records the distribution of delays before emitted photons reach the detector, rather than retaining only total brightness. The resulting decay behavior supplies a temporal signature for each fluorescent signal, allowing biological samples with similar intensity to be distinguished when their molecular environments differ.
The confocal pinhole and time-correlated detection contribute different kinds of selectivity. The pinhole suppresses fluorescence from planes outside the focal region, preserving spatial sectioning, while timing electronics sort detected photons by their arrival delay. Combining these measurements produces images that retain location and add lifetime information, so spatial context is not replaced by temporal analysis.
Fluorescence lifetime can reveal changes that intensity alone may miss. Two regions may emit comparable amounts of light yet produce different arrival-time patterns, indicating different local molecular environments or molecular states. This distinction is important because brightness provides a spatial signal, whereas lifetime adds a separate observable for interpreting biological organization and molecular state.
A typical acquisition links focused scanning with pulsed illumination and time-correlated photon detection. The laser scans the specimen through the confocal arrangement, emitted photons are collected, and their arrival delays are recorded for the scanned locations. Analysis then maps temporal behavior onto the image, generating lifetime-resolved information alongside the specimen’s spatial structure.
In biology, the measurements support fluorescence lifetime imaging and Förster resonance energy transfer analysis. They can be used to investigate protein interactions, ion signaling, and cellular dynamics because changes in fluorescence timing may report altered molecular surroundings or states. The method therefore connects optical measurements to questions about organization and activity within biological specimens.
Applied to ion signaling or cellular dynamics, the technique can follow fluorescence changes while preserving where they occur in the specimen. Its value lies in combining spatial localization with temporal readouts, enabling researchers to compare molecular behavior across cellular regions and distinguish dynamic or environmentally altered states that conventional intensity imaging may group together.