Tdtomato Imaging
Tdtomato Imaging - Related Videos
07:25
Optogenetic Activation of Zebrafish Somatosensory Neurons using ChEF-tdTomato
0 Views •
Cited by 6 •
2013
Optogenetic techniques have made it possible to study the contribution of specific neurons to behavior. We describe a method in larval zebrafish for activating single somatosensory neurons expressing a channelrhodopsin variant (ChEF) with a diode-pumped solid state (DPSS) laser and recording the elicited behaviors with a high-speed video camera.
11:58
In-vivo and Ex-vivo Calcium Imaging of an Olfactory Circuit Neuron in the Third-Instar Drosophila melanogaster Larva
0 Views •
2025
We present calcium imaging protocols for a Drosophila larval olfactory neuron, using a topical tissue adhesive for immobilization. This method enhances stability, facilitating reliable in-vivo and ex-vivo experiments. Custom R scripts analyze calcium signals, providing an efficient platform for detailed neurophysiological research.
13:45
Hybrid µCT-FMT imaging and image analysis
0 Views •
Cited by 34 •
2015
We describe a protocol for hybrid imaging, combining fluorescence-mediated tomography (FMT) with micro computed tomography (µCT). After fusion and reconstruction, we perform interactive organ segmentation to extract quantitative measurements of the fluorescence distribution.
04:02
Confocal Microscopy Imaging of Intact Dendritic Arbors and Spine Morphology in a Cleared Mouse Brain
0 Views •
2025
Source: Pekarek, B. T., et. al., Imaging and Quantification of Intact Neuronal Dendrites via CLARITY Tissue Clearing. J. Vis. Exp. (2021)In this video, we demonstrate the preparation, imaging, and analysis of a cleared mouse brain to examine dendritic arbors, spine morphology, and neuronal distribution using confocal microscopy and 3D reconstruction techniques.
09:08
Multiple-mouse Neuroanatomical Magnetic Resonance Imaging
0 Views •
Cited by 59 •
2011
Magnetic resonance imaging (MRI) has become an increasingly popular tool for examining the phenotype of genetically altered mice. This article illustrates the methods necessary to achieve high-throughput phenotyping of genetically altered mice using multiple-mouse MRI.
FAQs
Detectability depends on two linked events: the genetic construct must produce tdTomato, and its chromophore must mature before appropriate illumination can generate the red fluorescent signal. This means observed fluorescence reflects both expression and maturation, not simply the presence of the construct. Interpreting timing therefore requires distinguishing protein production from signal development.
The mature chromophore absorbs appropriate excitation light and then emits red fluorescence that a fluorescence microscope can detect. Signal location corresponds to where the tdTomato-bearing cell, protein, or structure is present in the sample. This optical relationship enables visualization of spatial organization while preserving the distinction between molecular position and genetic expression.
When attached to a protein through a genetic construct, tdTomato can act as a positional reporter for that protein. The resulting red signal helps researchers examine protein localization within cells or tissues rather than measuring only whether the construct was introduced. In biochemistry, this supports spatial analysis of protein behavior and its relationship to cellular organization.
A typical workflow starts with a genetic construct that enables tdTomato expression in the cells or structures of interest. After expression and chromophore maturation, the sample is illuminated with appropriate light, and fluorescence microscopy records the emitted red signal. The resulting image can then be interpreted according to the tagged target, reporter activity, or labeled cellular population.
Living-sample imaging uses the compatible red fluorescent signal to follow cell dynamics or biological processes over time. Imaging fixed samples instead emphasizes the location and organization of labeled cells, proteins, or structures at a preserved observation point. Selecting between these formats depends on whether the experiment prioritizes temporal behavior or spatial analysis of tissue and cellular organization.
The method can report gene activity, mark cell lineages, track cell dynamics, and reveal tissue organization. As a fusion tag, it contributes information about protein localization; as a cellular reporter, it indicates where genetically defined activity or populations occur. These uses allow biochemical studies to connect molecular signals with cellular and tissue-level structure.