RNA FISH for neuroscience
Visualisation and quantification of neuroscience-related single RNA transcripts, using Stellaris™ RNA fluorescence in situ hybridization (RNA FISH) or smFISH, offers the unique ability to observe altered disease or region-specific expression in fixed cells or tissues, including an entire intact brain or organism.
Custom Stellaris Probe sets can be designed against almost any RNA target using our free online designer. We also offer positive control ShipReady probe sets and professionally designed DesignReady Probe sets, which undergo rigorous bioinformatic screening. The Stellaris buffer set streamlines your experiments and improves the quality of your images by decreasing troublesome background fluorescence.
Advanced neuroscience applications:
The breadth of information which can be gathered using Stellaris RNA FISH encompasses individual RNA transcript counting, transcription bursting within a single cell, and visualising wide bands of expression within an entire intact brain or organism. As the image above demonstrates, Stellaris RNA FISH probe sets can be used to differentiate between cell types in the brain and is completely compatible with either cells or tissue.
Whole brain imaging can relay an incredible amount of information about RNA expression. Stellaris has been published utilizing the whole tissue clearing technology CLARITY which allowed for visualisation of RNA distribution in an entire 3D intact mouse brain. Stellaris RNA FISH is also compatible with super resolution microscopy and whole mount preparations.
Stellaris RNA FISH resources for neuroscience
Professionally-designed neuro-specific probe sets include:
|DOHH||Necab1 and 2||PMP22||STUB1|
|GAD1 and 2||Neurog1||PROX1||UCP2|
Tyrosine Hydroxylase (green)
Honey Bee Brain whole mount
Courtesy of: Matthew McNeill and Gene Robinson
Neuro-specific support materials:
Fluorescent microscopy can be challenging in the brain due to high autofluorescence, so we’ve developed a specific protocol for use of Stellaris RNA FISH probe sets in mouse brain and a tech article to walk you through some applications.
Bitesize Bio Article: Challenges of Autofluorescence in Neuroscience
Highlighted neuroscience publications citing Stellaris RNA FISH Probe sets.
Over 33% of publications citing Stellaris RNA FISH publish in Cell, Nature, or Science
Image of simultaneous RNA FISH and immunofluorescence assay. eGFP RNA molecules (red), protein marker GFP (green), DAPI (blue) in mouse E13 primary DRG neuron culture cells (submitted by Adi Minis).
Visit our Citation Center for the complete list.
Multiplexed Intact-Tissue Transcriptional Analysis at Cellular Resolution. Emily Sylwestrak et al. Cell. 2016.
Deficiency of the Survival of Motor Neuron Protein Impairs mRNA Localization and Local Translation in the Growth Cone of Motor Neurons . Claudia Fallini et al. The Journal of Neuroscience. 2016.
Nuclear speckles are detention centers for transcripts containing expanded CAG repeats. Martyna Urbanek et al. BBA Molecular Basis of Disease. 2016.
Feedback Control of Gene Expression Variability in the Caenorhabditis elegans Wnt Pathway. Ni Ji et al. Cell. 2013.
Asymmetric Segregation of the Double-Stranded RNA Binding Protein Staufen2 during Mammalian Neural Stem Cell Divisions Promotes Lineage Progression. Gretchen Kusek et al. Cell Stem Cell. 2012.
Stellarvision™ Imaging Platform
Whole tissue section to single RNA transcripts in one image
Target RNA expression in regions of the brain and also perform single transcript counting in a single image. Below is an entire section of mouse brain probed with Stellaris RNA FISH probes against Plp1 mRNA (green) and imaged using the Stellarvision imaging platform. The resulting image allows you to zoom in on single transcripts from the full brain section view.
Zoomed in single molecule resolution
Plp1 mRNA labeled with Stellaris RNA FISH probes (green),
DAPI (red) and imaged with Stellarvision
(For more information on the Stellarvision imaging platform email Optical Biosystems.)