Ex Vivo Neuroimaging Techniques in Rodents

To analyze the entire mouse brain vasculature and investigate effects of ischemic stroke.

There was a missing ventral branch in the middle cerebral artery and remodeled anastomosis with dorsal arteries, showing reorientation of vessels towards the stroked site.

It is used for whole eye clearing to investigate the role of Dcc in optic nerve guidance during development.

<p>Anterograde axon tracing using AlexaFluor-555 and/or AlexaFluor-647-conjugated cholera toxin β subunit is used for visualization.</p> Signup and view all the answers

<p>The key regions include the optic chiasm, lateral geniculate nucleus, and superior colliculus.</p> Signup and view all the answers

<p>The primary antibody binds to the specific antigen in the sample.</p> Signup and view all the answers

<p>Colorimetric detection using HRP enzyme and DAB/H2O2 substrate.</p> Signup and view all the answers

<p>The secondary antibody binds to the primary antibody, amplifying the signal for detection.</p> Signup and view all the answers

<p>Blocking prevents non-specific binding of antibodies to the tissue.</p> Signup and view all the answers

<p>Alexa Fluor is a widely used fluorophore.</p> Signup and view all the answers

<p>A labeled secondary antibody provides a detectable signal to visualize the primary antibody-antigen complex.</p> Signup and view all the answers

<p>The protocol typically spans over 2 days.</p> Signup and view all the answers

<p>The enzyme catalyzes a reaction that produces a color change in the presence of its substrate.</p> Signup and view all the answers

<p>iDISCO+ utilizes optical clearing and 3D imaging techniques to visualize amyloid-β plaques.</p> Signup and view all the answers

<p>Amyloid-β plaques exhibit greater 3D complexity in humans compared to mice.</p> Signup and view all the answers

<p>Detergents and chemicals are used for permeabilization to facilitate antibody entrance into the tissue.</p> Signup and view all the answers

<p>Bleaching is employed to eliminate autofluorescence from the samples.</p> Signup and view all the answers

<p>Anti-beta amyloid antibodies (PAb #10) are used to label beta amyloid plaques.</p> Signup and view all the answers

<p>iDISCO+ facilitates the visualization of microglia enveloping the plaques using anti-Iba1 antibodies.</p> Signup and view all the answers

<p>Cell nuclei are labeled with TO-PRO-3 alongside beta amyloid plaques in the cortex.</p> Signup and view all the answers

<p>Surface rendering helps to isolate beta amyloid deposits along blood vessels.</p> Signup and view all the answers

<p>Ex vivo techniques involve analyzing tissues after the organism's death, while in vivo techniques observe brain function in living animals.</p> Signup and view all the answers

<p>Hematoxylin/eosin is a histochemical staining technique used to visualize brain morphology.</p> Signup and view all the answers

<p>Anesthetics and analgesia minimize pain and distress in the animals prior to euthanasia for brain study.</p> Signup and view all the answers

<p>A cryostat is used to cut freezing thin sections of the brain for analysis.</p> Signup and view all the answers

<p>3DISCO is used for whole brain clearing, allowing for visualization of brain structures in three dimensions.</p> Signup and view all the answers

<p>Immunohistochemistry allows researchers to visualize specific proteins or markers within tissue samples.</p> Signup and view all the answers

<p>Nissl staining helps in quantifying neurons and assessing brain morphology by highlighting cellular structures.</p> Signup and view all the answers

<p>Fixation preserves the brain's cellular architecture and prevents degradation during subsequent analysis.</p> Signup and view all the answers

<p>The primary purpose of uDISCO is to enable ultimate three-dimensional imaging of solvent cleared organs.</p> Signup and view all the answers

<p>The optimization takes advantage of tissue shrinkage to facilitate imaging of larger structures than before.</p> Signup and view all the answers

<p>Whole body clearing refers to the process of making an entire organism transparent for comprehensive imaging.</p> Signup and view all the answers

<p>Tracing individual axons allows for the determination of their entire trajectories within the central nervous system.</p> Signup and view all the answers

<p>The additional steps in iDISCO include permeabilization, bleaching, and blocking, followed by antibody incubations.</p> Signup and view all the answers

<p>Methanol is used to dehydrate the brain tissue during the clearing process in iDISCO.</p> Signup and view all the answers

<p>uDISCO uses only endogenous fluorescently labeled proteins for imaging purposes.</p> Signup and view all the answers

<p>The two primary components involved in the clearing process are methanol and dichloromethane.</p> Signup and view all the answers

<p>Doublecortin (DCX) is a marker for newly-formed neurons, indicating neurogenesis.</p> Signup and view all the answers

<p>The study found that stroke resulted in fewer new neurons, while the addition of Fortasyn increased the production of new neurons.</p> Signup and view all the answers

<p>In situ hybridization is used to detect specific mRNAs in tissue sections, allowing for the localization of gene expression in brain tissues.</p> Signup and view all the answers

<p>The steps include incubating with a DIG-labeled probe, incubating with an anti-DIG antibody labeled with alkaline phosphatase, and colorimetric detection using NBT/BCIP.</p> Signup and view all the answers

<p>The DIG-labeled probe binds to specific mRNAs in the tissue sections, enabling their detection during the hybridization process.</p> Signup and view all the answers

<p>An isocaloric control diet provides the same number of calories as the Fortasyn diet but lacks the specific nutrients that Fortasyn contains.</p> Signup and view all the answers

<p>The anti-DIG antibody binds to the DIG-labeled probe, facilitating subsequent detection and visualization of the target mRNA.</p> Signup and view all the answers

<p>Fortasyn enhances the production of newly-formed neurons compared to the control diet following a stroke.</p> Signup and view all the answers