Why does null hypothesis testing matter for SEM-based choroid plexus validation?

Null hypothesis testing ensures that observed ultrastructural differences in choroid plexus morphology are statistically significant, supporting robust target validation and reducing false positives in early discovery.

How does independent variable isolation fit SEM tissue preparation workflows?

Isolating variables such as fixation time, dehydration gradients, and coating thickness allows teams to attribute morphological outcomes specifically to experimental manipulations, enhancing reproducibility and interpretability.

What do quantitative SEM measurements of microvilli enable in R&D?

Quantitative assessment of microvilli density and epithelial integrity provides objective benchmarks for comparing healthy and diseased tissue, informing model selection and translational alignment.

Why are replication requirements critical for cross-functional SEM studies?

Replication ensures that morphological findings are consistent across samples and operators, enabling reliable data sharing and decision-making between discovery, screening, and translational teams.

Which statistical analysis capabilities are required before SEM imaging implementation?

Teams must establish protocols for quantitative image analysis and statistical comparison of morphological features to ensure that SEM outputs support actionable R&D decisions and portfolio advancement.

Visualization of a Mouse Brain Choroid Plexus Using Scanning Electron Microscopy

JoVE Encyclopedia of Experiments

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0 просмотров • 2:23 мин • June 17th, 2025

Take the choroid plexus from a mouse brain in a specimen basket.

The tissue, located within the brain ventricles, contains a layer of epithelial cells featuring microvilli on their apical surface.

Incubate the tissue in a fixative solution to preserve its cellular architecture.

Wash with a buffer to remove excess fixative and stabilize the pH.

Incubate the tissue with osmium tetroxide that binds to cell membrane lipids, preserving membrane stability, then rinse off excess reagent.

Dehydrate the tissue using increasing alcohol concentrations to prevent tissue distortion during imaging.

Dry the tissue, secure it onto a carbon-coated specimen mount, then coat it with a conductive platinum layer.

Using scanning electron microscopy, focus an electron beam onto the surface. The platinum layer dissipates electrons, preventing charge buildup and minimizing imaging artifacts.

A detector captures the electrons backscattered from the surface, producing an image of the tissue exhibiting epithelial cells with microvilli.

To perform the sample preparation for SEM, transfer the freshly isolated choroid plexus into freshly made fixation solution, and incubate overnight at 4 degrees Celsius. Once done, wash the sample three times using 3 to 5 milliliters of 0.1 molar sodium cacodylate buffer for five minutes each.

Post-fix the samples in 3 to 5 milliliters of 2% osmium tetroxide in 0.1 molar sodium cacodylate buffer for 30 minutes. Then wash the samples three times for five minutes each using 3 to 5 milliliters of ultrapure water. Next, dehydrate the samples in an increasing series of ice-cold ethyl alcohol concentrations with 15 minutes per ethyl alcohol solution. Use a critical point dryer to dry the sample properly. Position the sample carefully on a specimen mount with a carbon sticker, and visualize the choroid plexus samples using SEM.