Ex Vivo Blood-Brain Barrier Permeability Imaging Analysis

Ex Vivo Blood-Brain Barrier Permeability Imaging Analysis

Blood-brain barrier (BBB) is a complex cellular system between brain tissue and blood, which is composed of closely connected endothelial cells and astrocyte projectiles. It can control the transport of certain substances in the blood circulation to the central nervous tissue, ensuring the stability of the internal environment of the central nervous tissue. The imbalance of blood-brain barrier can lead to serious neurological diseases, including multiple sclerosis, Alzheimer's disease, and cerebral ischemia. Therefore, imaging analysis of blood-brain barrier permeability is crucial for the development of neuroactive drugs.

Blood vessel staining using laminin-α4 enhances contrast and detection of mild BBB leakage eventsFigure 1. Blood vessel staining using laminin-α4 enhances contrast and detection of mild BBB leakage events (Nesbit M, et al. 2021).

Ex Vivo Blood-Brain Barrier Permeability Imaging Analysis

In vitro measurement of brain extravasation by intravenous injection of exogenous dyes such as fluorescein and Evans blue is a common method to determine BBB permeability in animal models. This method requires the use of non-physiological perfusion solutions, artificial blood vessel flow and pressure, and prolonged anesthesia time. There are potential confounding factors and require more mice to meet the research goals.

The immunodetection of endogenous proteins such as immunoglobulin G (IgG) and albumin extravasation of the brain can be used as a preferred alternative marker for BBB permeability, but pre-perfusion is required to remove intravascular fluorescence, or cumbersome identification and differentiation Fluorescent and substantial fluorescent proteins in blood vessels. In order to overcome these limitations, CD BioSciences provides a fast and accurate method to visualize and quantitatively analyze the BBB permeability in mouse and rat brain tissues, while avoiding the confounding effects of non-physiological conditions (such as perfusion), and eliminating any human related deviations from the analysis.

Ex Vivo Blood-Brain Barrier Permeability Imaging Analysis Workflow

The following is the general workflow of ex vivo blood-brain barrier permeability imaging analysis:

Step 1

Use a cryostat to slice the fast-frozen brain hemispheres and fix them with paraformaldehyde.

Step 2

Double immunofluorescence labeling with antibodies against mouse or rat IgG and laminin-α4.

Step 3

Image the sample on a confocal microscope and transfer the image to image analysis software for use.

Step 4

Intelligent segmentation of blood vessels and quantification of extravascular IgG through training machines.


Confocal imaging data set
Quantitative analysis of IgG extravasation

Image analysis parameter settings
Other relevant data

Our Advantages

  • Quickly and accurately process and analyze large data sets
  • High sensitivity detection
  • Avoid confounding effects of non-physiological conditions
  • Eliminate errors caused by human factors
  • Experienced scientists provide experimental consultation

CD BioSciences has professional technicians and advanced imaging equipment. We can provide you with fast and intelligent ex vitro blood-brain barrier permeability imaging analysis service, which reduces the burden of image analysis/processing for researchers, and is highly reliable and repeatable. If you have any needs, please feel free to contact us.


  1. Nesbit M, Mamo J C, Majimbi M, et al. Automated Quantitative Analysis of ex vivo Blood-Brain Barrier Permeability Using Intellesis Machine-Learning[J]. Frontiers in neuroscience, 2021, 15.

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Please note: Our services can only be used for research purposes. Do not use in diagnostic or therapeutic procedures!

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