Protocol detail

Assessing Blood-Brain Barrier Integrity

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The sections made from rats selected using single-blinding were incubated with anti-occludin (Solarbio) and anti-ZO-1 (Solarbio) antibodies at 4°C overnight (Jin et al., 2021 (link)). After being washed with PBS, they were incubated together with the fluorescence-conjugated secondary antibody (Solarbio) at 25°Cfor for 1 h. In addition, the vascular endothelial cell markers CD31 were co-immunostaining with Zo-1 and occludin to observe the BBB integrity. The histopathological changes in the brain could be observed using a fluorescence microscope. The results were analyzed using ImageJ software.

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Xu S., Li X., Li Y., Li X., Lv E., Zhang X., Shi Y, & Wang Y. (2023). Neuroprotective effect of Dl-3-n-butylphthalide against ischemia–reperfusion injury is mediated by ferroptosis regulation via the SLC7A11/GSH/GPX4 pathway and the attenuation of blood–brain barrier disruption. Frontiers in Aging Neuroscience, 15, 1028178.

Publication 2023

Antibodies Brain Fluorescence antibody Fluorescence microscope Occludin Rats Vascular endothelial cell

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Variable analysis

independent variables

Incubation of brain sections with anti-occludin (Solarbio) and anti-ZO-1 (Solarbio) antibodies

dependent variables

Expression of occludin and ZO-1 proteins in the brain sections
Integrity of the blood-brain barrier (BBB)

control variables

Incubation time of brain sections with antibodies (4°C overnight)
Incubation time of brain sections with fluorescence-conjugated secondary antibody (25°C for 1 h)
Use of PBS for washing the brain sections

controls

Positive control: Co-immunostaining of vascular endothelial cell marker CD31 with ZO-1 and occludin to observe BBB integrity

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