Quantifying Microvessel Density in CA1

For immunohistochemistry, the sections were incubated with mouse anti-CD31 (1:50, Thermo Scientific, MA1-80069, Waltham, United States) diluted in PBS overnight at 4°C after blocking with bovine serum albumin. Then the samples were incubated with a biotinylated secondary antibody followed by avidin-biotin-peroxidase complex. Positive staining was visualized with diaminobenzidine. The CA1 region was magnified for 40 times and 200 times, captured by a digital camera connected to a microscope (BX512DP70, Olympus, Tokyo, Japan). And the number of open microvessels was analyzed with Image-Pro Plus 5.0 software (IPP, Media Cybernetic, Bethesda, MD, United States). Five fields of CA1 region were examined for each animal (Tian et al., 2013 (link)).

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Gu Y.Y., Huang P., Li Q., Liu Y.Y., Liu G., Wang Y.H., Yi M., Yan L., Wei X.H., Yang L., Hu B.H., Zhao X.R., Chang X., Sun K., Pan C.S., Cui Y.C., Chen Q.F., Wang C.S., Fan J.Y., Ma Z.Z, & Han J.Y. (2018). YangXue QingNao Wan and Silibinin Capsules, the Two Chinese Medicines, Attenuate Cognitive Impairment in Aged LDLR (+/-) Golden Syrian Hamsters Involving Protection of Blood Brain Barrier. Frontiers in Physiology, 9, 658.

Publication 2018

mouse anti-CD31 BX512DP70

Animal Antibody Avidin Biotin Bovine serum albumin Ca1 fields Camera Immunohistochemistry Microscope Microvessels Mouse Peroxidase

Corresponding Organization : Shanghai Innovative Research Center of Traditional Chinese Medicine

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Variable analysis

independent variables

Mouse anti-CD31 antibody (1:50 dilution in PBS)

dependent variables

Number of open microvessels in the CA1 region

control variables

Incubation time (overnight at 4°C)
Blocking with bovine serum albumin
Incubation with biotinylated secondary antibody
Incubation with avidin-biotin-peroxidase complex
Visualization with diaminobenzidine
Magnification (40x and 200x)
Image analysis software (Image-Pro Plus 5.0)

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