Immunohistochemical Analysis of Rat Hearts

Formaldehyde-fixed rat hearts were dehydrated and then embedded in paraffin. Paraffin-embedded sections (5 μm) were dewaxed and incubated with the primary antibody HIF-1α (Bioworld, catalog # BS3514), VEGF (Santa Cruz, catalog # sc-13083) or CD31 (Santa Cruz, catalog # sc-1506). After visualization with diaminobenzidine (DAB), images were acquired under the inverted Olympus IX70 microscope (Olympus Inc. Melville, NY). The CD31 immunohistochemical staining was performed to determine the capillary density. The myocardial capillary density (MCD) was also calculated as previously described 19 (link).

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Zhang L., Sun Z., Ren P., You M., Zhang J., Fang L., Wang J., Chen Y., Yan F., Zheng H, & Xie M. (2017). Localized Delivery of shRNA against PHD2 Protects the Heart from Acute Myocardial Infarction through Ultrasound-Targeted Cationic Microbubble Destruction. Theranostics, 7(1), 51-66.

Publication 2017

HIF-1α VEGF

Antibody Capillary Embedded paraffin Formaldehyde Hearts Microscope Myocardial Paraffin Vegf

Corresponding Organization :

Other organizations : Union Hospital, Huazhong University of Science and Technology, University Town of Shenzhen, Shenzhen University, Shenzhen Institutes of Advanced Technology

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Variable analysis

independent variables

Formaldehyde fixation of rat hearts
Dehydration of rat hearts
Embedding of rat hearts in paraffin
Dewaxing of paraffin-embedded sections
Incubation with primary antibodies: HIF-1α, VEGF, CD31

dependent variables

Visualization of HIF-1α, VEGF, and CD31 using diaminobenzidine (DAB)
Capillary density determined by CD31 immunohistochemical staining
Myocardial capillary density (MCD)

control variables

Thickness of paraffin-embedded sections (5 μm)
Microscope used to acquire images (inverted Olympus IX70 microscope)

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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