Immunohistochemical Analysis of Rat Hearts

Tissues were processed as previously described (5 (link), 6 (link), 53 (link)). Briefly, rat hearts were excised under anesthesia after collection of echocardiographic data and perfused with 4% paraformaldehyde. Tissues were cryopreserved using 30% sucrose and embedded in OCT (optimal cutting temperature) (Tissue-Tek). Sections were cut to 7 μm using a cryostat and immunostained for isolectin B4 (Invitrogen) and α-SMA (Sigma). Cells and tissue sections were counterstained with DAPI nuclear stain (Sigma).

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Saha P., Sharma S., Korutla L., Datla S.R., Shoja-Taheri F., Mishra R., Bigham G.E., Sarkar M., Morales D., Bittle G., Gunasekaran M., Ambastha C., Arfat M.Y., Li D., Habertheuer A., Hu R., Platt M.O., Yang P., Davis M.E., Vallabhajosyula P, & Kaushal S. (2019). Circulating exosomes derived from transplanted progenitor cells aid the functional recovery of ischemic myocardium. Science translational medicine, 11(493), eaau1168.

Publication 2019

isolectin B4 α-SMA DAPI nuclear stain

Anesthesia Cells Dapi Echocardiographic Hearts Isolectin Paraformaldehyde Stain Sucrose Tissue

Corresponding Organization : University of Pennsylvania

Other organizations : The Wallace H. Coulter Department of Biomedical Engineering

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Variable analysis

independent variables

Perfusion with 4% paraformaldehyde
Cryopreservation using 30% sucrose
Embedding in OCT (optimal cutting temperature)

dependent variables

Immunostaining for isolectin B4 (Invitrogen) and α-SMA (Sigma)
Nuclear staining with DAPI

control variables

Rat hearts
Anesthesia
Echocardiographic data collection
Cryostat sectioning at 7 μm

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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