Endothelial Cell Staining and Imaging

Staining was performed essentially as described previously (Yokomizo et al., 2019 (link), 2012 (link)). Staining was performed using anti-PECAM-1/CD31 antibody (Clone MEC 13.3, BD Pharmingen, 1:500) to detect endothelial cells and anti-RFP antibody (Rockland, 1:1000) to detect tdTomato. For immunofluorescent detection, either Cy3 or Alexa-647 conjugated secondary antibodies were used. All confocal microscopy was carried out on an Olympus FV1200 confocal equipped with GaAsP PMT detectors (Olympus).

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Koui Y., Ideue T., Boylan M., Anderson M.J., Osato M., Suda T., Yokomizo T, & Mukouyama Y.S. (2022). Hepatic leukemia factor-expressing paraxial mesoderm cells contribute to the developing brain vasculature. Biology Open, 11(9), bio059510.

Publication 2022

Clone MEC 13.3 anti-RFP antibody FV1200 confocal GaAsP PMT detectors

Anti antibody Antibodies Clone Confocal microscopy Endothelial cells Immunofluorescent Pecam 1 Tdtomato

Corresponding Organization :

Other organizations : National Heart Lung and Blood Institute, National Institutes of Health, Kumamoto University, National Cancer Institute, National University of Singapore, National University Cancer Institute, Singapore, Tokyo Women's Medical University

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Variable analysis

independent variables

Anti-PECAM-1/CD31 antibody (Clone MEC 13.3, BD Pharmingen, 1:500)
Anti-RFP antibody (Rockland, 1:1000)

dependent variables

Endothelial cell detection
TdTomato detection

control variables

Staining protocol (as described previously in Yokomizo et al., 2019 and 2012)
Confocal microscopy (Olympus FV1200 confocal equipped with GaAsP PMT detectors)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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