Wga texas red x conjugate

For immunohistochemistry, hearts sections of 5–6μm thickness were co-stained with TUNEL and vWF using kit from Millipore (S7110) as previously described [5 (link),40 –43 (link)]. Secondary donkey anti-rabbit-647 alexa flour was used to detect vWF+ vascular structures. Fluorescence images (63×) were captured using a Zeiss LSM 880 confocal microscope (Carl Zeiss AG, Oberkochen, Germany) and analyzed using Zen 2011 software (Carl Zeiss AG). H&E staining and Masson’s trichrome staining was performed at the UIC histology core facility. Five fields from control and target heart sections were evaluated with an Olympus BX51/IX70 fluorescence microscope (Olympus, Tokyo, Japan). Morphometric analyses of the hearts were assessed using National Institute Health (NIH)-ImageJ software (Bethesda, MD, USA) as previously described [5 (link),40 –43 (link)]. To assess cardiomyocyte area, heart sections were stained with Wheat Germ Agglutinin (WGA)-Texas RedX conjugate (W21405, ThermoScientific, Waltham, MA). Paraffin section were de-parafinized, dehydrated, permeabilized with Triton-X 100 and incubated with 2μg/ml WGA-Texas Red-X conjugate, and washed. Images were examined and captured using BX51 Olympus microscope. Cardiomyocyte area was quantified using the NIH ImageJ software.