Skin and muscle incision were performed and ipsilateral and contralateral hindpaws were cut off at the ankle 1 or 4 days after surgery. The hindpaws were placed in Richard-Allan Scientific™ Buffered Zinc Formalin (Thermo Fisher Scientific) for at least 4 days. The hindpaws were removed from the formalin and cut into approximately equal thirds and placed in tissue cassettes in formalin overnight. The middle third of the hindpaw containing the incision was sectioned coronally and processed at the Children’s Hospital of Wisconsin Research Institute Histology Core Facility. Sections were stained with hematoxylin and eosin. A pathologist (M.W.L.) assessed the degree, character, and distribution of inflammation and tissue damage while blinded to genotype, treatment, and any a priori hypotheses. The Children’s Research Institute Imaging Core Facility used a Hamamatsu Nanozoomer HT slide scanner (Hamamatsu Photonics, K.K., Hamamatsu City, Japan) to generate images of tissue sections. The area of inflammation was manually calculated using NDP.View 2 software (Hamamatsu Photonics).
Richard allan scientific buffered zinc formalin
Manufactured by Thermo Fisher Scientific
Richard-Allan Scientific™ Buffered Zinc Formalin is a laboratory fixative solution used for the preservation and preparation of biological specimens for microscopic analysis. It contains a buffered mixture of zinc salts and formaldehyde. The solution aids in the preservation of cellular structure and morphology, enabling effective tissue fixation and preparation for subsequent histological processing and examination.
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3 protocols using richard allan scientific buffered zinc formalin
1
Histopathological Analysis of Hindpaw Inflammation
2
Perfusion and Fixation of Rodent Nervous Tissues
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The rats were ~ 4 months old when tissues were harvested. After transcardial perfusion with cold 100mL PBS, lumbar (L) 4 and 5 DRG, lumbar spinal cord, and sciatic nerve segments proximal to the sciatic bifurcation and terminal branches were dissected, and fixed in Richard-Allan Scientific™ Buffered Zinc Formalin (ThermoFisher) overnight, followed by processing for paraffin embedment. The previously described histological protocol was adopted.17 (link)
3
Tissue Collection and Processing for Histology and Western Blotting
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After transcardial perfusion with cold 100 mL PBS in rats and 40 mL in mouse, lumbar (L) 4 and 5 DRG lumbar spinal cord, and sciatic nerve segment proximal to the sciatic bifurcation were dissected, and fixed in Richard-Allan Scientific™ Buffered Zinc Formalin (ThermoFisher, Rockford, IL) overnight, followed by processing for paraffin embedment. The previously described histological protocol was adopted.26 (link)
For western blot experiments, L4/L5 DRG were removed, snap frozen in liquid nitrogen, and stored at −80°C for extraction of protein.
For western blot experiments, L4/L5 DRG were removed, snap frozen in liquid nitrogen, and stored at −80°C for extraction of protein.
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