Samples were held at −20°C for 24 h to bring samples to an appropriate temperature for cryosectioning. After being embedded in frozen section compound (VWR International, Westchester, PA), 5-μm thick cryosections were collected from the intestinal samples, transverse to the length of the intestine, using a Leica CM 1950 cryomicrotome. The cryosections were mounted on positively charged glass slides (VWR International; 5 cryosections per slide) and stored at 4°C for no more than 48 h before immunofluorescence staining.
Frozen section compound
Manufactured by Avantor
Sourced in Germany
The Frozen section compound is a product designed for the preparation of frozen tissue sections for microscopic analysis. It provides a medium for embedding and supporting tissue samples during the freezing process, ensuring the preservation of the sample's structural integrity. This compound is a key tool for researchers and clinicians working with frozen tissue samples in various fields, such as histology and pathology.
Automatically generated - may contain errors
4 protocols using frozen section compound
1
Intestinal Sample Cryosectioning Protocol
2
Histological Analysis of GAG and Collagen
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GAG and collagen production were visualized histologically after 14 days of culture. Formulation P‐0 or S‐50 was fixed for 10 minutes with 4% formaldehyde (Fisher Scientific), embedded in frozen section compound (VWR), snap‐frozen in methylbutane chilled with liquid nitrogen, and sectioned to 20 μm sections. Since the polymers tend to absorb the histological dyes non‐specifically, slides were rinsed with sodium citrate‐EDTA buffer at room temperature to remove the PNIPAAM‐g‐CS and alginate MPs by dissolution. Then, GAG and collagen were stained using 1% wt/vol alcian blue or 0.1% wt/vol picrosirius red, respectively. Cell nuclei were counterstained with Weigert's hematoxylin. ECM deposition was compared on days 0 and 14.
3
Cryohistological Analysis of Duodenum
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Samples were stored at −20°C for 24 h prior to cryohistological analysis. Duodenum samples were embedded in a frozen section compound (VWR International, Westchester, PA) and 5 μm-thick cryosections were collected transverse to the length of intestine using a CM 1950 cryomicrotome (Leica, Nussloch, Germany). Cryosections were then mounted on positively charged slides (VWR International; 5 cryosections per slide) and stored at 4°C for no more than 24 h before immunofluorescence staining.
4
Immunolabeling of Muscle Cells and Tissues
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Immunofluorescence labeling of cultured satellite cells, single EDL myofibers, and transverse sections of TA muscle was performed as described previously (Crist et al., 2009) . Pre-fixation was required for immunolabeling with antibodies against GFP. TAs were fixed for 2 hr in 0.5% paraformaldehyde at 4 C and equilibrated overnight in 20% sucrose at 4 C. Tissues were mounted in Frozen Section Compound (VWR) and flash frozen in a liquid nitrogencooled isopentane bath. For immunoblotting, cell lysates were prepared as described previously (Crist et al., 2009) . Densitometry of immunoblots was performed with ImageJ. EdU and OPP were detected by Click-IT Detection kits (Life Technologies). Apoptosis was detected by ApopTag Red In Situ Apoptosis Detection Kit (Millipore).
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