At the experimental endpoint, animals were sacrificed by an overdose of chloral hydrate (100 mg/kg, intraperitoneal injection) and were transcardially perfused with PBS, followed by 4% paraformaldehyde fixative solution. Once fixed, cervical spinal cord tissue (~C2–8) was harvested, post-fixed in 4% paraformaldehyde overnight, then cryoprotected in subsequent 12%, 18%, and 24% sucrose solutions. Tissue samples were then frozen over dry ice in Tissue-Tek® optimal cutting temperature embedding medium (Sakura Finetek USA Inc., Torrance, CA, USA) and stored at –80°C until further processing. Spinal cord cross-sections were cut using a cryostat at a 20-µm thickness.
Tissue tek optimal cutting temperature embedding medium
Manufactured by Sakura Finetek
Sourced in United States
Tissue-Tek® optimal cutting temperature embedding medium is a tissue freezing compound used for the cryosectioning of biological specimens. It is designed to provide optimal support and preservation of the tissue sample during the freezing and sectioning process.
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Lab products found in correlation
4 protocols using tissue tek optimal cutting temperature embedding medium
1
Spinal Cord Tissue Harvesting and Cryoprotection
2
Maritime Pine Seed Germination and Root Transcriptome Analysis
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Maritime pine seeds (P. pinaster Aiton) from "Sierra Segura y Alcaraz" (Albacete, Spain)
were provided by the Área de Recursos Genéticos Forestales of the Spanish Ministerio de Agricultura, Pesca y Alimentación. Seed germination was carried out following the protocol described elsewhere (Cañas et al., 2006
After three days of acclimation, the control group was irrigated with 80 mL of water and the experimental group was supplied with 80 mL of 3 mM NH 4 Cl. Root samples were collected 24 h after the treatment and immediately frozen in liquid N (Figure 1).
Ten seedlings were collected and pooled per each sample for RT-qPCR validation. The same experiment was carried out three independent times. The screening condition and adequate development of each experiment was verified through the gene expression analysis by RT-qPCR of control genes following previous results (Ortigosa et al., 2021) (Figure 1). To perform laser capture microdissection (LCM), seedling's root apexes were cut, and tissues (5-6 mm) were imbibed in a specimen holder with Tissue-Tek optimal cutting temperature embedding medium (Sakura Finetek, Alphen aan den Rijn, The Netherlands) and immediately frozen in liquid N for cryostat sectioning. Frozen samples were stored at -80°C until use.
were provided by the Área de Recursos Genéticos Forestales of the Spanish Ministerio de Agricultura, Pesca y Alimentación. Seed germination was carried out following the protocol described elsewhere (Cañas et al., 2006
After three days of acclimation, the control group was irrigated with 80 mL of water and the experimental group was supplied with 80 mL of 3 mM NH 4 Cl. Root samples were collected 24 h after the treatment and immediately frozen in liquid N (Figure 1).
Ten seedlings were collected and pooled per each sample for RT-qPCR validation. The same experiment was carried out three independent times. The screening condition and adequate development of each experiment was verified through the gene expression analysis by RT-qPCR of control genes following previous results (Ortigosa et al., 2021) (Figure 1). To perform laser capture microdissection (LCM), seedling's root apexes were cut, and tissues (5-6 mm) were imbibed in a specimen holder with Tissue-Tek optimal cutting temperature embedding medium (Sakura Finetek, Alphen aan den Rijn, The Netherlands) and immediately frozen in liquid N for cryostat sectioning. Frozen samples were stored at -80°C until use.
3
Cryosectioning of Mouse Eyecups for Retinal Layer Analysis
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Eye cups (n = 5) were embedded in Tissue-Tek Optimal Cutting Temperature (OCT) embedding medium (Sakura Finetek, Netherlands) after fixation with 4% paraformaldehyde in 0.1 M PB at 4°C for 90 minutes. Eyecups were sectioned along the vertical meridian on a cryostat at a thickness of 10 μm. TOPRO-3 (Invitrogen, Carlsbad, CA; T3605, dilution 1:1,000) was incubated for 10 minutes and washed for 30 minutes with 0.1M PB and cover-slipped with Vectashield mounting medium. The Zeiss LSM image browser software was used to measure the thickness of outer nuclear layer (ONL) and inner nuclear layer (INL). The measurements were taken within 1 mm from the optic nerve.
4
Amygdalar Tissue Preparation and Processing
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At seven days, animals were euthanized by transcardial perfusion of saline and 4% paraformaldehyde. Following collection, brains were stored in a 4% paraformaldehyde fixative solution. After 48 hours in fixative, the whole brains were placed in 30% sucrose solution for tissue sectioning preparation. Whole brains were embedded in Tissue-Tek® optimal cutting temperature (O.C.T.) embedding medium (Sakura Finetek USA, Inc., Torrance, CA) for cryostat processing in the coronal plane. Samples were then cut (40 µm) and sections containing amygdala nuclei were isolated (Bregma: −2.28 mm).
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