Tricaine buffer

Manufactured by Merck Group

Sourced in United States

Tricaine buffer is a laboratory product used to prepare solutions for the anesthesia of aquatic organisms, such as fish and amphibians. It serves as a buffering agent to maintain the pH of the solution within a specific range, which is essential for the effective and safe use of the anesthetic agent.

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Slicer, by Leica (1 mentions)

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Tricaine (556 citations)

4 protocols using tricaine buffer

Proteomic Analysis of Ricefield Eel Gonad Stages

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Wild ricefield eels (n = 210, body length = 39.42 ± 5.13 cm, and body weight = 48.53 ± 24.67 g) were purchased from a local market (Chengdu, Sichuan, China). The fish were maintained in the laboratory at a water temperature of 21.7 ± 2.5 °C under a photoperiod of 16 h light:8 h dark. Fish were decapitated after anesthesia with 0.02% tricaine buffer (80 µg/L) (Sigma, Saint Louis, MO, USA). Then, the gonads were collected and immediately stored in liquid nitrogen at − 80 °C. The gonads were divided into two parts. The first was fixed with Bouin’s solution for 24 h and then stored in 75% ethanol for determination of the gonadal developmental stage, while the second was immediately stored in liquid nitrogen at − 80 °C until RNA and protein extraction. Histological classification of the gonads, including OV, IE, IM, IL, and TE, has been described previously [16 (link)]. Three biological replicates were randomly selected from gonadal tissues at the OV, IE, IM, IL, and TE stages for proteome sequencing (Fig. 7).
Fig. 7

Identification of the gonad developmental stages in the ricefield eel Monopterus albus. A, ovary (OV). B, early intersex gonad (IE). C, middle intersexual gonad (IM). D, late intersex gonad (IL). E, testis (TE). CAO, cortical alveolar oocyte; PGO, primary growth stage oocyte; EVO, early vitellogenesis oocyte; GR, gonadal ridge; SC, spermatocyte; ST, spermatid

He Z., Xiao F., Yang D., Deng F., Ding W., He Z., Wang S., Chen Q., Wang H., Chen M., Gao K., Xiong J., Tang Z., Zhang M, & Yan T. (2024). Protein expression patterns and metal metabolites in a protogynous hermaphrodite fish, the ricefield eel (Monopterus albus). BMC Genomics, 25, 500.

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Reproductive Staging of Wild Mandarin Fish

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The wild M. albus (n = 100, body length = 34.93 ± 4.52 cm and body weight = 37.99 ± 21.88 g) used in the present study were obtained from a local market in Chengdu, Sichuan. These fish were kept under the natural temperature and photoperiod in the laboratory. All experimental procedures involving animal research were subject to approval and performed in accordance with the guidelines of the ethics committee (Approval No. 20190031).
Fish were anesthetized with 0.02% tricaine buffer (80 μg/L) (Sigma, West Hollywood, LA, USA) for 10 min after a 24 h fast, and the tissues, including half of the gonads, pituitary, eyes, heart, kidneys, intestines, spleen, muscles, and blood, were collected and immediately stored in liquid nitrogen. A portion of the fresh gonads was immediately fixed in Bouin’s solution for 24 h and embedded in paraffin. Sections were serially cut at a thickness of 5 μm using a slicer (Leica, Nussloch, Germany) and stained with hematoxylin/eosin. The histological classification of the gonad, including the PG, previtellogenic stage (PV), EV, MLV, and OM, has been described previously [44 (link)].

He Z., Chen Q., Xiong J., Chen M., Gao K., Lai B., Ding W., Huang J., Zheng L., Pu Y., Tang Z., Zhang M., Yang D, & Yan T. (2023). FoxH1 Represses the Promoter Activity of cyp19a1a in the Ricefield Eel (Monopterus albus). International Journal of Molecular Sciences, 24(18), 13712.

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Gonadal Development of Wild Ricefield Eels

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Wild ricefield eels (n = 210, body length = 29.42 ± 5.13 cm and body weight = 48.53 ± 24.67 g) were purchased from a local market (Chengdu, Sichuan, China). The fish were maintained in the laboratory at a water temperature of 21.7 ± 2.5 °C under a photoperiod of 16 h light:8 h dark. Fish were decapitated after anesthesia with 0.02% tricaine buffer (80 μg/L) (Sigma, Saint Louis, MO, USA) and the gonads were collected and immediately stored in liquid nitrogen at −80 °C. The gonads were divided into two parts: the first part was fixed with Bouin’s solution for 24 h and then stored in 75% ethanol for determination of the gonadal developmental stage, and the second part was immediately stored in liquid nitrogen at −80 °C until RNA and protein extraction.

He Z., Chen Q., He L., Xiong J., Gao K., Lai B., Zheng L., Pu Y., Jiao Y., Ma Z., Tang Z., Zhang M., Yang D, & Yan T. (2022). Cyt-C Mediated Mitochondrial Pathway Plays an Important Role in Oocyte Apoptosis in Ricefield Eel (Monopterus albus). International Journal of Molecular Sciences, 23(18), 10555.

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Gonadal Developmental Stages in Fish

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All adult fish were anesthetized with 0.02% tricaine buffer (80 µg/L) (Sigma, St. Louis, MO, USA) for 10 min after a 24 h fast. Tissues (n = 5), including the hypothalamus, telencephalon, epithalamus, heart, kidney, blood, liver, eyes, muscle, spleen, swim bladder, intestine, head kidney, medulla oblongata, skin, pituitary, gills, testes, and ovaries, were collected. Only the gonads were collected from the remaining individuals (n = 45). The tissues were snap-frozen in liquid nitrogen and stored at −80 °C until use. Fresh gonad samples were immobilized in Bouin’s solution and stored in 75% ethanol.
According to previous reports [25 (link),27 (link)], the gonadal developmental stages of fish were identified through histological analysis of the gonad sections (the 5 μm sections were stained with hematoxylin–eosin). The gonads were classified as follows: ovaries at the chromatin nucleolar stage (CNS), perinucleolar stage (PS), cortical alveoli stage (CAS), mid-vitellogenic stage (MVS), and late vitellogenic stage (LVS) and testes at the spermatogonium stage (SGT), early spermatogenic stage (EST), mid-spermatogenic stage (MST) and late spermatogenic stage (LST).

Yan T., Zhang S., Cai Y., Ma Z., He J., Zhang Q., Deng F., Ye L., Chen H., He L., Luo J., Yang D, & He Z. (2021). Estradiol Upregulates the Expression of the TGF-β Receptors ALK5 and BMPR2 during the Gonadal Development of Schizothorax prenanti. Animals : an Open Access Journal from MDPI, 11(5), 1365.

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