Tbs tissue freezing medium

Manufactured by Thermo Fisher Scientific

Sourced in United States

TBS tissue freezing medium is a cryogenic medium designed for the preservation of biological samples, such as tissues and cells, during freezing and storage. It is formulated to protect the structural integrity and viability of the samples while undergoing the freezing process.

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Lab products found in correlation

Microm hm550, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Anti pcna, by Thermo Fisher Scientific (1 mentions) Mouse anti pcna, by Merck Group (1 mentions) Rabbit anti mef2, by Santa Cruz Biotechnology (1 mentions) Cm1850 cryostat, by Leica (1 mentions) Image pro express software, by Media Cybernetics (1 mentions) Dmi4000b, by Leica (1 mentions) Dmi4000b microscope, by Leica (1 mentions)

4 protocols using tbs tissue freezing medium

Cardiac Muscle Proliferation Quantification

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Hearts were extracted, fixed in 4% paraformaldehyde, equilibrated in 30% sucrose, then embedded in TBS tissue freezing medium (Fisher Scientific, Hampton, NH #TFM-C) and sectioned to 10 μm. Primary antibodies used were rabbit anti-Mef2 (Santa Cruz Biotechnology, Santa Cruz, CA #SC-313; 1:75) and mouse anti-Pcna (Sigma-Aldrich, Nadick, MA #P8825; 1:400). Secondary antibodies used were Alexa Fluor 488 goat anti-rabbit IgG (H + L) for anti-Mef2, and Alexa Fluor 594 goat anti-mouse IgG (H + L) for anti-Pcna (Invitrogen - Thermo Fisher Scientific, Waltham, MA #A11034, #A11020). Images were captured at 20× using an Olympus BX53 microscope and Retiga 2000DC camera. CM proliferation indices were calculated as a percentage of Mef2(+)Pcna(+) cells relative to the total number of Mef2(+) cells in a defined area adjacent to the injury.

FitzSimons M., Beauchemin M., Smith A.M., Stroh E.G., Kelpsch D.J., Lamb M.C., Tootle T.L, & Yin V.P. (2020). Cardiac injury modulates critical components of prostaglandin E2 signaling during zebrafish heart regeneration. Scientific Reports, 10, 3095.

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Cryosectioning of Embryos for Microscopy

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Embryos used for sectioning were incubated in successive 10%, 20%, and 30% sucrose in PBS solutions for a minimum of 3 hours each at room temperature, and then transferred to a 1:1 solution of TBS Tissue Freezing Medium (Fisher): 30% Sucrose in PBS for 8 hours. Embryos were then embedded in TBS Tissue Freezing Medium in disposable plastic moulds and frozen for 10 minutes using isopentane chilled with liquid nitrogen. Embryos were cryosectioned at 16 μm on a Leica CM1850 cryostat, dried for two hours at 35 °C on a slide warmer, and cover-slipped with 25% glycerol in PBS. Sections were imaged using a Zeiss AxioImager.M2 compound microscope equipped with an Axiocam 503 color camera and Zen 2012 (blue edition) software.

Tulenko F.J., Augustus G.J., Massey J.L., Sims S.E., Mazan S, & Davis M.C. (2016). HoxD expression in the fin-fold compartment of basal gnathostomes and implications for paired appendage evolution. Scientific Reports, 6, 22720.

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Histological analysis of muscle regeneration

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TA muscles were isolated and frozen in liquid nitrogen-cooled 2-methylbutane and embedded in TBS tissue freezing medium (Thermo Fisher Scientific, Waltham, MA, USA). Sections of 10 μm thickness were obtained with a Microm HM550 (Thermo Fisher Scientific, Waltham, MA, USA) at −20°C, placed on uncoated slides, and stained with hematoxylin and eosin (H&E). Separately, the sections from D3AI were fixed by 1.5% paraformaldehyde, incubated with anti-Ki-67 antibody, followed by incubation with DAPI and Alexa-conjugated secondary antibody. Images were obtained by DMI 4000B fluorescence microscope (Leica, Wetzlar, Germany) with a 20X dry objective (Fluotar, numerical aperture 0.4; Leica). Five bright field or fluorescence images were randomly captured using RETIGA EXi camera (QImaging, Surry, BC, Canada) and Image Pro Express software (Media Cybernetics, Rockville, MD, USA). The images were analyzed for cross-sectional area of all centrally nucleated regenerating myofibers (H&E staining) or counting Ki-67-positive nuclei using ImageJ. All procedures were performed by an investigator that were blinded to sample identification.

Kim D., Singh N., Waldemer-Streyer R.J., Yoon M.S, & Chen J. (2020). Muscle-derived TRAIL negatively regulates myogenic differentiation. Experimental cell research, 394(1), 112165.

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Histomorphometric Analysis of Regenerating Muscle

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Isolated TA muscles were frozen in liquid nitrogen-cooled 2-methylbutane and embedded in a TBS tissue freezing medium (Thermo Fisher Scientific). Sections of 10 μm thickness were made with a Microm HM550 (Thermo Fisher Scientific) at −20°C, placed on uncoated slides, and stained with hematoxylin and eosin (H&E). Five to ten images of the injured areas were randomly captured from the stained sections with a 20× dry objective (Fluotar, numerical aperture 0.4; Leica) on a Leica DMI 4000B microscope. The images were then analyzed for cross-sectional area (CSA) of centrally nucleated regenerating myofibers. A total area of ~750 000 μm² from the regenerating regions of each TA muscle was subjected to the measurement using ImageJ (NIH). Because injection was always performed with shScramble in one leg and shThrRS in the other leg, the data were paired, with shScramble as reference.

Kasahara S., Wang P, & Nuss D.L. (2000). Identification of bdm-1, a gene involved in G protein β-subunit function and α-subunit accumulation. Proceedings of the National Academy of Sciences of the United States of America, 97(1), 412-417.

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