Tissue tek embedding medium

Manufactured by Sakura Finetek

Sourced in United States

Tissue-Tek embedding medium is a histological embedding material used to prepare tissue samples for sectioning and microscopic analysis. It provides a firm, consistent support for the tissue during the sectioning process.

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Lab products found in correlation

Lsm800 confocal microscope, by Zeiss (2 mentions) Prolong gold with dapi, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Alexa fluor 488 green, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 red, by Thermo Fisher Scientific (1 mentions) 4 6 diamino 2 phenylindole dapi, by Thermo Fisher Scientific (1 mentions) Superfrost plus slide, by Thermo Fisher Scientific (1 mentions) Nile red, by Merck Group (1 mentions) Bx60 microscope, by Olympus (1 mentions) Sodium hydroxide (naoh), by Merck Group (1 mentions) Tetrahydrofuran, by Merck Group (1 mentions) Resomer rg 504 h, by Boehringer Ingelheim (1 mentions) Fisherbrand superfrostplus gold microscope slides, by Thermo Fisher Scientific (1 mentions) Rabbit igg isotype control, by Abcam (1 mentions) Cm1950, by Leica (1 mentions) Ha binding protein, by Merck Group (1 mentions)

Close spelling variants of this product

Tissue-Tek optimum cutting temperature embedding medium Tissue-Tek® optimal cutting temperature embedding medium Tissue-Tek OCT embedding medium Tissue-Tek medium Tissue-Tek Embedding medium

6 protocols using tissue tek embedding medium

Cryosectioning and Immunostaining of Murine Retina

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The superior cornea of the eyes was marked for orientation. To prepare samples for cryosectioning, the eyes were fixed in freshly prepared 4% paraformaldehyde solution overnight at 4 °C. After removal of the cornea and lens, the eyecup was washed in phosphate-buffered saline (PBS) three times, and transitioned through 5%, 10%, and 20% sucrose in PBS for 2 h each. Eyes were then embedded in an orientation-specific manner in the embedding mixture containing 1:1 ratio of Tissue-Tek embedding medium (Sakura Finetek, 4583) and 20% sucrose and then sectioned at 10-µm thickness using a cryostat. Retinal sections were blocked with 5% goat serum in PBS with 0.1% Triton-X100 (Sigma-Aldrich) and incubated with primary antibodies overnight at 4 °C. After washes, sections were incubated with secondary antibodies at room temperature for 1 h and then counterstained with ProLong Gold with DAPI (Invitrogen, P36941). Images were taken at comparable areas on sections with a fixed gain using Leica 6000 microscope (Leica Corp., Wetzlar, Germany) or a confocal microscope (Leica SP5, Leica Corp., Germany).

Qiu Y., Yao J., Jia L., Thompson D.A, & Zacks D.N. (2019). Shifting the balance of autophagy and proteasome activation reduces proteotoxic cell death: a novel therapeutic approach for restoring photoreceptor homeostasis. Cell Death & Disease, 10(8), 547.

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Immunofluorescent Labeling of Rat Mammary Tissue

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Mammary glands from rats on P5 were excised and fixed overnight in 4% paraformaldehyde, 5% sucrose, in PBS at 4 °C, followed by successive incubations at 4 °C in 30% sucrose in PBS (overnight), 30% sucrose in Tissue-Tek embedding medium (Sakura Finetek USA, Inc.) (30 min) and 100% Tissue-Tek (overnight). Cryosections (10–16 μm) were prepared and deposited on Superfrost Plus slides (Fisher Scientific, Pittsburgh, PA), post-fixed with 4% paraformaldehyde in PBS for 30 min, incubated in 0.2% Triton X-100 in PBS for 20 min, and blocked in 0.2% gelatin in PBS for 20 min. For labeling, both primary antibodies and secondary antibodies labeled with Alexa Fluor 488 (green) or Alexa Fluor 594 (red) (Molecular Probes, Eugene, OR) were incubated for 30 min at 37 °C, and coverslips were mounted onto slides with Moviol in the presence of 1 microgram/ml of 4,6-diamino-2-phenylindole (DAPI; Molecular Probes). Sections were visualized with an Axiovert 200 M deconvolution microscope (Carl Zeiss, Thornwood, NY). Images were collected and processed using SlideBook software (Intelligent Imaging Innovations, Inc., Denver CO). For double labeling, images in two channels were collected separately and then overlaid using Adobe Photoshop software (Adobe Systems Inc.). To label lipid, slides were incubated with 100 nM Nile red (Sigma) in PBS for 10 min.

Ladinsky M.S., Mardones G.A., Orlicky D.J., Howell K.E, & McManaman J.L. (2019). Electron Tomography Revels that Milk Lipids Originate from Endoplasmic Reticulum Domains with Novel Structural Features. Journal of mammary gland biology and neoplasia, 24(4), 293-304.

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Quantifying Tumor Hypoxia and Proliferation

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Tumors were removed and frozen in Tissue-Tek embedding medium (Sakura Finetek USA, Torrance CA) and sectioned to a 10 μm thickness on a cryotome. Sections were fixed for 10 minutes in 4% paraformaldehyde in PBS, rinsed three times in PBS, and blocked in 10% goat serum, 10% bovine serum albumin (BSA) in PBS for 1 hour. Sections were stained with FITC-conjugated anti-pimonidazole antibody (H&I Inc) diluted 1:20 in blocking solution for 1 hour. EdU was visualized with AlexaFluor 555 azide (Invitrogen), according to manufacturer's instructions. Immunofluorescent microscopic images were obtained using an Olympus BX60 microscope and Microsuite Biological Suite imaging software (Olympus America, Center Valley, PA). The fraction of the tumor staining positive for pimonidazole was calculated by applying Otsu thresholding in ImageJ. Necrotic tissue was readily identifiable on the sections based on its fragmented appearance in immunohistochemical images and was excluded from the analysis.

Hajj C., Russell J., Hart C.P., Goodman K.A., Lowery M.A., Haimovitz-Friedman A., Deasy J.O, & Humm J.L. (2017). A Combination of Radiation and the Hypoxia-Activated Prodrug Evofosfamide (TH-302) is Efficacious against a Human Orthotopic Pancreatic Tumor Model. Translational Oncology, 10(5), 760-765.

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PLGA Nanoparticle Encapsulation Protocol

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Equimolar uncapped poly(d,l-lactide-co-glycolide) (PLGA) was supplied by Boehringer Ingelheim (Ingelheim am Rhein, Germany) with the tradename (Resomer RG 504 H, Mw = 41.9 kDa, inherent viscosity of 0.5 dL/g). Bovine serum albumin, labeled with rhodamine as a fluorescent dye (BSA-Rhod, Mw = 66 kDa), was purchased from Molecular Probes Europe BV (Leiden, The Netherlands). Analytical grade dichloromethane (DCM) was provided by Carlo Erba (Milano, Italy). Tissue-Tek embedding medium was supplied by Sakura Finetek (Torrance, CA, USA). All other chemicals poly(vinyl alcohol, PVA, Mowiol 40–88; tetrahydrofuran, THF; polystyrene standards for GPC calibration; sodium hydroxide) from Sigma Aldrich (St. Louis, MO, USA) were used.

Netti P.A., Biondi M, & Frigione M. (2020). Experimental Studies and Modeling of the Degradation Process of Poly(Lactic-co-Glycolic Acid) Microspheres for Sustained Protein Release. Polymers, 12(9), 2042.

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Immunohistochemical Staining of Eyelid

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Eyelids were collected and immediately fixed in 4% buffered paraformaldehyde overnight and then embedded in Tissue-Tek embedding medium (Sakura Finetek USA, Inc., Torrance, CA, USA) for cryosectioning. Sections (10 μm) were cut using a Leica CM 1950 (Leica, Buffalo Grove, IL, USA) cryostat and collected on Fisherbrand SuperfrostPlus Gold microscope slides (Thermo Fisher Scientific). Upon use, sections were incubated for 30 minutes at 60°C, and excess tissue embedding medium was removed with PBS. Unspecific protein binding sites were blocked with 10% fetal bovine serum (FBS) prepared in PBS containing 0.01 M saponin. Sections were then incubated with the primary antibodies anti-Krt14 (PRB-155P; Covance, Princeton, NJ, USA) and HA binding protein (Millipore). Sections were washed and incubated with NeutrAvidin Alexa Fluor 555 conjugate and anti-rabbit produced in donkey conjugated with Alexa 488 for 1 hour at room temperature. A secondary control was carried out with rabbit IgG isotype control (Abcam, Cambridge MA, USA) in place of the primary antibody and did not yield any staining (results not shown). Slides were mounted in Fluoromount-G and imaged under an LSM 800 confocal microscope (Zeiss).

Sun M., Puri S., Parfitt G.J., Mutoji N, & Coulson-Thomas V.J. (2018). Hyaluronan Regulates Eyelid and Meibomian Gland Morphogenesis. Investigative Ophthalmology & Visual Science, 59(8), 3713-3727.

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Histological Evaluation of GAG Synthesis

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For histological evaluation of glycosaminoglycan (GAG) synthesis, cell pellets from each group were stained with Safranin-O and fast green staining at day 21. Staining was performed as described in our previous study [12 (link)]. Cell pellets were harvested, fixed in 4% paraformaldehyde for 4 h at room temperature, embedded in Tissue Tek embedding medium (Sakura Finetek, Torrance, CA, USA), and incubated for 20 min at −70 °C. Pellets were sectioned 5 mm thickness at −20 °C and mounted on slides. The sections were air-dried and post-fixed at −20 °C with acetone for 15 min.

Kim J., Bae H., Han H.S, & Lee J. (2023). Ultrasonic Enhancement of Chondrogenesis in Mesenchymal Stem Cells by Bolt-Clamped Langevin Transducers. Micromachines, 14(1), 202.

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