Zinc formalin

Manufactured by Merck Group

Sourced in Germany, United States

Zinc formalin is a laboratory reagent used in the fixation and preservation of biological samples. It is a solution of zinc sulfate and formaldehyde in water. The primary function of zinc formalin is to maintain the structural integrity of cells and tissues, allowing for better microscopic examination and analysis.

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Lab products found in correlation

Low gelling temperature agarose, by Merck Group (1 mentions) 4 well culture slide, by BD (1 mentions) Deltavision ultra microscope, by GE Healthcare (1 mentions) Paraffin wax, by Leica (1 mentions) Tri reagent solution, by Thermo Fisher Scientific (1 mentions) Sudan black b, by Merck Group (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Surgipath paraplast, by Leica (1 mentions) Ds fi2, by Nikon (1 mentions) Pvdf filter, by Cytiva (1 mentions) Nta software, by Malvern Panalytical (1 mentions) Nanosight ns300, by Malvern Panalytical (1 mentions)

8 protocols using zinc formalin

Uterus Tissue Isolation and Preservation

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Female mice were euthanized and uteri were isolated. Collected samples were either frozen in liquid nitrogen for biochemical analysis or fixed in 10% zinc formalin (Sigma-Aldrich, St. Louis, MO) or frozen in OCT compound (Tissue-Tek Sakura, Torrance, CA) for tissue analysis. Both paraffin-embedded and OCT-embedded tissues were sectioned at 7 μm.

Shen J., Yao L., Lin Y.G., DeMayo F.J., Lydon J.P., Dubeau L, & Lee A.S. (2016). Glucose-regulated protein 94 deficiency induces squamous cell metaplasia and suppresses PTEN-null driven endometrial epithelial tumor development. Oncotarget, 7(12), 14885-14897.

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Pluripotency and β-cell Differentiation Imaging

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For pluripotency test, cell lines were cultured in 4-Well Culture Slide (Falcon) until 70% confluence was reached, then fixed with 4% paraformaldehyde. Terminally differentiated β cell aggregates were embedded in 2–4% of low gelling temperature agarose (Sigma-Aldrich), fixed with 10% zinc formalin (Sigma-Aldrich), then included in paraffin. For intracellular staining cells were permeabilized (PermWash 0.4% Triton X-100 in PBS) and stained by using the antibodies listed in the key resources table. All images were acquired using Confocal UltraVIEW ERS microscope (PerkinElmer Life Sciences) and DeltaVision Ultra microscope (GE Healthcare Life Sciences), then analyzed by using Fiji/ImageJ v.1.52p.

Chimienti R., Baccega T., Torchio S., Manenti F., Pellegrini S., Cospito A., Amabile A., Lombardo M.T., Monti P., Sordi V., Lombardo A., Malnati M, & Piemonti L. (2022). Engineering of immune checkpoints B7-H3 and CD155 enhances immune compatibility of MHC-I−/− iPSCs for β cell replacement. Cell Reports, 40(13), 111423.

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Cardiac and IMA Perivascular Adipose Tissue Isolation

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‘Cardiac’ perivascular adipose tissue (C-PVAT), located adjacent to the aortic root and around the coronary arteries, and PVAT surrounding the IMA (IMA-PVAT) was collected perioperatively and immediately transferred to the laboratory in ice-cold sterile saline solution (Braun). IMA-PVAT was collected immediately after preparation of the arterial bypass graft and before establishing the extracorporeal circulation. C-PVAT was collected during extracorporeal circulation, immediately after removing the aortic cross-clamp, from the same position in all patients, regardless of the localization of atherosclerotic lesions within the coronary tree. PVAT specimens were briefly rinsed with sterile saline and visible connective tissue or blood vessels removed. PVAT samples were then either directly processed under sterile conditions for the preparation of conditioned medium (CM) or stored at −80 °C pending protein or RNA isolation. Specimens intended for RNA isolation were transferred to TRI Reagent® Solution (ThermoFisher Scientific; Dreieich, Germany) before storage at −80 °C. PVAT samples intended for histology/immunohistochemistry were fixed in 10% zinc formalin (Sigma-Aldrich; Darmstadt, Germany), embedded in paraffin wax (Leica; Wetzlar, Germany) and cut into 5-μm-thick sections.

Drosos I., Pavlaki M., Ortega Carrillo M.D., Kourkouli A., Buschmann K., Konstantinou F., Gogiraju R., Bochenek M.L., Chalikias G., Tortopidis C., Vahl C.F., Mikroulis D., Tziakas D., Münzel T., Konstantinides S, & Schäfer K. (2019). Increased Lymphangiogenesis and Lymphangiogenic Growth Factor Expression in Perivascular Adipose Tissue of Patients with Coronary Artery Disease. Journal of Clinical Medicine, 8(7), 1000.

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Histological Characterization of Vascular Tissues

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Vascular tissues (carotid artery, aorta) were harvested from anesthetized mice and either fixed in 4% zinc formalin (Sigma-Aldrich,St. Louis, MO, USA) and embedded in paraffin (Surgipath^® Paraplast; Leica Biosystems Inc., Buffalo Grove, IL, USA) or in 4% paraformaldehyde (Sigma-Aldrich) followed by 30% and 15% sucrose and embedded in Tissue-Tek^® O.C.T. compound (Sakura via Science Services GmbH, Munich, Germany). Sections were stained with hematoxyline and eosin (H&E) to visualize cell nuclei (blue-black), with Verhoeff’s Elastica stain (VES) to visualize elastic fibers (black) or with a combined Masson Trichrome (MTC)–VES stain to visualize muscle fibers (red) and extracellular matrix (blue). Senescent cells were visualized using a kit to detect SA-β-Gal activity (Abcam) or sudan black B (Sigma; 0.1% in 70% ethanol) to detect the accumulation of lipofuscins, according to [52 (link)].

Schütz E., Gogiraju R., Pavlaki M., Drosos I., Georgiadis G.S., Argyriou C., Rim Ben Hallou A., Konstantinou F., Mikroulis D., Schüler R., Bochenek M.L., Gachkar S., Buschmann K., Lankeit M., Karbach S.H., Münzel T., Tziakas D., Konstantinides S, & Schäfer K. (2019). Age-Dependent and -Independent Effects of Perivascular Adipose Tissue and Its Paracrine Activities during Neointima Formation. International Journal of Molecular Sciences, 21(1), 282.

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Histological Analysis of Tumor and Lung

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Tumor and lung tissues were harvested and immediately fixed in zinc formalin (Sigma-Aldrich) by intratracheal instillation of fixative. Sectioning and standard hematoxylin and eosin (H&E) staining was performed by the Pathology Core Lab at the Children’s Hospital of Philadelphia Research Institute. Images were acquired using a Nikon DS-Fi2 digital microscope camera.

Diamond M.S., Lin J.H, & Vonderheide R.H. (2021). Site-dependent immune escape due to impaired dendritic cell cross-priming. Cancer immunology research, 9(8), 877-890.

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Lung Tissue Fixation and Histology

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Animals were anesthetized and transcardially perfused with PBS followed by zinc formalin (Sigma-Aldrich). Lungs were fixed in zinc formalin. For routine histology, tissue sections (∼4 µm each) were stained with hematoxylin and eosin.

Zhuang Z., Lai X., Sun J., Chen Z., Zhang Z., Dai J., Liu D., Li Y., Li F., Wang Y., Zhu A., Wang J., Yang W., Huang J., Li X., Hu L., Wen L., Zhuo J., Zhang Y., Chen D., Li S., Huang S., Shi Y., Zheng K., Zhong N., Zhao J., Zhou D, & Zhao J. (2021). Mapping and role of T cell response in SARS-CoV-2–infected mice. The Journal of Experimental Medicine, 218(4), e20202187.

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Lung Tissue Fixation and Sectioning

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Lungs were perfused with 1×PBS, fixed at 4°C in zinc formalin (Sigma) for 24 hours and subsequently embedded in paraffin. The paraffin blocks were cut in 5 µm sections by step-sectioning at 200 µm intervals.

Vandal G., Geiling B, & Dankort D. (2014). Ras Effector Mutant Expression Suggest a Negative Regulator Inhibits Lung Tumor Formation. PLoS ONE, 9(1), e84745.

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Nanoparticle Tracking Analysis of Extracellular Vesicles

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Particle concentration and size distribution was measured using nanoparticle tracking analysis (NTA) with the Nanosight (NS300; Malvern, Worcestershire, UK). NTA uses Brownian motion to calculate particle size. Prior to running any samples, air, water, and then air again was used to flush the tubing leading to the chamber where the video was captured. Water used for cleaning was filtered through a 0.02μm syringe filter (Whatman PVDF filter). Samples were loaded into the chamber at a syringe pump speed of 1000μl/min until the fluid filled the chamber. Standard measurements of 60s each at a speed of 35μl/min were perfomed in triplicate for each sample. Prior to diluting the samples, EV isolates were fixed with zinc formalin (Sigma-Aldrich, St. Louis, MO) at a 1:1 ratio. Dilutions of each sample were adjusted so that more than 10 particles were seen per frame without any particles visually overlapping (dilution factor ~150). Between samples, the tubing was flushed with air, water, and then air again. Total particle concentration and size distribution were calculated by the NTA software (Malvern Panalytical, Malvern, UK).

Nakashima K., Takizawa T., Ochiai W., Yanagisawa M., Hisatsune T., Nakafuku M., Miyazono K., Kishimoto T., Kageyama R, & Taga T. (2001). BMP2-mediated alteration in the developmental pathway of fetal mouse brain cells from neurogenesis to astrocytogenesis. Proceedings of the National Academy of Sciences of the United States of America, 98(10), 5868-5873.

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