Permafluor mounting medium

Manufactured by Thermo Fisher Scientific

Sourced in United States, China, United Kingdom

PermaFluor is a mounting medium designed for use in fluorescence microscopy. It is formulated to preserve the fluorescence of labeled samples and provide a durable, long-lasting seal for microscope slides.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (12 mentions) Lsm 700 confocal microscope, by Zeiss (6 mentions) Lsm 510, by Zeiss (4 mentions) Lsm 710 confocal microscope, by Zeiss (4 mentions) Axio scan z1 slide scanner, by Zeiss (3 mentions) Dm5500b microscope, by Leica (3 mentions) Rabbit anti dsred2, by Takara Bio (2 mentions) Vt1000, by Leica (2 mentions) Alexa fluor 568 goat anti rabbit, by Thermo Fisher Scientific (2 mentions) Masson s trichrome staining kit, by Abcam (2 mentions) Picro sirius red stain kit, by Abcam (2 mentions) Zen 2, by Zeiss (2 mentions) Vibratome vt1000, by Leica (2 mentions) Ta150041, by Merck Group (2 mentions) Lsm 700 confocal laser scanning microscope, by Zeiss (2 mentions) Avidin biotin blocking kit, by Vector Laboratories (2 mentions) Fv10i, by Olympus (2 mentions) Fv1000, by Olympus (2 mentions) Exosparkler exosome membrane labeling kit green, by Dojindo Laboratories (2 mentions) Plasmem bright red, by Dojindo Laboratories (2 mentions)

Close spelling variants of this product

PermaFluor (135 citations) Mounting medium (119 citations) PermaFluor aqueous mounting medium (68 citations) PermaFluor medium (7 citations) Vision™ PermaFluor™ Aqueous Mounting Medium (2 citations) Shandon PermaFluor mounting medium (2 citations) Permafluor permount mounting medium (2 citations)

47 protocols using permafluor mounting medium

Neural Crest Cell Fate Mapping

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All paraffin sections were stained with hematoxylin and eosin (H&E), using non-acidified hematoxylin. Samples were then imaged using an Olympus Vanox AHBT3 microscope.
For cell-fate mapping of neural crest cells, frozen sections were prepared for immunohistochemical analysis. All frozen sections were blocked with a 10% Donkey serum (Sigma-Aldrich Corp.) and 0.2% Triton X-100 (Sigma-Aldrich Corp.) in PBS solution for 30 min at room temperature. Control frozen sections were then incubated in a blocking buffer with a NC-marker with primary AP-2alpha monoclonal antibody 3b5–S (Developmental Studies Hybridoma Bank, Catalog # 3b5), diluted 1:100, overnight in 4°C and then incubated with secondary Alexa Fluor™ 594 goat anti-mouse IgG2b (y2b) (Thermo Fisher Scientific, Catalog # A-21145), diluted 1:250 with blocking solution for 1 hr. Then, control sections were counterstained with 4^′,6-diamidino-2-phenylindole (DAPI) for 10 min at room temperature, and then mounted with aqueous PermaFluor mounting medium (Thermo Scientific). Similarly, mutant sections were counterstained with 4^′,6-diamidino-2-phenylindole (DAPI) for 10 min at room temperature, and then mounted with aqueous PermaFluor mounting medium (Thermo Scientific). All fluorescent sections were then imaged using a Zeiss Observer D1 microscope.

Sanchez J., Miyake R., Cheng A., Liu T., Iseki S, & Kume T. (2020). Conditional inactivation of Foxc1 and Foxc2 in neural crest cells leads to cardiac abnormalities. Genesis (New York, N.Y. : 2000), 58(7), e23364.

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Immunohistochemical Analysis of Cell Polarization

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For in vitro immunohistochemical analysis, cells were plated on fibronectin (10 μg/ml)-coated glass-bottom dishes (Iwaki). For DCs, cells were stimulated with CCL19 for 30 min and fixed in 4% paraformaldehyde (PFA) for 20 min. For THP1, cells were activated with PMA (50 nM) for 2 days and then fixed in 4% PFA for 20 min. Cells were classified as polarized or non-polarized based on morphology. For histological analysis of the dermis of the ear, mouse ears were split into front and back, and then fixed in 4% paraformaldehyde for 20 min. After washes with PBS containing Triton X-100 (0.1% vol/vol), the samples were blocked with BSA (5% wt/vol) in PBS for 60 min, and then stained with primary antibodies at 4 °C overnight. After four washes with PBS containing 0.1% Triton X-100, the samples were stained with fluorescently labeled secondary antibodies for 1 h. The samples were mounted in PermaFluor mounting medium (Thermo Scientific). All fluorescence images were obtained by confocal microscopy (LSM 710, Zeiss). To evaluate co-localization, a Z-stack image was acquired by optimizing the pinhole size according to the objective lens (×63). Representative slice images are shown in the corresponding figures.

Nakatani T., Tsujimoto K., Park J., Jo T., Kimura T., Hayama Y., Konaka H., Morita T., Kato Y., Nishide M., Koyama S., Nada S., Okada M., Takamatsu H, & Kumanogoh A. (2021). The lysosomal Ragulator complex plays an essential role in leukocyte trafficking by activating myosin II. Nature Communications, 12, 3333.

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Immunohistochemical Visualization of Neuronal Markers

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Mice were anesthetized with Avertin and transcardially perfused with 4% paraformaldehyde (PFA). The brain was removed and fixed in 4% PFA/PBS for 2 hr at room temperature. Brains were then sectioned in the coronal plane into 100 μm thick slices using a vibratome (VT-1000, Leica). Slices were first blocked in 1% BSA with 0.3% triton X-100 in PBS for 1 hr at room temperature and then incubated with primary antibodies overnight at 4°C followed by incubation with secondary antibodies for 2 hr at room temperature. Slices were mounted in PermaFluor mounting medium (Thermo Scientific) and tiled z stack images were obtained using a laser scanning confocal microscope (Zeiss LSM510). The following primary antibodies were used: rabbit anti-dsRed2 (1:1000, Clontech) and chicken anti-GFP (1:1000, Abcam). The following secondary antibodies were used: Alexa Fluor 568 goat anti-rabbit (1:500 Thermo Fisher Scientific) and Alexa Fluor 488 goat anti-chicken (1:500 Thermo Fisher Scientific).

Tan H.L., Roth R.H., Graves A.R., Cudmore R.H, & Huganir R.L. (2020). Lamina-specific AMPA receptor dynamics following visual deprivation in vivo. eLife, 9, e52420.

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Immunofluorescence Microscopy of GLI2 and Centrosome

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Immunofluorescence microscopy was conducted as previously described (Dateyama et al., 2019 (link)). Briefly, cells were fixed with cold methanol for 5 min or 4% PFA in PBS for 10 min, and permeabilized with 0.2% Triton X-100/PBS for 10 min. Slides were blocked with 5% BSA/PBS prior to incubation with primary antibodies. Primary and secondary antibodies were diluted to the desire concentrations using 5% BSA/PBS. Secondary antibodies used were AlexaFluor350-, AlexaFluor488-, or AlexaFluor594- conjugated donkey anti-mouse, anti-rabbit, or anti-goat IgG (Invitrogen). Cells were stained with Hoechst33342 to visualize DNA. Mounted slides with PermaFluor Mounting Medium (Thermo Fisher Scientific) were observed and imaged using AxioObserver with a 63× lens. The pixel intensities of GLI2 in proximity to centrosome were quantified using ImageJ as previously described (Kobayashi et al., 2014 (link)).

Kobayashi T., Tanaka K., Mashima Y., Shoda A., Tokuda M, & Itoh H. (2020). CEP164 Deficiency Causes Hyperproliferation of Pancreatic Cancer Cells. Frontiers in Cell and Developmental Biology, 8, 587691.

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Immunohistochemistry of Mouse Motor Cortex

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Animals were anesthetized with Avertin (0.02ml/g) and then transcardially perfused with phosphate-buffered saline (PBS) and 4% paraformaldehyde (PFA). The brain was removed and post-fixed in 4% PFA/PBS for 2 hours. The brain was sectioned coronally into 100 μm thick slices using a vibratome (VT-1000, Leica). Free-floating sections underwent antigen retrieval using L.A.B. solution (Polysciences) and were blocked in 1% BSA with 0.3% triton X-100 in PBS for 1h at room temperature. Sections were incubated with primary antibodies overnight at 4°C and then with secondary antibodies either for 2 hours in room temperature. Washes after the primary and secondary antibody were done in PBS and antibodies were diluted in PBS containing 0.2% BSA. Slices from motor cortex were mounted in PermaFluor mounting medium (Thermo Scientific) and tiled z stack images were obtained using a laser scanning confocal microscope (Zeiss LSM510).
The following primary antibodies were used: rabbit anti-dsRed2 (1:1,000, Clontech) and rat anti-CTIP2 (1:250, Abcam). The following secondary antibodies were used: Alexa Fluor 568 goat anti-rabbit (1:500 Thermo Fisher Scientific) and Alexa Fluor 647 goat anti-rat (1:500 Thermo Fisher Scientific).

Roth R.H., Cudmore R.H., Tan H.L., Hong I., Zhang Y, & Huganir R.L. (2019). Cortical Synaptic AMPA Receptor Plasticity During Motor Learning. Neuron, 105(5), 895-908.e5.

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Perfusion-Based Immunohistochemistry in Mouse Brains

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Animals were deeply anesthetized to avoid any pain and sacrificed mechanically either by decapitation or exsanguination. All animal procedures were performed in accordance with the Virginia Tech guidelines for use and care of laboratory animals. Three-month-old mature adult mice were deeply anaesthetized using isoflurane, and mice hearts were cannulated and perfused with PBS (exsanguination) followed by 4 % paraformaldehyde (PFA). The perfused mice were decapitated and brains were dissected out and fixed in 4 % PFA overnight. Brains were cryopreserved by incubation in 30 % sucrose solution for 48 h. For sectioning, brains were embedded in CryoTek™ and 20 μm-thick cortical sections were generated using a cryostat (IEC). The cortical sections were immunostained as floating sections. Sections were permeabilized with 0.025 % Triton X-100 followed by blocking with 5 % goat serum. Sections were stained with primary antibodies for two hours followed by secondary antibody (1:500) incubation for 30 min. Finally, sections were mounted on slides using PermaFluor™ mounting medium (Thermo Scientific) and coverslips were sealed using nail polish.

Srivastava S., McMillan R., Willis J., Clark H., Chavan V., Liang C., Zhang H., Hulver M, & Mukherjee K. (2016). X-linked intellectual disability gene CASK regulates postnatal brain growth in a non-cell autonomous manner. Acta Neuropathologica Communications, 4, 30.

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Visualization of Tissue Autofluorescence

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Autofluorescence of tissues may be due to the accumulation of lipofuscin, an aging pigment that is stored in phagocytic cells (22 (link), 23 (link)). For visualization of autofluorescence in reproductive tissue section, slices were deparaffinized, rehydrated and embedded in aqueous PermaFluor™ mounting medium (#TA-030-FM, Thermo Fisher Scientific Inc., Waltham, MA, USA) with or without nuclear counterstaining in 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) solution (#D1306, Thermo Fisher Scientific).

Schröder S.K., Krizanac M., Kim P., Kessel J.C, & Weiskirchen R. (2024). Ovaries of estrogen receptor 1-deficient mice show iron overload and signs of aging. Frontiers in Endocrinology, 15, 1325386.

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Immunofluorescence analysis of conjunctival cells

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Filters containing the human conjunctival specimens were hydrated for 5 min with distilled water, permeabilized and blocked in a PBS solution containing 5% bovine serum albumin, 0.5% Tween-20 and 1% Triton X-100 for 1 h at room temperature. The cells were then incubated with anti-HSPA5 (1:100; #610978; BD Biosciences, Franklin Lakes, NJ, USA) or anti-DDIT3 (1:100; sc-7351; Santa Cruz Biotechnology) antibodies overnight at 4 °C. Samples were washed and incubated with the corresponding Alexa Fluor 555-conjugated secondary antibody (1:500; Thermo Fisher Scientific) and DAPI for 1 h at room temperature. Samples were mounted with a drop of Permafluor mounting medium (Thermo Fisher Scientific) and visualized using a Zeiss LSM 800 confocal microscope (Carl Zeiss MicroImaging GmbH, Jena, Germany). For cell culture experiments, cells were grown on glass chamber slides, washed with Dulbecco’s PBS, and fixed with 4% paraformaldehyde for 30 min. The slides were then coverslipped with antifade mounting medium containing DAPI (Vectashield; Vector Laboratories, Burlingame, CA, USA) and imaged using a Leica TCS SP5 confocal microscope (Leica Microsystems, Wetzlar, Germany).

Guindolet D., Woodward A.M., Gabison E.E, & Argüeso P. (2022). Alleviation of Endoplasmic Reticulum Stress Enhances Human Corneal Epithelial Cell Viability under Hyperosmotic Conditions. International Journal of Molecular Sciences, 23(9), 4528.

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Quantitative Histochemical Analysis of Liver Fibrosis

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FFPE liver sections (8 μm) were deparaffinized through xylenes and graded alcohols and then rehydrated in distilled water. The Masson’s trichrome staining kit (Abcam #ab150686) was used according to the manufacturer’s protocol. The Picro Sirius Red Stain Kit (Abcam #ab150681) was used to stain tissue sections for 60 min at room temperature. The slides were rinsed in two changes of acetic acid, three changes of ethanol, and then mounted using PermaFluor mounting medium (Thermo). Whole-section tiled images were acquired with an Axioscan.Z1 slide scanner (Zeiss) at 20× magnification, and quantification was performed with Zen 2.3 software (Zeiss).

Johnson N.R., Yuan P., Castillo E., Lopez T.P., Yue W., Bond A., Rivera B.M., Sullivan M.C., Hirouchi M., Giles K., Aoyagi A, & Condello C. (2023). CSF1R inhibitors induce a sex-specific resilient microglial phenotype and functional rescue in a tauopathy mouse model. Nature Communications, 14, 118.

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Immunohistochemistry of Fixed Brain Sections

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Fixed brains were sectioned to 30μm thickness using a Leica vibratome (VT1000S). For Ubiquitin, sections underwent heat-induced antigen retrieval at 95 °C for 30min (Buffer: 10mM sodium citrate, 0.05% Tween 20 (v/v), pH=6.0). For Aβ-antibody staining, sections were pre-treated with an antigen retrieval buffer containing 70% formic acid in 1x PBS for 10 minutes and washed with 1x PBS. Primary antibodies were diluted in saline with 0.2% (v/v) Triton-X 100 and 4% (v/v) normal goat serum (Jackson ImmunoResearch 005-000-121). Sections were incubated with the primary antibody solution at 4 °C for two days followed by secondary antibodies for 5h. For amyloid plaque staining, a Thioflavin S solution (0.0003% (m/m)) was used for 15min incubation at RT. Stained sections were mounted onto glass slides with PermaFluor mounting medium (Thermo Scientific, TA-030-FM).

Condello C., Yuan P., Schain A, & Grutzendler J. (2015). Microglia constitute a barrier that prevents neurotoxic protofibrillar Aβ42 hotspots around plaques. Nature communications, 6, 6176.

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