Fluoromount g aqueous mounting medium

Manufactured by Southern Biotech

Sourced in United States

Fluoromount-G® is an aqueous mounting medium designed for the preparation of fluorescent microscopy samples. It is formulated to preserve the fluorescence of labeled samples and provide a long-lasting, clear, and durable mounting solution.

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Lab products found in correlation

Axio observer z1, by Zeiss (3 mentions) Superfrost plus microscope slide, by Thermo Fisher Scientific (1 mentions) Rhodamine labeled ricinus communis agglutinin 1, by Vector Laboratories (1 mentions) A11120, by Thermo Fisher Scientific (1 mentions) Streptavidin peroxidase conjugate, by Jackson ImmunoResearch (1 mentions) K3469, by Agilent Technologies (1 mentions) 90i microscope, by Nikon (1 mentions) Lsm 980 confocal microscope, by Zeiss (1 mentions) Cq1 confocal microscope, by Yokogawa (1 mentions)

Close spelling variants of this product

Fluoromount mounting medium Fluoromount-G medium Fluoromount-GTM Fluoromount-G FluoroMount medium Mounting medium Fluoromount

4 protocols using fluoromount g aqueous mounting medium

SARS-CoV-2 Infection in Hamster Organotypic Cultures

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Organotypic cultures from 7-day-old hamsters were infected with 1000 PFU of BetaCoV/France/IDF0571/2020 virus. In all, 24 h post infection, the slices were fixed during 1 h in 4% paraformaldehyde (PFA), washed in 1× Dulbecco’s phosphate-buffered saline (DPBS), and permeabilized and blocked in 1× DPBS-3% BSA- Triton X-100 (perm and block solution) overnight at 4 °C. Slices were incubated in the perm and block solution containing the primary antibodies overnight at 4 °C. After three washes (5 min each) in 1× DPBS, slices were incubated in the perm and block solution containing the secondary antibodies for 1 h at RT; donkey anti-rabbit conjugated with Alexa 488 or 555, donkey anti-mouse conjugated with Alexa 488 or 555, and donkey anti-goat conjugated with Alexa 555 or 647 antibodies (1:500 each) (Supplementary Table 3). After 3 washes in 1× DPBS, slices were mounted with Fluoromount-G® aqueous mounting medium (SouthernBiotech, catalog no. 0100-01) on epoxy slides (CEL-LINE, catalog no. 30-12A-BLACK-CE24) and coverslipped. Images were taken using an inverted microscope Zeiss Axio Observer. Z1 with confocal unit LSM 800 and analyzed using ImageJ software. All primary antibodies used in this study were validated previously for use in hamster tissue or the sequence homology of the epitopes was more than 85%.

Ferren M., Favède V., Decimo D., Iampietro M., Lieberman N.A., Weickert J.L., Pelissier R., Mazelier M., Terrier O., Moscona A., Porotto M., Greninger A.L., Messaddeq N., Horvat B, & Mathieu C. (2021). Hamster organotypic modeling of SARS-CoV-2 lung and brainstem infection. Nature Communications, 12, 5809.

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Choroidal Flatmount Staining Protocol

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Flatmounting and staining were performed according to a protocol described previously (Merrigan et al., 2020 (link); Sardar Pasha et al., 2018 (link)) and summarized here. After FA, 14 days after L-CNV induction, mice were euthanized by isoflurane overdose and eyes were harvested. The eyes were enucleated and fixed in 4% paraformaldehyde/PBS overnight. After dissection the posterior eye cups were prepared for choroidal flat mounts. The posterior eye cups were washed with PBS and permeabilized in blocking buffer containing 0.3% Triton X-100 and 5% bovine serum albumin (BSA) in PBS for 2 hours at 4°C. The eye cups were then double stained for vasculature with rhodamine-labeled Ricinus communis agglutinin I (Vector Laboratories, Burlingame, CA, USA) and Alexa Fluor 488-conjugated-Isolectin B4 from Griffonia simplicifolia (GS-IB4) (Molecular Probes/Thermo Fisher Scientific) at 1:250 dilution in buffer containing 0.3% Triton X-100, 0.5% BSA in PBS for 24–48 hours at 4°C. After antibody incubation, whole mounts were washed three times with PBS for 15 minutes. After washing, tissue was mounted in Fluoromount-G aqueous mounting medium (SouthernBiotech, Birmingham, AL, USA; Cat. No. 0100–01) to a Superfrost Plus Microscope Slide (Fisher Scientific) and cover-slipped.

Sishtla K., Lambert-Cheatham N., Lee B., Han D.H., Park J., Pasha S.P., Lee S., Kwon S., Muniyandi A., Park B., Odell N., Waller S., Park Y I.I., Lee S.J., Seo S.Y, & Corson T.W. (2022). Small molecule inhibitors of ferrochelatase are antiangiogenic agents. Cell chemical biology, 29(6), 1010-1023.e14.

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Immunohistochemical Analysis of Subretinal Injection

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Seventy-one days after subretinal injections, the animals were sacrificed. Eyes were enucleated and fixed in a solution of 4% paraformaldehyde, 20% isopropyl alcohol, and 4% zinc chloride for approximately 24 hours. Following fixation, each eye was cut vertically where the sub-retinal bleb had been, and the bisected eyes were fixed for another 24 hours in the same buffer. The tissue was then embedded in paraffin.
Paraffin sections were immunolabeled with primary rabbit anti-GFP antibody (1:500) (A-11120; Life Technologies) and a rabbit IgG control antibody (1:250, 1 µg/ml) (011-000-003; Jackson ImmunoResearch Laboratories, West Grove, PA). Slides were subsequently incubated with a secondary biotinylated donkey anti-Rabbit IgG antibody (1:400) (711-065-152; Jackson ImmunoResearch Laboratories), followed by a tertiary antibody, streptavidin-peroxidase conjugate (1:1,000) (016-030-084; Jackson ImmunoResearch Laboratories). Slides were then incubated with 3-amino-9-ethylcarbazole (AEC) and a high sensitivity substrate chromogen (K3469; DAKO, Carpinteria, CA), and counter stained with Mayer’s hematoxylin and mounted with Fluoromount-G aqueous mounting medium (0100-01; Southern Biotech, Birmingham, AL). Images were taken with a Nikon 90i microscope.

Choi V.W., Bigelow C.E., McGee T.L., Gujar A.N., Li H., Hanks S.M., Vrouvlianis J., Maker M., Leehy B., Zhang Y., Aranda J., Bounoutas G., Demirs J.T., Yang J., Ornberg R., Wang Y., Martin W., Stout K.R., Argentieri G., Grosenstein P., Diaz D., Turner O., Jaffee B.D., Police S.R, & Dryja T.P. (2015). AAV-mediated RLBP1 gene therapy improves the rate of dark adaptation in Rlbp1 knockout mice. Molecular Therapy. Methods & Clinical Development, 2, 15022-.

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Immunofluorescence Staining of Hamster OCCs

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OCCs from seven-day-old hamsters were fixed overnight in 4% methanol-free paraformaldehyde (PFA) and washed in 1× Dulbecco’s phosphate-buffered saline (DPBS). The slices were permeabilized and blocked in 1× DPBS-3% BSA-0.3% Triton X-100 (perm and block solution) for one hour at room temperature (RT). Slices were incubated in the perm and block solution containing the primary antibodies overnight at 4 °C. After 3 washes (for 5 mins each) in 1× DPBS, slices were incubated in the perm and block solution containing the secondary antibodies (1:750) and the DAPI (1:1000) for 2h at RT. After 3 washes in 1× DPBS, slices were mounted with Fluoromount-G^® aqueous mounting medium (SouthernBiotech, catalog no. 0100-01). Images were taken with the LSM980 confocal microscope (Zeiss, PLATIM Lyon, France) and the CQ1 confocal microscope (Yokogawa, PLATIM Lyon, France) and analyzed using ImageJ software v1.52. Primary and secondary antibodies and dilutions used are described in Table 1 below.

Ferren M., Lalande A., Iampietro M., Canus L., Decimo D., Gerlier D., Porotto M, & Mathieu C. (2023). Early Permissiveness of Central Nervous System Cells to Measles Virus Infection Is Determined by Hyperfusogenicity and Interferon Pressure. Viruses, 15(1), 229.

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