Fluoromount mounting medium

Manufactured by Novus Biologicals

Fluoromount mounting medium is a water-based, non-fluorescent, and optical-clearing solution used for mounting fluorescently labeled specimens on microscope slides. It is designed to preserve the fluorescent signal and maintain the structural integrity of the sample.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Streptavidin alexa fluor 488, by Thermo Fisher Scientific (1 mentions) D6694, by Merck Group (1 mentions) Tcs sp5 2 confocal microscope, by Leica (1 mentions) Tcs sp5 2, by Leica (1 mentions) Anti ambn, by Santa Cruz Biotechnology (1 mentions)

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Fluoromount

3 protocols using fluoromount mounting medium

Immunohistochemical Analysis of Dental Tissues

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Dental tissues were obtained for immunohistochemical analysis by ventricular perfusion of 4% PFA. Fixed hemimandibles were decalcified (10% EDTA, 2 weeks), embedded in paraffin, and sectioned (5 μm thick). Immunofluorescence was performed as reported [16 (link)] using the anti-DSCR1 antibody (D6694, SIGMA) using the biotin-labeled anti-rabbit IgG (1:500 dilution, Vector Laboratories), and detection was carried out using Streptavidin Alexa Fluor 488 (1:800 dilution, Life Technologies). Samples were embedded using Fluoromount mounting medium (Novus) containing DAPI (Thermo Fisher Scientific). Images were taken using a Leica TCS SP5 II confocal microscope.

Li Y., Costiniti V., Souza Bomfim G.H., Neginskaya M., Son G.Y., Rothermel B., Pavlov E, & Lacruz R.S. (2022). Overexpression of RCAN1, a Gene on Human Chromosome 21, Alters Cell Redox and Mitochondrial Function in Enamel Cells. Cells, 11(22), 3576.

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Immunofluorescence Analysis of Orai1 and Orai2 in Mandibles

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Orai1^K14 and Orai2^−/− mice were perfused with 4% paraformaldehyde, and mandibles were isolated, decalcified (10% EDTA) for 2 weeks, and embedded in paraffin to obtain thin sections (5 μm thick). Immunofluorescence staining was performed using an antigen retrieval step as described (69 (link)). The following primary antibodies (all rabbit-raised) were incubated at 4°C overnight: polyclonal rabbit anti-ORAI1 (1:75; S.F., New York University Medical Center) and monoclonal rabbit anti-ORAI2 [1:75; Abcam; clone EPR10043(2 (link))]. The next day, sections were washed twice for 5 min each with phosphate-buffered saline before incubation for 1 hour with biotin-labeled anti-rabbit immunoglobulin G (1:500 dilution; Vector Laboratories) and washing and incubation with streptavidin Alexa Fluor 488 (1:800 dilution; Life Technologies). Samples were embedded using Fluoromount mounting medium (Novus) containing DAPI (Thermo Fisher Scientific). Images were obtained using a Nikon Eclipse 2000TE or a Leica TCS SP5 II confocal microscope and edited using Icy BioImage analysis.

Eckstein M., Vaeth M., Aulestia F.J., Costiniti V., Kassam S.N., Bromage T.G., Pedersen P., Issekutz T., Idaghdour Y., Moursi A.M., Feske S, & Lacruz R.S. (2019). Differential regulation of Ca2+ influx by ORAI channels mediates enamel mineralization. Science signaling, 12(578), eaav4663.

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Immunofluorescence analysis of enamel proteins

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EO cells were isolated for immunohistochemical analysis and, after 24 h plating on 25 mm coverslips, fixed with 4% paraformaldehyde. Immunofluorescence staining was performed as follows: briefly, the cells were blocked and permeabilized with BSA 1%, Triton 0.1% in PBS for 5 min. After two washes in PBS, the following primary antibodies were used: anti-Ambn (1:500 dilution; SantaCruz sc-50534), and anti-AmelX (1:500 dilution; SantaCruz; sc-32892). After washing, detection was carried out using Alexa Fluor 488 (1:500 dilution; Life Technologies). Samples were embedded using Fluoromount mounting medium (Novus) containing DAPI. Images were taken using a Leica TCS SP5 II confocal microscope and edited using ImageJ (NIH).

Costiniti V., Bomfim G.H., Li Y., Mitaishvili E., Ye Z.W., Zhang J., Townsend D.M., Giacomello M, & Lacruz R.S. (2020). Mitochondrial Function in Enamel Development. Frontiers in Physiology, 11, 538.

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