Prolong glass mounting media

Manufactured by Thermo Fisher Scientific

Prolong Glass mounting media is a water-based, glycerol-based mounting medium designed for the preservation of fluorescent signals in microscopy samples. It is optimized to reduce photobleaching and maintain the clarity and intensity of fluorescent labels.

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Lab products found in correlation

Lsm800 confocal microscope, by Zeiss (4 mentions) Dapi solution, by Thermo Fisher Scientific (2 mentions) Normal rat serum, by Thermo Fisher Scientific (1 mentions) Prolong diamond, by Thermo Fisher Scientific (1 mentions) Tcs sp8 sted microscope, by Leica (1 mentions) Las x, by Leica (1 mentions) Imaris, by Oxford Instruments (1 mentions) Observer z1 inverted microscope, by Zeiss (1 mentions)

Close spelling variants of this product

ProLong glass antifade mounting media (5 citations) ProLong mounting media (9 citations) Prolong Glass (58 citations) Mounting media (34 citations)

4 protocols using prolong glass mounting media

Immunofluorescence Imaging of Immune Cells in Mouse Omentum

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Whole omenta were harvested from mice and fixed in 1% PFA overnight at 4°C. After rinsing, tissue was blocked using 10% BSA, 0.5% normal rat serum (Invitrogen), and 1 μg/ml 2.4G2 (BD) in PBS for 1 hr at room temperature. For immunofluorescence staining, omenta were incubated in PBS containing primary antibodies at 4°C for 3 days and subsequently rinsed with PBS overnight. Antibodies included: B220: AF647, BD Biosciences: 557683, clone: RA3-6B2; B220: BV510, BioLegend: 103248, clone: RA3-6B2; CD11b: AF700, eBioscience: 56-0112-82, clone: M1/70; CD3: AF532, Invitrogen: 58-0032-82, clone: 17A2; CD4: AF532, Invitrogen: 58-0042-82, clone: RM4-5; CD8b: ef450, Invitrogen: 48-0082-82, clone: eBioH35-17.2; ER-TR7: AF647, Santa Cruz Biotechnology: sc-73355 AF647, clone: ER-TR7. Tissues were then mounted on slides using Prolong Diamond (Invitrogen: P36970) or Prolong Glass mounting media (Invitrogen: P36984) and imaged using a Leica TCS SP8 STED microscope and acquired using LAS X (Leica Microsystems). Images were processed and analyzed using LAS X and Imaris (Oxford Instruments).

Christian D.A., Adams TA I.I., Shallberg L.A., Phan A.T., Smith T.E., Abraha M., Perry J., Ruthel G., Clark J.T., Pritchard G.H., Aronson L.R., Gossa S., McGavern D.B., Kedl R.M, & Hunter C.A. (2022). cDC1 coordinate innate and adaptive responses in the omentum required for T cell priming and memory. Science immunology, 7(75), eabq7432.

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Visualizing mRNA Decapping Complexes

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Transfected cells were washed once with PBS then fixed in 4% paraformaldehyde for 30 min. Cells were blocked and permeabilized in 10% normal goat serum and 0.1% Triton-X100 in PBS for 1 h. Cells were then stained with an antibody against enhancer of mRNA decapping 4 (EDC4) (1:200, Abcam) at room temperature for 1 h before washing three times with PBS. Secondary antibody (Invitrogen) was then applied for 1 h, slides washed three times with PBS, and incubated with DAPI solution (Invitrogen) to stain nuclei. Slides were coverslipped with Prolong Glass mounting media (Invitrogen) and stored at −20°C until imaged using a Zeiss LSM800 confocal microscope. ImageJ was used to automate EDC4⁺ puncta counts in GFP+ cells to remove bias.

Shaw B.C, & Williams J.L. (2024). A novel PSMB8 isoform associated with multiple sclerosis lesions induces P-body formation. bioRxiv.

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Quantifying EDC4 Puncta in Transfected Cells

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Transfected cells were washed once with PBS then fixed in 4% paraformaldehyde for 30 min. Cells were blocked and permeabilized in 10% normal goat serum and 0.1% Triton-X100 in PBS for 1 h. Cells were then stained with a knockout-validated antibody against enhancer of mRNA decapping 4 (EDC4) (1:200, Abcam) (Hallacli et al., 2022 (link)) at room temperature for 1 h before washing three times with PBS. Secondary antibody (Invitrogen) was then applied for 1 h, slides washed three times with PBS, and incubated with DAPI solution (Invitrogen) to stain nuclei. Slides were coverslipped with Prolong Glass mounting media (Invitrogen) and stored at -20°C until imaged using a Zeiss LSM800 confocal microscope. ImageJ was used to automate EDC4⁺ puncta counts in GFP⁺ cells to remove bias.

Shaw B.C, & Williams J.L. (2024). A novel PSMB8 isoform associated with multiple sclerosis lesions induces P-body formation. Frontiers in Cellular Neuroscience, 18, 1379261.

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Imaging GSDMB localization in Shigella-infected cells

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H1299 cells (WT or stably expressing GSDMB-FLAG) were plated at 50,000 cells/well on glass coverslips in a 24-well plate. Cells were infected with Shigella (MOI = 100) using a standard infection protocol (previously described). Cells were incubated with bacteria for 1.5 hrs, washed once with complete media, incubated with media containing 50 μg/ml gentamicin for 1 hr, washed again and incubated an additional 2hrs in media containing 50 μg/ml gentamicin. Cells were then washed twice with PBS and fixed with 4% PFA (in PBS) for 10 min. at room temperature. After fixation, cells were washed 3 times with PBS, permeabilized in 0.5% TX-100 + 1% BSA (in PBS) for 5 min., washed twice with PBS and blocked in 1% BSA (in PBS) for 30 min. Cells were incubated with rabbit anti-GSDMB primary antibody (1:1000, abcam 215729) 1 h at RT, washed 3 times in PBS and then incubated with goat anti-rabbit TX Red (1:200, Invitrogen T2767) for 1 h. After antibody incubation, cells were washed 3x with PBS, once with water and coverslips were mounted on slides with Prolong Glass mounting media (Invitrogen P36980) and imaged on an Observer.Z1 inverted microscope (Zeiss).

Hansen J.M., de Jong M.F., Wu Q., Zhang L.S., Heisler D.B., Alto L.T, & Alto N.M. (2021). Pathogenic Ubiquitination of GSDMB Inhibits NK-cell Bactericidal Functions.. Cell, 184(12), 3178-3191.e18.

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