Sp8 gsted

Manufactured by Leica camera

The Leica SP8 gSTED is a high-performance confocal microscope system designed for advanced fluorescence imaging. It combines the capabilities of confocal microscopy with the super-resolution techniques of gated Stimulated Emission Depletion (gSTED) technology. The SP8 gSTED enables researchers to achieve nanoscale resolution beyond the diffraction limit of conventional light microscopy, providing detailed insights into cellular and subcellular structures.

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Lab products found in correlation

Prolong gold with dapi, by Thermo Fisher Scientific (2 mentions) Chambered coverslip, by Ibidi (2 mentions)

2 protocols using sp8 gsted

Immunofluorescence Localization of IRF8

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HeLa cells were plated on chambered coverslip (#80,826, iBidi) and were left untransfected or were transiently transfected with IRF8-WT or mutant cDNA constructs or with EV. Twenty-four hours later, cells were fixed 15 min in 4% formaldehyde in phosphate-buffered saline (PBS), pH 7.4 at 37 °C. The cells were then incubated overnight at 4 °C with primary antibody (unconjugated IRF8, clone D20D8, #5628, Cell Signaling). They were washed three times in PBS, stained by incubation with secondary antibodies for one hour at room temperature (goat anti-rabbit IgG Alexa Fluor 488 (#A-11034), and left in ProLong Gold with DAPI (#P36931, Thermo Fisher). Cells were then visualized by confocal microscopy (× 63 oil immersion lens, SP8 gSTED, Leica). Images were analyzed using Fiji software.

Rosain J., Bernasconi A., Prieto E., Caputi L., Le Voyer T., Buda G., Marti M., Bohlen J., Neehus A.L., Castaños C., Gallagher R., Dorgham K., Oleastro M., Perez L., Danielian S., Dipierri J.E., Casanova J.L., Bustamante J, & Villa M. (2022). Pulmonary Alveolar Proteinosis and Multiple Infectious Diseases in a Child with Autosomal Recessive Complete IRF8 Deficiency. Journal of Clinical Immunology, 42(5), 975-985.

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IFN-γ Stimulation of SV40-fibroblasts

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SV40-fibroblasts were plated on chambered coverslips (#80826, iBidi). The following day, they were left unstimulated or were stimulated for the indicated times with 10³ IU/mL IFN-γ (Imukin, Horizon Pharma). Cells were fixed by incubation for 15 minutes in 4% formaldehyde in phosphate-buffered saline (PBS), pH 7.4 at 37°C. The cells were then incubated overnight at 4°C with primary antibody (unconjugated, clone D5E4, #8478, Cell Signaling). They were washed three times in PBS, stained by incubation with secondary antibodies for one hour at room temperature (goat anti-rabbit IgG Alexa Fluor 555, #A21429), and left in ProLong Gold with DAPI (#P36931, Thermo Fisher Scientific). Cells were then visualized by confocal microscopy (×63 oil immersion lens, SP8 gSTED, Leica). Images were analyzed with Fiji software.

Rosain J., Neehus A.L., Manry J., Yang R., Le Pen J., Daher W., Liu Z., Chan Y.H., Tahuil N., Türel Ö., Bourgey M., Ogishi M., Doisne J.M., Izquierdo H.M., Shirasaki T., Le Voyer T., Guérin A., Bastard P., Moncada-Velez M., Han J.E., Khan T., Rapaport F., Hong S.H., Cheung A., Haake K., Mindt B.C., Perez L., Philippot Q., Lee D., Zhang P., Rinchai D., Al Ali F., Ata M.M., Rahman M., Peel J.N., Heissel S., Molina H., Kendir-Demirkol Y., Bailey R., Zhao S., Bohlen J., Mancini M., Seeleuthner Y., Roelens M., Lorenzo L., Soudée C., Paz M.E., Gonzalez M.L., Jeljeli M., Soulier J., Romana S., L’Honneur A.S., Materna M., Martínez-Barricarte R., Pochon M., Oleaga-Quintas C., Michev A., Migaud M., Lévy R., Alyanakian M.A., Rozenberg F., Croft C.A., Vogt G., Emile J.F., Kremer L., Ma C.S., Fritz J.H., Lemon S.M., Spaan A.N., Manel N., Abel L., MacDonald M.R., Boisson-Dupuis S., Marr N., Tangye S.G., Di Santo J.P., Zhang Q., Zhang S.Y., Rice C.M., Béziat V., Lachmann N., Langlais D., Casanova J.L., Gros P, & Bustamante J. (2023). Human IRF1 governs macrophagic IFN-γ immunity to mycobacteria. Cell, 186(3), 621-645.e33.

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