Porous Vycor glass (Corning 7930) was purchased from Advanced Glass and Ceramics (Holden, MA). Silicon phthalocyanine dichloride, aluminum(III) phthalocyanine chloride tetrasulfonic acid, 9-bromo-2-methylnon-2-ene, sodium sulfite, triphenyl phosphine (PPh3), benzoic acid, dimethylsulfone, DMF, CH2Cl2, ethanol, D2O, and DMSO-d6 were purchased from commercial suppliers and were used as received. Dichloromethane was distilled over phosphorus pentoxide prior to use. Deionized water was purified with a U.S. Filter Corporation deionization system (Vineland, NJ). Nuclear magnetic resonance (NMR) data were recorded on a Bruker Avance 400 spectrometer operating at 400 MHz for 1H NMR and at 100.6 MHz for 13C NMR. UV–vis data were collected on a Hitachi UV–vis U-2001 instrument. FAB-mass spectrometry data were collected on a JEOL JMS-HX110 spectrometer using a m-nitrobenzyl alcohol matrix, a 10 kV acceleration voltage, and a Xe beam FAB gun (6 kV) on the MS-1 ion source. Infrared spectra were recorded on a Nicolet iS10 FT-IR spectrometer. Solution temperatures were measured with a digital pyrometer (Thermo Scientific). An Olympus FluoView FV10i confocal fluorescence microscope was used to analyze stained E. coli and assess membrane permeability following singlet oxygen exposure.
Fluoview fv10i confocal fluorescence microscope
Manufactured by Olympus
Sourced in Japan
The FluoView FV10i is a confocal fluorescence microscope designed for high-resolution imaging of biological samples. It utilizes laser excitation and advanced optics to generate detailed, three-dimensional images of fluorescently labeled specimens.
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Lab products found in correlation
3 protocols using fluoview fv10i confocal fluorescence microscope
1
Porous Glass Photosensitizer Synthesis
2
Analyzing Nuclear Morphology with DAPI
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Nuclear morphology was observed with the use of DAPI (4′,6-diamidino-2-phenylindole) staining. The cells (1 × 105 cells/well) were seeded onto the cover slip in the six-well culture plate and then pretreated with the LY294002 (5 μM) or vehicle for 2 h prior to the cariporide (160 μM) for 72 h at 37°C. The cover slip for adherence cells were washed with 1× PBS 3 times, fixed in 4% paraformaldehyde at room temperature for 10 min, and washed with 1× PBS. The cover slip was resuspended in the DAPI (3 ng/ml in 1× PBS) for 5 min in the dark and washed with 1× PBS. The cover slip was placed on the slides, and was then mounted using a mounting medium (08381; Polysciences, Inc., USA). The apoptotic cells were observed with the FluoView FV10i confocal fluorescence microscope (Olympus Corporation, Japan).
3
Immunofluorescence Staining of Phosphorylated Proteins
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Cells were grown on coverslips, fixed with 3.7% formaldehyde in PBS for 15 min, washed twice with PBS for 5 min, and then permeabilized with PBS/0.2% Triton X-100 for 10 min. Cells were then incubated twice with PBS/0.01% Triton X-100 for 5 min, blocked with PBS/3% BSA/0.01% Triton X-100 for 1 h, and incubated with polyclonal antibodies against p-p53-Ser15 or p-Histone H2A.X-Ser139 antibodies (Cell Signaling Technology) at 1:1000 in PBS/1% BSA/0.01% Triton X-100. Cells were then washed twice with PBS/0.01% Triton X-100 for 5 min and incubated with Alexa Fluor 568-conjugated goat anti-mouse IgG (Molecular Probes, Eugene, OR, USA) at 1:500 in PBS/1% BSA/0.01% Triton X-100. Cells were mounted with Vectashield Mounting Medium containing 4′,6-diamidino-2-phenylindole (Vector Laboratories) and visualized and photographed using a FLUOVIEW FV10i confocal fluorescence microscope (Olympus, Shinjuku, Japan).
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