For immunostaining, all slides were deparaffinized, rehydrated, and antigen-retrieved using sodium citrate (pH 6.0). The sections were then incubated in blocking buffer consisting of 10% donkey serum and 0.2% Triton X-100 (Sigma-Aldrich, St. Louis, MO, USA) in PBS for 1 hour at room temperature. Subsequently, the slides were incubated with the primary antibody (Alpha SMA, Rabbit anti-Rat, 50-173-6008, Proteintech Group Inc., USA) at a dilution of 1:100 from Cell Signaling Technology (Danvers, MA) overnight at 4°C. After several washes with PBS, the slides were incubated with a fluorescent-labeled secondary antibody [Alexa Fluor 488 Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, PIA32731TR, Invitrogen, USA] for at least 1 hour at room temperature. Following staining, the slides were mounted with a fluorescent-enhanced mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI) (Vector Laboratories, Burlingame, CA, USA). Images were captured using the Ultra viewer ERS dual-spinning disk confocal microscope (PerkinElmer, Waltham MA, USA) equipped with a C-Apochromat (Carl Zeiss, Thornwood, NY, USA) ×10 high dry lens, a C-Apochromat ×40 water immersion lens with a numerical aperture (NA) of 1.2, and an electron multiplier charge-coupled device cooled digital camera (Hamamatsu 12-bit camera, PerkinElmer, Waltham, MA, USA).
Ultra viewer ers dual spinning disk confocal microscope
Manufactured by PerkinElmer
Sourced in United States
The Ultra Viewer ERS dual-spinning disk confocal microscope is a high-performance imaging system designed for advanced fluorescence microscopy applications. It utilizes dual-spinning disk technology to provide high-speed, high-resolution imaging capabilities for a wide range of samples.
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Lab products found in correlation
3 protocols using ultra viewer ers dual spinning disk confocal microscope
1
Immunofluorescence Staining of Alpha SMA
2
Rat CD68 Immunofluorescence Staining
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All slides were deparaffinized, rehydrated, and antigen retrieved (sodium citrate, pH 6.0). After that, the sections were incubated in blocking buffer (10% donkey serum, 0.2% Triton X-100 [Sigma-Aldrich, St. Louis, MO, USA] in PBS) for 1 h at room temperature, followed by incubation with primary antibody (CD68, PIMA513324, Mouse anti-Rat, CD68 Monoclonal Antibody (KP1), Invitrogen™, USA) at 4 °C overnight. After several phosphate-buffered saline (PBS) washes, slides were incubated for at least 1 hr at room temperature in fluorescent-labeled secondary antibody (DyLight™ 594, Goat anti-Mouse IgG (H + L) Secondary Antibody, PI35510, Invitrogen™, USA). After staining, the slides were mounted with a fluorescent-enhanced mounting medium with 4′,6-diamidino-2-phenylindole (DAPI) (Vector Laboratory, Burlingame, CA, USA). Images were taken using the Ultra viewer ERS dual-spinning disk confocal microscope (PerkinElmer, Waltham MA, USA) equipped with a C-Apochromat (Carl Zeiss, Thornwood, NY, USA) ×10 high dry lens, a C-Apochromat ×40 water immersion lens NA 1.2, an electron multiplier charge-coupled device cooled digital camera (Hamamatsu Orce_ERCC 12-bit camera]; PerkinElmer, Waltham, MA, USA)
3
Immunostaining for Retinal Cell Markers
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Deparaffinized sections were stained with Hematoxylin and Eosin (Medical Chemical Corporation, 310-787-6800). For IHC, tissue sections were deparaffinized at 60 °C for 30–45 min and rehydrated followed by heat-induced antigen retrieval for three minutes. Sections were blocked using 5% bovine serum albumin and 0.1% Triton X-100 diluted in PBS for 30 min at room temperature and immunostained overnight with primary antibodies against TRA-1-85 (1:100; MAB3195, R&D Systems), RPE65 (1:500; AB105366, Abcam) and Rhodopsin (1:400; AB3267, Abcam) at 4 °C. Sections were stained with secondary antibodies goat anti-mouse IgG conjugated with rhodamine (1:100; 115 025 146, Jackson Immunoresearch) and goat anti-rabbit IgG conjugated with FITC (1:00; 115 095 144, Jackson Immunoresearch) for one hour at room temperature. Sections were mounted with fluorescein-enhancing mounting medium with DAPI (Vector Laboratory) and imaged using an Ultraviewer ERS dual spinning disk confocal microscope (PerkinElmer) equipped with a C-Apochromat 10 × high dry lens, a C-Apochromat 40 × water immersion lens NA 1.2 and an electron multiplier cooled digital camera (CCD). Images were captured and processed using PerkinElmer Volocity imaging software.
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