Frozen sections were fixed for 20 min in 4% paraformaldheyde, washed with PBS, and blocked with 2% BSA (Sigma-Aldrich) in PBS for 30 min. Unspecific binding sites were saturated with 5% goat serum, 2% BSA, and 0.1% glycine in PBS for 1 h. FITC-conjugated anti-CD41 was diluted 1:100, anti-FN 1:500, and anti–type III collagen 1:100 and incubated overnight at 4°C in washing buffer (0.2% BSA, 0.1% Tween 20 in PBS). 594 Alexa Fluor–conjugated secondary antibodies were purchased from Invitrogen. Nuclei were counterstained using Hoechst 33258 (100 ng/ml in PBS) at room temperature. Sections were then mounted with micro-cover glasses using Fluoro-mount (Bio-Optica). Negative controls were routinely performed by omitting the primary antibodies. Images were acquired using the Olympus BX51 fluorescence microscopy (Olympus Deutschland) and 10×/0.30 or 20×/0.75 Olympus UplanF1 objectives or with a TCS SP8 confocal laser scanning microscope (Leica).
Uplanf1
Manufactured by Olympus
Sourced in United States
The UplanF1 is a high-quality objective lens designed for use in microscopy applications. It features a flat field of view and high numerical aperture, providing excellent image quality and resolution. The lens is suitable for a variety of sample types and can be used in both brightfield and darkfield microscopy.
Automatically generated - may contain errors
2 protocols using uplanf1
1
Immunofluorescence Staining of Frozen Sections
2
Fungal Isolate Phenotypic Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The phenotypic variant of the fungal isolate was determined by visual inspection. The fungus was grown in PDA broth at 28 °C for 7 days, then the Petri plate was photographed. The hyphae and conidia from the fungal cultures were stained with lactophenol blue solution (LPCB), Sigma-Aldrich, Saint Louis, MO, USA) and observed under a phase-contrast light microscope using an Olympus BX51 with a 60× objective UPlanF1.
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