VCaP cells were fixed with 3.7% paraformaldehyde, and then permeabilized with 0.1% (w/v) saponin for 15 min. Cells were co-incubated with primary antibodies against AR and menin or ASH2L for 12hr at 4°C, followed by incubating with appropriate Alexa-Fluor-conjugated secondary antibodies for 30 min at 37°C. Cells were washed and mounted onto glass slides using VectaShield mounting medium containing DAPI. Samples were analyzed using a Nikon A1 laser-scanning confocal microscope equipped with a Plan-Apo ×63/1.4 numerical aperture oil lens objective. Acquired images were then analyzed using ImageJ software (version 1.41o). Scale bar: 10μm.
Plan apo 63 1.4 numerical aperture oil lens objective
Manufactured by Nikon
The Plan-Apo ×63/1.4 numerical aperture oil lens objective is a high-performance microscope objective lens designed for advanced microscopy applications. It features a 63x magnification and a numerical aperture of 1.4, which delivers exceptional image resolution and clarity. The lens is optimized for use with immersion oil, providing enhanced optical performance.
Automatically generated - may contain errors
4 protocols using plan apo 63 1.4 numerical aperture oil lens objective
1
Immunofluorescence Imaging of AR and Chromatin Regulators
2
Immunofluorescence Imaging of EED, BMI1, and RING1B
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DU145 or LNCaP cells were fixed with 3.7% paraformaldehyde, and then permeabilized with 0.1% (w/v) saponin (Sigma) for 15 min. Cells were co-incubated with primary antibodies against EED and BMI1 or EED and RING1B (1:100 dilution) for 12hr at 4 °C, followed by incubating with appropriate Alexa-Fluor-conjugated secondary antibodies (1:200 dilution) for 30 min at 37°C. Cells were washed and mounted onto glass slides using Vectashield mounting medium (Vector Laboratories, Burlingame, CA) containing DAPI. Samples were analyzed using a Nikon A1 laser-scanning confocal microscope equipped with a Plan-Apo ×63/1.4 numerical aperture oil lens objective. Acquired images were then analyzed using ImageJ software (version 1.41). Co-localization was analyzed in Image J software using JACoP plugin. 15–20 nuclei were counted and co-localization was measured using the Manders coefficient to evaluate the overlap in fluorescence.
3
Immunofluorescence Imaging of EED, BMI1, and RING1B
4
Immunofluorescence Imaging of AR and Chromatin Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VCaP cells were fixed with 3.7% paraformaldehyde, and then permeabilized with 0.1% (w/v) saponin for 15 min. Cells were co-incubated with primary antibodies against AR and menin or ASH2L for 12hr at 4°C, followed by incubating with appropriate Alexa-Fluor-conjugated secondary antibodies for 30 min at 37°C. Cells were washed and mounted onto glass slides using VectaShield mounting medium containing DAPI. Samples were analyzed using a Nikon A1 laser-scanning confocal microscope equipped with a Plan-Apo ×63/1.4 numerical aperture oil lens objective. Acquired images were then analyzed using ImageJ software (version 1.41o). Scale bar: 10μm.
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