Rhodamine labeled Phaseolus Vulgaris Leucoagglutinin (PHA-L) was purchased from Vector Labs (RL-1112). Lectin staining was performed as previously described (Blackburn and Lupashin, 2016 (link)) with minor modifications. Briefly, cells were cultured as indicated to 70–80% confluency on collagen-coated coverslips. Cells were washed with PBS and fixed in 1% PFA for 15 min, quenched with 50 mM NH4Cl for 10 min, and washed with PBS, followed by a 30 min block with PBSB. The coverslips were incubated in PBSB containing 2 μg/mL PHA-L in a 4°C cold-room (kept in the dark during incubation) with gentle rocking for 30 min. Then, coverslips were stained with Hoechst 33342, washed with PBS three times, and mounted for imaging. The images were obtained using a Nikon NIS-Elements C confocal microscope with a 60× oil lens and Z-stacks at 0.3 μm intervals. Maximum intensity projections were processed and quantified using Nikon NIS-Elements analysis software. All images in the same experiment were captured and processed with the same setting.
Nis elements c confocal microscope
Manufactured by Nikon
The Nikon NIS-Elements C confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It features a confocal scanning mechanism that enables optical sectioning and three-dimensional imaging of samples. The microscope is equipped with a range of imaging modes, including fluorescence, reflection, and transmitted light, allowing for the visualization and analysis of a variety of biological and material samples.
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Lab products found in correlation
2 protocols using nis elements c confocal microscope
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Visualization and Quantification of Lectin Binding
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Visualization and Quantification of Lectin Binding
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Rhodamine labeled Phaseolus Vulgaris Leucoagglutinin (PHA-L) was purchased from Vector Labs (RL-1112). Lectin staining was performed as previously described (Blackburn and Lupashin, 2016 (link)) with minor modifications. Briefly, cells were cultured as indicated to 70–80% confluency on collagen-coated coverslips. Cells were washed with PBS and fixed in 1% PFA for 15 min, quenched with 50 mM NH4Cl for 10 min, and washed with PBS, followed by a 30 min block with PBSB. The coverslips were incubated in PBSB containing 2 μg/mL PHA-L in a 4°C cold-room (kept in the dark during incubation) with gentle rocking for 30 min. Then, coverslips were stained with Hoechst 33342, washed with PBS three times, and mounted for imaging. The images were obtained using a Nikon NIS-Elements C confocal microscope with a 60× oil lens and Z-stacks at 0.3 μm intervals. Maximum intensity projections were processed and quantified using Nikon NIS-Elements analysis software. All images in the same experiment were captured and processed with the same setting.
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