HeLa cells and Raw264.7 stable cells were seeded onto glass coverslips. After 24 h, cells were transfected using Lipofectamine 2000 (Invitrogen). Cells were washed 18 h after transfection, fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% BSA for 1 h. Cells were stained with the designated primary antibodies, followed by labelling with anti-mouse Alexa Fluor 568 or 488 and anti-rabbit Alexa Fluor 568 or 488 (Molecular Probes). After antibody labelling, cells were counterstained with Hoechst (33342, Molecular Probes). Cells were imaged with a confocal microscope (Nikon) and were processed with NIS-elements confocal (Nikon).
Nis elements confocal
Manufactured by Nikon
Sourced in Germany
The NIS-Elements Confocal is a software package designed to operate Nikon's line of confocal microscopes. It provides the necessary tools and functionality to control the hardware, acquire images, and perform basic analysis of confocal data.
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2 protocols using nis elements confocal
1
Immunofluorescence Imaging of Transfected Cells
2
Visualizing Yogurt Microstructure with CLSM
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The microstructure of stirred yogurts was visualized by confocal laser scanning microscopy (CLSM). The protein phase was stained with 35 µL of rhodamine B solution (0.016% wt/wt, 479 g mol -1 , Art.-No. 107599, CAS 81-88-9, Merck KGaA, Darmstadt, Germany). Four specimens of each sample were prepared according to Heilig et al. (2009) . Images were taken using a microscope (Eclipse-C1; Nikon GmbH, Düsseldorf, Germany) equipped with a 63× oil immersion objective, controlled via the software NIS-Elements Confocal (Nikon GmbH). Two images were taken at 10°C of each specimen. Finally, the most representative images were selected.
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