Fluorescent Protein Analysis with DAPI Staining

Fluorescently tagged proteins and DAPI (4′, 6-diamidino-2-phenylindole) staining for nuclear analysis were visualized as described in [74 (link)]. A DM6000 microscope (Leica) equipped with a 100×/1.40 NA (numerical aperture) oil immersion objective and a DFC350 FX digital charge-coupled device camera (Leica) was used to image the cells. The obtained images were processed and analyzed with LAS AF (Leica) and ImageJ (http://rsbweb.nih.gov/ij/) software.

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de Oya I.G., Manzano-López J., Álvarez-Llamas A., Vázquez-Aroca M.D., Cepeda-García C, & Monje-Casas F. (2023). Characterization of a novel interaction of the Nup159 nucleoporin with asymmetrically localized spindle pole body proteins and its link with autophagy. PLOS Biology, 21(8), e3002224.

Publication 2023

DM6000 microscope DFC350 FX LAS AF ImageJ

Camera Cells Dapi Device Immersion Microscope Proteins

Corresponding Organization : Universidad de Sevilla

Top 5 similar protocols

Variable analysis

independent variables

Fluorescently tagged proteins

dependent variables

Visualization of fluorescently tagged proteins
Nuclear analysis

control variables

DAPI (4′, 6-diamidino-2-phenylindole) staining for nuclear analysis
DM6000 microscope (Leica) equipped with a 100×/1.40 NA (numerical aperture) oil immersion objective
DFC350 FX digital charge-coupled device camera (Leica)
LAS AF (Leica) and ImageJ software for image processing and analysis

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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