Quantitative three-dimensional evaluation of immunofluorescence staining for large whole mount spheroids with light sheet microscopy
نویسندگان
چکیده
منابع مشابه
A novel protocol of whole mount electro-immunofluorescence staining
PURPOSE To develop a new method of whole mount immunostaining that improves the penetration of staining reagents into the cornea and decreases non-specific binding and background. METHODS Adult mouse corneas were fixed overnight in 4% paraformaldehyde or a mixture of 4% paraformaldehyde and 0.2% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4, at 4 degrees C. After washing with 0.1% Triton X...
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This protocol comprises the entire process of immunofluorescence staining on mouse cochlea whole mount, starting from tissue preparation to the mounting of the tissue. This technique provides "three-dimensional" views of the stained components in order to determine the localization of a protein of interest in the tissue in its natural state and environment.
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We developed a multiple light-sheet microscopy (MLSM) system capable of 3D fluorescence imaging. Employing spatial filter in the excitation arm of a SPIM system, we successfully generated multiple light-sheets. This improves upon the existing SPIM system and is capable of 3D volume imaging by simultaneously illuminating multiple planes in the sample. Theta detection geometry is employed for dat...
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We describe a method for the three-dimensional live imaging of filamentous fungi with light sheet-based fluorescence microscopy (LSFM). LSFM provides completely new opportunities to investigate the biology of fungal cells and other microorganisms with high spatial and temporal resolution. As an example, we study the established aging model Podospora anserina. The protocol explains the mounting ...
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Aberrations, scattering and absorption degrade the performance light-sheet fluorescence microscopes (LSFM). An adaptive optics system to correct for these artefacts and to optimize the light-sheet illumination is presented. This system allows a higher axial resolution to be recovered over the field-of-view of the detection objective. It is standard selective plane illumination microscope (SPIM)...
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ژورنال
عنوان ژورنال: Biomedical Optics Express
سال: 2017
ISSN: 2156-7085,2156-7085
DOI: 10.1364/boe.8.000484