For all comparisons, serial sections were stained with the indicated method and the similar serial fields were imaged at indicated times, and under indicated conditions. All pictures were taken using a spectral camera with tunable filter (Nuance FX; Perkin Elmer) and illumination with a LED at 425nm (Cool-LED PE-2) or 435nm (Cool-LED PE-1000). LED illumination of Qdots was chosen for even, stable illumination without photobleaching of original Qdots [5 (link)].
Quantification of the intensity of fluorescence was performed using the Nuance software, with either autothreshold when different slides were compared or selection of region of interest (ROI) through automatic or manual thresholding and use of the same region to analyse the exact same areas when a time course of the fluorescence intensity was performed (slide kept on microscope stage during the whole of the experiment) or using AxioVision (Zeiss) software.
Quantification of the intensity of fluorescence was performed using the Nuance software, with either autothreshold when different slides were compared or selection of region of interest (ROI) through automatic or manual thresholding and use of the same region to analyse the exact same areas when a time course of the fluorescence intensity was performed (slide kept on microscope stage during the whole of the experiment) or using AxioVision (Zeiss) software.
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