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Spots were determined by using a local fluorescence filter.
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Puncta were determined by using a local ratio of the fluorescence image filtered with Gaussian filters (standard deviations of 3.5 and 6.5 pixels) and a ratio threshold of 1.65.
Instead the Cy3 fluorescence image was inverted and filtered using a local equalization filter with a wide pixel window.
Use a local neighborhood watch program.
To visualize the intracellular localization, treated cells were also examined for FITC fluorescence using a multiphoton fluorescence microscope (Bio-Rad, Hercules, CA, USA).
(B) Fluorescence image of the same cells using a fluorescence filter set for green fluorescent protein.
Images were acquired using a BZ-9000 fluorescence microscope (Keyence).
Images were captured using a Leica fluorescence microscope (DM2500, Mannheim, Germany).
Fluorescence was measured using a VersaFluorTM fluorescence spectrophotometer (BioRad).
mCherry fluorescence of bacterial cells suspension was measured every hour using a Luminar fluorescence spectrometer.
Researchers simply add the nanoparticles to living cells and watch for tell-tale color changes using a special fluorescence microscope.
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