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Finally, the backward-directed SHG was collected in an epi-fluorescence configuration, using the same objective for focusing and for collection.

Nicotinamide adenine dinucleotide (NADH) fluorescence from the same area was measured by switching the microscope to an epi-fluorescence configuration using a 100-W mercury arc lamp source, a 365BP25nm excitation filter, a 450BP65 emission filter, and a 400CLP02 dichroic mirror (NADH-specific XF06 filter unit; Omega Optical, Inc., Brattleboro, VT, USA).

A model immunoassay experiment demonstrates that the reported approach enables increasing signal to noise ratio, resulting in about one order of magnitude improved limit of detection (LOD) with respect to regular total internal reflection fluorescence (TIRF) configuration.

The responsiveness to INH is more complex, and when cells are grown in the absence of Tween, INH addition does not significantly inhibit fluorescence in any configuration that we have tested.

Therefore, we treated CHO-K1 cells stably expressing a GPI-anchored GFP with wortmannin and recorded the fluorescence in TIRF configuration over 150 min.

In this configuration, the fluorescence signal was strongly dominated by that of Zn, owing to the use of zinc white as a primary pigment in the palette of both the upper and lower paintings.

By 6 dpi, the fluorescence of the trans configuration (separate binary vectors carrying FECT/GFP and 35S/p19) had increased so as to be somewhat superior to the cis configuration.

Genetic manipulations carried out in a microwell format allow for the well-by-well analysis using biochemical assays, the sensitivity of which needed to be achieved in a microarray configuration by fluorescence imaging.

For this purpose, CHO-K1 cells expressing the plasma membrane-localized proteins CD147-GFP, GPI-anchored GFP or EGFR-GFP (Weghuber et al., 2011; Lanzerstorfer et al., 2014) were treated with insulin and the respective fluorescence signal in TIRF configuration was recorded within 1 h post-stimulation.

Through analyzing the transition configuration of the fluorescence, we found that the calculated fluorescence has been just the reverse processed of the lowest lying absorption.

This paper investigates plasmonic amplification in two commonly used optical configurations for fluorescence readout of bioassays – epifluorescence (EPF) and total internal reflection fluorescence (TIRF).