To validate the automated quantification of bound and internalized bacteria we compared the results of automated image analysis to human counting of the same images.
Bound and internalized bacteria per cell were manually counted for 50 cells in control and cytochalasin D treated conditions and compared to software quantification of the same 50 cells.
Specific fluorescent-labeled antibodies directed against the opsonizing agent were used to discriminate between bound and internalized particles.
It should be noted that flow cytometry does not differentiate between bound and internalized nanoparticles; however, cell uptake was confirmed by confocal microscopy.
Here, we show that the model correctly predicts the proportions of free, bound, and internalized receptors, as a function of external LDL concentration, assuming the system is at steady-state.
The EIA measures both bound and free forms of VEGF.
Radioactivity of surface-bound and internalized fraction was measured in a WIZARD2 Automatic gamma counter (Perkin Elmer, Waltham, MA, USA).
Subsequently, the amount of free, membrane-bound, and internalized activity were quantified for each sample using a γ-counter.
Confocal microscopy showed membrane-bound and internalized fluorescence in 231-H2N cells after exposure to FITC-F3 for 2 h.
Samples of the membrane-bound fractions and the internalized fractions (lysates) were measured for their radioactivity content using a well-type gamma counter (1470 Wizard, Wallac), and then, the percentage of membrane-bound and internalized fractions was calculated.
