Limitations of fluorescence labeling for quantification

<p>The crucial limitation of fluorescence labeling for quantification purposes is that it is almost impossible to have a 100% efficient process. </p> <p>If we want to quantify the number of a given antibody on the surface of a membrane, we could attempt to do single-molecule fluorescence detection, and assuming each antibody is uniquely labelled. </p> <p>This is the challenge which is pervasive in the use of <a class="wikilink" href="/flow_cytometry/">Flow Cytometry</a> to determine <a class="wikilink" href="/loading_ratio_of_nanoparticles/">loading ratio of nanoparticles</a> (see: <a class="wikilink" href="/literature/202111301443_determining_drug_loading_and_release_kinetics_in_nanoparticles/">202111301443 determining drug loading and release kinetics in nanoparticles</a>) or to properly identify particles with low numbers of labeling sites. </p> <p>Another problem is the separating the signal from the background generated by the <a class="wikilink" href="/fluorescence_arising_from_dye_aggregates/">fluorescence arising from dye aggregates</a>.</p>