The use of nanoparticles
as carriers to deliver pharmacologically
active compounds to specific parts of the body via the bloodstream
is a promising therapeutic approach for the effective treatment of
various diseases. To reach their target sites, nanocarriers (NCs)
need to circulate in the bloodstream for prolonged periods without
aggregation, degradation, or cargo loss. However, it is very difficult
to identify and monitor small-sized NCs and their cargo in the dense
and highly complex blood environment. Here, we present a new fluorescence
correlation spectroscopy-based method that allows the precise characterization
of fluorescently labeled NCs in samples of less than 50 μL of
whole blood. The NC size, concentration, and loading efficiency can
be measured to evaluate circulation times, stability, or premature
drug release. We apply the new method to follow the fate of pH-degradable
fluorescent cargo-loaded nanogels in the blood of live mice for periods
of up to 72 h.