Lesion areas are distinguished from normal tissues surrounding
them by distinct physiological characteristics. These features serve
as biological hallmarks with which targeted biomedical imaging of
the lesion sites can be achieved. Although tremendous efforts have
been devoted to providing smart imaging probes with the capability
of visualizing the physiological hallmarks at the molecular level,
the majority of them are merely able to derive anatomical information
from the tissues of interest, and thus are not suitable for taking
part in in vivo quantification of the biomarkers.
Recent advances in chemical construction of advanced ratiometric nanoprobes
(RNPs) have enabled a horizon for quantitatively monitoring the biological
abnormalities in vivo. In contrast to the conventional
probes whose dependency of output on single-signal profiles restricts
them from taking part in quantitative practices, RNPs are designed
to provide information in two channels, affording a self-calibration
opportunity to exclude the analyte-independent factors from the outputs
and address the issue. Most of the conventional RNPs have encountered
several challenges regarding the reliability and sufficiency of the
obtained data for high-performance imaging. In this Review, we have
summarized the recent progresses in developing highly advanced RNPs
with the capabilities of deriving maximized information from the lesion
areas of interest as well as adapting themselves to the complex biological
systems in order to minimize microenvironmental-induced falsified
signals. To provide a better outlook on the current advanced RNPs,
nanoprobes based on optical, photoacoustic, and magnetic resonance
imaging modalities for visualizing a wide range of analytes such as
pH, reactive species, and different derivations of amino acids have
been included. Furthermore, the physicochemical properties of the
RNPs, the major constituents of the nanosystems and the analyte recognition
mechanisms have been introduced. Moreover, the alterations in the
values of the ratiometric signal in response to the analyte of interest
as well as the time at which the highest value is achieved, have been
included for most of RNPs discussed in this Review. Finally, the challenges
as well as future perspectives in the field are discussed.