IQ-1

Near-infrared (NIR) surface-enhanced resonance Raman (SERRS) nanoprobes have found wide applications in biomedicine; however, almost all of these nanoprobes are fluorescent because the resonant Raman dyes used cannot be fully quenched onto the underlying plasmonic nanoparticles. Therefore, suppressing the fluorescence backgrounds in resonant Raman spectroscopy imaging is extremely important. In this work, we use a black hole quencher, IQ1, as a Raman dye to develop absolutely nonfluorescent NIR resonant SERRS NPs. Ultrafast spectroscopy clarifies that the nonfluorescent mechanism of the dyes is attributed to the ultrafast internal conversion at the subpicosecond scale, which quenches the fluorescence of excited states. The resultant nanoprobes exhibit zero fluorescent background, femtomolar-level sensitivity (100 fM) as well as superb photostability (τ = 10006 s) without fluorescence photobleaching, outperforming that of fluorescent counterparts. More importantly, the SERRS NPs show a synergistic photothermal effect originating from the dye molecule-plasmon interactions, achieving a high photothermal conversion efficiency of 64.94%. Featuring these excellent properties, these SERRS NPs allow for longitudinally photostable cellular imaging and enhanced photothermal elimination of cancer cells. To the best of our knowledge, this is the first example of absolutely nonfluorescent NIR SERRS NPs, opening up promising applications for improved phototheranostics.

Understanding the molecular identity of human pluripotent stem cell (hPSC)-derived cardiac progenitors and mechanisms controlling their proliferation and differentiation is valuable for developmental biology and regenerative medicine.

Here, we show that chemical modulation of histone acetyl transferases (by IQ-1) and WNT (by CHIR99021) synergistically enables the transient and reversible block of directed cardiac differentiation progression on hPSCs. The resulting stabilized cardiovascular progenitors (SCPs) are characterized by ISL1pos/KI-67pos/NKX2-5neg expression. In the presence of the chemical inhibitors, SCPs maintain a proliferation quiescent state. Upon small molecules, removal SCPs resume proliferation and concomitant NKX2-5 up-regulation triggers cell-autonomous differentiation into cardiomyocytes. Directed differentiation of SCPs into the endothelial and smooth muscle lineages confirms their full developmental potential typical of bona fide cardiovascular progenitors. Single-cell RNA-sequencing-based transcriptional profiling of our in vitro generated human SCPs notably reflects the dynamic cellular composition of E8.25-E9.25 posterior second heart field of mouse hearts, hallmarked by nuclear receptor sub-family 2 group F member 2 expression. Investigating molecular mechanisms of SCP stabilization, we found that the cell-autonomously regulated retinoic acid and BMP signalling is governing SCP transition from quiescence towards proliferation and cell-autonomous differentiation, reminiscent of a niche-like behaviour.

The chemically defined and reversible nature of our stabilization approach provides an unprecedented opportunity to dissect mechanisms of cardiovascular progenitors’ specification and reveal their cellular and molecular properties.