Fluorescence imaging is an invaluable tool to investigate biomolecular dynamics, mechanics, and interactions in aqueous environments. Two-dimensional materials offer large-area, atomically smooth surfaces for wide-field biomolecule imaging. Despite the success of graphene for on-chip biosensing and biomolecule manipulation, its strong fluorescence-quenching properties pose a challenge for biomolecular investigations that are based on direct optical readouts. Here, we employ few-layer hexagonal boron nitride (hBN) as a precisely tailorable fluorescence spacer between labelled lipid membranes and graphene substrates. By stacking high-quality hBN crystals in the 10–20 nm thickness range on monolayer graphene, we observe distance-dependent fluorescence intensity variations. Remarkably, with hBN spacers as thin as 20 nm, the fluorescence intensity is comparable to bare SiO₂/Si substrates, while the intensity was reduced to 60 % and 80 % with ~10 nm and ~16 nm hBN thicknesses respectively. We confirm that pre-determined hBN thicknesses can be employed to control the non-radiative energy transfer properties of graphene, with fluorescence quenching following a d⁻⁴ distance-dependent behaviour. This seamless integration of electronically active and dielectric van der Waals materials into vertical heterostructures enables multifunctional platforms addressing the manipulation, localization, and visualization of biomolecules for fundamental biophysics and biosensing applications.

Hexagonal boron nitride is a 2D material whose single-layer allotrope has not been intensively studied despite being the substrate for graphene electronics. Its transparency and stronger interlayer adhesion with respect to graphene makes it difficult to work with, and few applications have been proposed. We have developed a transfer technique for this extra-adhesive material that does not require its visual localization, and fabricated mechanical resonators made out of chemical vapor-deposited single-layer hexagonal boron nitride. The suspended material was initially contaminated with polymer residues from the transfer, and the devices showed an unexpected tensioning when cooling them to 3 K. After cleaning in harsh environments with air at 450 °C and ozone, the temperature dependence changed with f₀Q products reaching 2 × 10¹⁰ Hz at room temperature. This work paves the way to the realization of highly sensitive mechanical systems based on hexagonal boron nitride, which could be used as an alternative material to SiN for optomechanics experiments at room temperature.