Reliable and reproducible measurement of spatial resolution is essential for validating and comparing the performance of two-photon microscopy systems. We present fluorescent nanodiamonds as robust, photostable, reusable, and biocompatible probes for benchmarking spatial resolution. Owing to their nanoscale dimensions and stable fluorescence, nanodiamonds act as near-ideal point-like emitters, enabling accurate characterisation of the point spread function across varying imaging conditions. We demonstrate that nanodiamond-based phantoms serve as a reliable alternative to conventional fluorescent beads embedded in agarose, while at the same time offering advantages in stability and optical properties. Our results position nanodiamond phantoms as a next-generation calibration material that bridges ease of use and reproducibility, advancing quantitative imaging and cross-platform comparability in modern fluorescence microscopy. ### Competing Interest Statement The authors have declared no competing interest. Wellcome Trust, https://ror.org/029chgv08, Investigator Award in Science 220277/Z20/Z European Research Council, https://ror.org/0472cxd90, ERC-StG “NeuroVisEco” 677687, ERC AdG “Cones4Action” covered under the UK’s EPSRC guarantee scheme EP/Z533981/1 UK Research and Innovation, https://ror.org/001aqnf71, BBSRC, BB/R014817/1, BBSRC, BB/W013509/1 International Human Frontier Science Program Organization, https://ror.org/02ebx7v45, RGP001/2025 Leverhulme Trust, https://ror.org/012mzw131, PLP-2017-005, RPG-2021-026, RPG-2-23-042 Lister Institute of Preventive Medicine Gdańsk University of Technology, https://ror.org/006x4sc24, Nobelium DEC-4/1/2024/IDUB/I.1a/No, Platinum 1/1/2025/IDUB/I.1B/Pt

Nuclear hyperpolarization of 13C spins is performed at ambient conditions in micro- and nanodiamonds.

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Spin-enhanced lateral flow tests use nanodiamonds for the sensitive, robust detection of disease biomarkers. Here, authors report a clinical evaluation of a test for SARS-CoV-2 antigen, finding 95.1% sensitivity (Ct ≤ 30) and 100% specificity, with detection 2.0 days earlier than conventional tests.

Flawless nanodiamond synthesis that wasn't though possible accomplished by transforming adamantane

Nanodiamonds are tiny diamond particles that usually measure mere millionths of a millimeter. They are extremely hard, stable and heat-resistant and highly adaptable for various purposes in medicine, new materials, catalysis and energy technology.

Nature - Bottom-Up Synthesis of Molecular Nanodiamond from Nanographene

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In recent years, the field of quantum sensing has emerged as one of the most exciting frontiers in modern science and technology. At the…

An international research team led by UCL, Great Ormond Street Hospital, and KU Leuven is using 3D-printing and nanodiamonds to design treatments that could help babies repair their damaged lungs whil...

Nanothermometry in living cells is crucial for understanding heat diffusion and advancing cellular biology and drug discovery. Previous studies have reported controversial intracellular temperature v...