Tools of the TradePublished: 01 October 2026Aiping Wang1,2 na1 &Jie Zhang3,4 na1 Nature Reviews Molecular Cell Biology (2026) Cite this articleSave articleView saved researchHigher-order chromatin architecture is intimately linked to gene expression regulation, cell-fate decisions, and oncogenesis. Achieving nanoscale imaging of chromatin in living biological systems remains technically challenging. Single-molecule localization microscopy (SMLM) methods, such as (direct) stochastic optical reconstruction microscopy ((d)STORM) and MINimal photon FLUXes (MINFLUX), achieve sufficient localization precision to enable the resolution of chromatin nanostructure. However, these methods require photoswitchable fluorophores that can efficiently switch between dark and bright states. Conventional SMLM or DNA dyes require harsh environments or fixation methods or lack the photoswitching properties needed for SMLM, creating an unmet need for live-cell SMLM imaging of chromatin.Compatible with super-resolution imaging systems including STORM and MINFLUX, HoTs enable quantitative chromatin analysis in living cultured cells and tissue sections and are useful for a wide range of applications. In living cells, HoTs enable long-term real-time visualization of chromatin architecture and dynamics. When combined with a MINFLUX platform, they achieve near-nanometre localization precision for two-dimensional (2D) and three-dimensional (3D) visualization of DNA fibres. When combined with OligoSTORM, genomic loci can be visualized within their native chromatin environment. Notably, we also visualized chromatin architecture in HoT-labelled human paraffin-embedded colon sections. This allowed direct nanoscale visualization of nuclear architecture and chromatin compaction in both normal and cancerous tissues. These properties will establish HoT probes as a powerful tool for basic chromatin biology with potential applications in clinical diagnostics.This is a preview of subscription content, access via your institutionAccess options Access through your institutionAccess Nature and 54 other Nature Portfolio journalsGet Nature+, our best-value online-access subscription27,99 € / 30 dayscancel any timeLearn moreSubscribe to this journalReceive 12 print issues and online access269,00 € per yearonly 22,42 € per issueLearn moreBuy this articlePurchase on SpringerLinkInstant access to the full article PDF.39,95 €Prices may be subject to local taxes which are calculated during checkoutSubjectsCell biologyMolecular biologyReferencesOriginal articleWang, A. et al. HoT auto-blinking probes enable real-time, super-resolution chromatin imaging in live cells and tissues. Mol. Cell 86, 3840–3855.e8 https://doi.org/10.1016/j.molcel.2026.08.010 (2026)Article PubMed Google Scholar Download referencesAuthor informationAuthor notesThese authors contributed equally: Aiping Wang, Jie Zhang.Authors and AffiliationsUniversitat Pompeu Fabra (UPF), Barcelona, SpainAiping WangMedical Research Institute, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, ChinaAiping WangDepartment of Chemistry and COSDAF (Centre of Super-Diamond and Advanced Films), City University of Hong Kong, Hong Kong, ChinaJie ZhangInstitute of Digital Medicine (IDM), City University of Hong Kong, Hong Kong, ChinaJie ZhangAuthorsAiping WangView author publicationsSearch author on:PubMed Google ScholarJie ZhangView author publicationsSearch author on:PubMed Google ScholarCorresponding authorCorrespondence to Aiping Wang.Ethics declarationsCompeting interestsJ.Z. and A.W. have filed a patent application related to this work.Rights and permissionsReprints and permissionsAbout this article