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Combining ultrastructure expansion microscopy with immunofluorescence and Oligopaint DNA FISH.

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Τίτλος: Combining ultrastructure expansion microscopy with immunofluorescence and Oligopaint DNA FISH.
Συγγραφείς: Raab LM; Unit on Chromosome Dynamics, Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 20892, USA.; Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892, USA., Hockens CB; Unit on Chromosome Dynamics, Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 20892, USA., Lee LS; Unit on Chromosome Dynamics, Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 20892, USA., Runyan RK; Unit on Chromosome Dynamics, Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 20892, USA., Burns EE; Cell and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, 20892, USA., Rusan NM; Cell and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, 20892, USA., Rosin LF; Unit on Chromosome Dynamics, Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 20892, USA. leah.rosin@nih.gov.
Πηγή: Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology [Chromosome Res] 2026 May 21; Vol. 34 (1). Date of Electronic Publication: 2026 May 21.
Τύπος έκδοσης: Journal Article
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Kluwer Academic Publishers Country of Publication: Netherlands NLM ID: 9313452 Publication Model: Electronic Cited Medium: Internet ISSN: 1573-6849 (Electronic) Linking ISSN: 09673849 NLM ISO Abbreviation: Chromosome Res Subsets: MEDLINE
Imprint Name(s): Publication: Dordrecht : Kluwer Academic Publishers
Original Publication: Oxford, UK : Rapid Communications of Oxford, c1993-
Ιατρικοί όροι (MeSH): In Situ Hybridization, Fluorescence*/methods , Fluorescent Antibody Technique*/methods, Ovary/ultrastructure ; DNA/genetics ; Chromatin/ultrastructure ; Drosophila/genetics ; Microscopy, Fluorescence/methods ; Animals ; Female
Περίληψη: Expansion microscopy (ExM) enlarges biological samples by embedding them in a swellable hydrogel, enabling nanoscale imaging of subcellular structures with standard light microscopes. This offers an accessible alternative to super-resolution methods. ExM has yet to be combined with the Oligopaint DNA fluorescence in situ hybridization (FISH) technology. We present an optimized ExM workflow for simultaneous Oligopaint DNA FISH and immunofluorescence (IF) in intact Drosophila ovaries. The protocol incorporates nucleic-acid anchoring, reliable protein retention, and digestion conditions that preserve chromatin while maintaining probe accessibility. Our approach achieves ~ 5 × expansion and strong signal retention, enabling high-resolution studies of nuclear organization.
(© 2026. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)
Competing Interests: Declarations. Competing interests: The authors declare no competing interests.
Σχόλια: Erratum in: Chromosome Res. 2026 Jul 16;34(1):13. doi: 10.1007/s10577-026-09805-6.. (PMID: 42458193)
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Grant Information: ZIA HD009019 United States ImNIH Intramural NIH HHS; ZIA HL006126 United States ImNIH Intramural NIH HHS; 1ZIAHD009019 Division of Intramural Research, NICHD; 1ZIAHL006126 Division of Intramural Research, NHLBI
Contributed Indexing: Keywords: ExM; Expansion microscopy; Meiosis; Oligopaints; Synaptonemal complex
Substance Nomenclature: 9007-49-2 (DNA)
0 (Chromatin)
Entry Date(s): Date Created: 20260520 Date Completed: 20260717 Latest Revision: 20260717
Update Code: 20260717
PubMed Central ID: PMC13220437
DOI: 10.1007/s10577-026-09802-9
PMID: 42162320
Βάση Δεδομένων: MEDLINE
Περιγραφή
ISSN:1573-6849
DOI:10.1007/s10577-026-09802-9