Fluorescence in situ hybridization on human metaphase chromosomes detected by near‐field scanning optical microscopy

Fluorescence in situ hybridization on human metaphase chromosomes is detected by near‐field scanning optical microscopy. This combination of cytochemical and scanning probe techniques enables the localization and identification of several fluorescently labelled genomic DNA fragments on a single chromosome with an unprecedented resolution. Three nucleic acid probes are used: pUC1. 77. p1–79 and the plasmid probe α‐spectrin. The hybridization signals are very well resolved in the near‐field fluorescence images, while the exact location of the probes can be correlated accurately with the chromosome topography as afforded by the shear force image.

[1]  B. Stollar,et al.  High resolution detection of DNA–RNA hybrids in situ by indirect immunofluorescence , 1977, Nature.

[2]  B. Hamkalo,et al.  In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold , 1982, The Journal of cell biology.

[3]  R. Toledo-Crow,et al.  Near‐field differential scanning optical microscope with atomic force regulation , 1992 .

[4]  Ambrose,et al.  Single molecule detection and photochemistry on a surface using near-field optical excitation. , 1994, Physical review letters.

[5]  E. Betzig,et al.  Near-Field Spectroscopy of the Quantum Constituents of a Luminescent System , 1994, Science.

[6]  E. Betzig,et al.  Near-Field Optics: Microscopy, Spectroscopy, and Surface Modification Beyond the Diffraction Limit , 1992, Science.

[7]  E. Schuuring,et al.  Multiple and sensitive fluorescence in situ hybridization with rhodamine-, fluorescein-, and coumarin-labeled DNAs. , 1993, Cytogenetics and cell genetics.

[8]  F. Amaldi,et al.  Autoradiographic Detection of Molecular Hybrids between rRNA and DNA in Tissue Sections , 1970, Nature.

[9]  J. Hindley,et al.  Cloning of human satellite III DNA: different components are on different chromosomes. , 1979, Nucleic acids research.

[10]  G van den Engh,et al.  The proximity of DNA sequences in interphase cell nuclei is correlated to genomic distance and permits ordering of cosmids spanning 250 kilobase pairs. , 1989, Genomics.

[11]  D. Ward,et al.  Is non-isotopic in situ hybridization finally coming of age? , 1990, Nature.

[12]  J. Gall,et al.  Formation and detection of RNA-DNA hybrid molecules in cytological preparations. , 1969, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Raouf Fetni,et al.  Detection of small, single-copy genes on protein-G-banded chromosomes by electron microscopy. , 1992, Cytogenetics and cell genetics.

[14]  B. Hamkalo,et al.  Localization of nucleic acid sequences by EM in situ hybridization using colloidal gold labels. , 1989, The American journal of anatomy.

[15]  G Hermanson,et al.  High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. , 1990, Science.

[16]  E. Betzig,et al.  Combined shear force and near‐field scanning optical microscopy , 1992 .

[17]  J. Wiegant,et al.  High-resolution in situ hybridization using DNA halo preparations. , 1992, Human molecular genetics.

[18]  B. D. de Grooth,et al.  Detection of in situ hybridization to human chromosomes with the atomic force microscope. , 1993, Cytometry.

[19]  J. Wiegant,et al.  Multiple colors by fluorescence in situ hybridization using ratio-labelled DNA probes create a molecular karyotype. , 1992, Human molecular genetics.

[20]  J. Wiegant,et al.  A new method for fluorescence microscopical localization of specific DNA sequences by in situ hybridization of fluorochromelabelled RNA. , 1980, Experimental cell research.

[21]  M. Birnstiel,et al.  RNA-DNA Hybrids at the Cytological Level , 1969, Nature.

[22]  R. Drouin,et al.  Simultaneous visualization of chromosome bands and hybridization signal using colloidal-gold labeling in electron microscopy. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[23]  N. F. Hulst,et al.  Poly(diacetylene) Monolayers Studied with a Fluorescence Scanning Near-Field Optical Microscope , 1994 .

[24]  Robert J. Chichester,et al.  Single Molecules Observed by Near-Field Scanning Optical Microscopy , 1993, Science.

[25]  N. F. Hulst,et al.  Optical contrast in near-field techniques , 1995 .

[26]  N. Galjart,et al.  The gene encoding human protective protein (PPGB) is on chromosome 20. , 1991, Genomics.

[27]  T. Ried,et al.  In situ hybridization with fluoresceinated DNA. , 1991, Nucleic acids research.

[28]  A. H. Klahn,et al.  References and Notes , 2022 .

[29]  E. Betzig,et al.  Near-field spectroscopy of single molecules at room temperature , 1994, Nature.