Literature DB >> 1738608

Sensitive detection of RNAs in single cells by flow cytometry.

H Yu1, L Ernst, M Wagner, A Waggoner.   

Abstract

A rapid and sensitive fluorescent in situ hybridization method has been developed to probe RNA contents of individual cells by flow cytometry. Fixed cells in suspension were hybridized with 5' end-fluorophore-labeled oligodeoxynucleotides complementary to defined regions of the RNA of interest and analyzed by flow cytometry. With this method, we monitored combinations of histone H4 mRNA, 18S rRNA and 28S rRNA levels in synchronized HeLa S3 cells by multicolor analysis. A fluorescence signal equivalent to 1800 copies of histone H4 mRNA per cell was detected with signal-to-background ratio of 5.4. If non-specific binding of the fluorophore-labeled probe can be reduced, as few as 100 copies of mRNA of the size of H4 could be detected in individual cells by flow cytometry.

Entities:  

Mesh：

Substances：

Year:  1992        PMID： 1738608      PMCID： PMC310329          DOI： 10.1093/nar/20.1.83

Source DB:  PubMed          Journal:  Nucleic Acids Res        ISSN： 0305-1048            Impact factor:   16.971


  25 in total

1.  Mapping of single-copy DNA sequences on human chromosomes by in situ hybridization with biotinylated probes: enhancement of detection sensitivity by intensified-fluorescence digital-imaging microscopy.

Authors:  E Viegas-Pequignot; B Dutrillaux; H Magdelenat; M Coppey-Moisan
Journal:  Proc Natl Acad Sci U S A       Date:  1989-01       Impact factor: 11.205

2.  Comparison of 35S and biotin as labels for in situ hybridization: use of an HPV model system.

Authors:  E R Unger; M L Hammer; M L Chenggis
Journal:  J Histochem Cytochem       Date:  1991-01       Impact factor: 2.479

3.  Detection of DNA sequences in nuclei in suspension by in situ hybridization and dual beam flow cytometry.

Authors:  B Trask; G van den Engh; J Landegent; N J in de Wal; M van der Ploeg
Journal:  Science       Date:  1985-12-20       Impact factor: 47.728

4.  High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones.

Authors:  P Lichter; C J Tang; K Call; G Hermanson; G A Evans; D Housman; D C Ward
Journal:  Science       Date:  1990-01-05       Impact factor: 47.728

5.  Highly localized tracks of specific transcripts within interphase nuclei visualized by in situ hybridization.

Authors:  J B Lawrence; R H Singer; L M Marselle
Journal:  Cell       Date:  1989-05-05       Impact factor: 41.582

6.  Five-parameter fluorescence imaging: wound healing of living Swiss 3T3 cells.

Authors:  R DeBiasio; G R Bright; L A Ernst; A S Waggoner; D L Taylor
Journal:  J Cell Biol       Date:  1987-10       Impact factor: 10.539

7.  The proximity of DNA sequences in interphase cell nuclei is correlated to genomic distance and permits ordering of cosmids spanning 250 kilobase pairs.

Authors:  B Trask; D Pinkel; G van den Engh
Journal:  Genomics       Date:  1989-11       Impact factor: 5.736

8.  Reassessment of histone gene expression during cell cycle in human cells by using homologous H4 histone cDNA.

Authors:  S Detke; A Lichtler; I Phillips; J Stein; G Stein
Journal:  Proc Natl Acad Sci U S A       Date:  1979-10       Impact factor: 11.205

9.  Flow cytometric detection of beta-globin mRNA in murine haemopoietic tissues using fluorescent in situ hybridization.

Authors:  J A Bayer; J G Bauman
Journal:  Cytometry       Date:  1990

10.  Sensitive, high-resolution chromatin and chromosome mapping in situ: presence and orientation of two closely integrated copies of EBV in a lymphoma line.

Authors:  J B Lawrence; C A Villnave; R H Singer
Journal:  Cell       Date:  1988-01-15       Impact factor: 41.582
View more
  10 in total

1.  Myosin II transport, organization, and phosphorylation: evidence for cortical flow/solation-contraction coupling during cytokinesis and cell locomotion.

Authors:  R L DeBiasio; G M LaRocca; P L Post; D L Taylor
Journal:  Mol Biol Cell       Date:  1996-08       Impact factor: 4.138

Review 2.  RNA molecules lighting up under the microscope.

Authors:  R W Dirks
Journal:  Histochem Cell Biol       Date:  1996-08       Impact factor: 4.304

3.  Directly labeled DNA probes using fluorescent nucleotides with different length linkers.

Authors:  Z Zhu; J Chao; H Yu; A S Waggoner
Journal:  Nucleic Acids Res       Date:  1994-08-25       Impact factor: 16.971

4.  Incorporation of reporter molecule-labeled nucleotides by DNA polymerases. II. High-density labeling of natural DNA.

Authors:  Taurai Tasara; Bernhard Angerer; Martine Damond; Holger Winter; Sabine Dörhöfer; Ulrich Hübscher; Mario Amacker
Journal:  Nucleic Acids Res       Date:  2003-05-15       Impact factor: 16.971

5.  Simultaneous, Single-Cell Measurement of Messenger RNA, Cell Surface Proteins, and Intracellular Proteins.

Authors:  Kah Teong Soh; Joseph D Tario; Sean Colligan; Orla Maguire; Dalin Pan; Hans Minderman; Paul K Wallace
Journal:  Curr Protoc Cytom       Date:  2016-01-06

6.  RNA Flow Cytometry Using the Branched DNA Technique.

Authors:  Kah Teong Soh; Paul K Wallace
Journal:  Methods Mol Biol       Date:  2018

7.  Cyanine dye dUTP analogs for enzymatic labeling of DNA probes.

Authors:  H Yu; J Chao; D Patek; R Mujumdar; S Mujumdar; A S Waggoner
Journal:  Nucleic Acids Res       Date:  1994-08-11       Impact factor: 16.971

8.  A long-wavelength biolabeling reagent based on the oxonol fluorophore.

Authors:  P L Southwick; K M Hahn; J Chao; P L Perry; A S Wagman; M Wagner; A S Waggoner
Journal:  J Fluoresc       Date:  1995-06       Impact factor: 2.217

9.  Assessment of different permeabilization methods of minimizing damage to the adherent cells for detection of intracellular RNA by flow cytometry.

Authors:  Zahra Amidzadeh; Abbas Behzad Behbahani; Nasrollah Erfani; Sedigheh Sharifzadeh; Reza Ranjbaran; Leili Moezi; Farzaneh Aboualizadeh; Mohammad Ali Okhovat; Parniyan Alavi; Negar Azarpira
Journal:  Avicenna J Med Biotechnol       Date:  2014-01

10.  Stabilization of cellular RNA in blood during storage at room temperature: a comparison of cell-free RNA BCT(®) with K3EDTA tubes.

Authors:  Kausik Das; Sheila E Norton; Jodi R Alt; Gary D Krzyzanowski; Thomas L Williams; M Rohan Fernando
Journal:  Mol Diagn Ther       Date:  2014-12       Impact factor: 4.074
  10 in total

北京卡尤迪生物科技股份有限公司 © 2022-2023.