Real-time imaging of single DNA molecules with fluorescence microscopy.

A fluorescence microscopy technique was used to image the dynamics of individual DNA molecules. Lambda, calf thymus, cosmid (circular), and T4 DNA were studied with the fluorescent dye acridine orange. Experiments with DNAase I were conducted, and the results indicate that these observations corresp...

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Main Authors: Houseal, T W, Bustamante, C, Stump, R F, Maestre, M F
Formato: Artigo
Idioma:English
Publicado: 1989
Assuntos:
Research Article
Acceso en liña:https://ncbi.nlm.nih.gov/pmc/articles/PMC1280503/
https://ncbi.nlm.nih.gov/pubmed/2790136
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spelling pubmed-12805032006-01-26 Real-time imaging of single DNA molecules with fluorescence microscopy. Houseal, T W Bustamante, C Stump, R F Maestre, M F Biophys J Research Article A fluorescence microscopy technique was used to image the dynamics of individual DNA molecules. Lambda, calf thymus, cosmid (circular), and T4 DNA were studied with the fluorescent dye acridine orange. Experiments with DNAase I were conducted, and the results indicate that these observations correspond to DNA molecules. The results of experiments with circular DNA provide strong evidence that these were single DNA molecules. Molecules were observed free in solution or attached to a glass or copper surface at one or several points. The Brownian motion of these molecules was observed, indicating that DNA in solution exists in a partially supercoiled state. Some molecules appeared stretched and were attached to the surface by their termini; the lengths of these molecules were measured. Such molecules also exhibited elastic behavior upon breaking. The power of this technique is demonstrated in images of cosmid DNA molecules, catenanes, and DNA extending from T4 phage particles. These results suggest immediate applications to molecular biology, such as examining the dynamics of protein-DNA interactions. Areas of ongoing research are discussed. 1989-09 /pmc/articles/PMC1280503/ /pubmed/2790136 Text en
institution US National Library of Medicine
collection PubMed Central
language English
format Article
topic Research Article
spellingShingle Research Article
Houseal, T W
Bustamante, C
Stump, R F
Maestre, M F
Real-time imaging of single DNA molecules with fluorescence microscopy.
description A fluorescence microscopy technique was used to image the dynamics of individual DNA molecules. Lambda, calf thymus, cosmid (circular), and T4 DNA were studied with the fluorescent dye acridine orange. Experiments with DNAase I were conducted, and the results indicate that these observations correspond to DNA molecules. The results of experiments with circular DNA provide strong evidence that these were single DNA molecules. Molecules were observed free in solution or attached to a glass or copper surface at one or several points. The Brownian motion of these molecules was observed, indicating that DNA in solution exists in a partially supercoiled state. Some molecules appeared stretched and were attached to the surface by their termini; the lengths of these molecules were measured. Such molecules also exhibited elastic behavior upon breaking. The power of this technique is demonstrated in images of cosmid DNA molecules, catenanes, and DNA extending from T4 phage particles. These results suggest immediate applications to molecular biology, such as examining the dynamics of protein-DNA interactions. Areas of ongoing research are discussed.
author Houseal, T W
Bustamante, C
Stump, R F
Maestre, M F
author_facet Houseal, T W
Bustamante, C
Stump, R F
Maestre, M F
author_sort Houseal, T W
title Real-time imaging of single DNA molecules with fluorescence microscopy.
title_short Real-time imaging of single DNA molecules with fluorescence microscopy.
title_full Real-time imaging of single DNA molecules with fluorescence microscopy.
title_fullStr Real-time imaging of single DNA molecules with fluorescence microscopy.
title_full_unstemmed Real-time imaging of single DNA molecules with fluorescence microscopy.
title_sort real-time imaging of single dna molecules with fluorescence microscopy.
publishDate 1989
url https://ncbi.nlm.nih.gov/pmc/articles/PMC1280503/
https://ncbi.nlm.nih.gov/pubmed/2790136
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