Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Breaking the 200 nm limit for routine flow linear dichroism measurements using UV synchrotron radiation

Dicko, Cedric LU orcid ; Hicks, Matthew R ; Dafforn, Timothy R ; Vollrath, Fritz ; Rodger, Alison and Hoffmann, Søren V (2008) In Biophysical Journal 95(12). p.5974-5977
Abstract

The first synchrotron radiation flow linear dichroism spectra are reported. High-quality spectral data can be collected from 450 nm down to 180 nm in contrast to the practical cutoff of approximately 200 nm on benchtop instruments. State-of-the-art microvolume capillary Couette flow linear dichroism was successfully ported to a synchrotron radiation source. The sample volume required is < 50 microL. A characterization of the synchrotron radiation linear dichroism with known DNA and DNA-ligand systems is presented and the viability of the setup confirmed. Typically, wavelengths down to 180 nm are now routinely accessible with a high signal/noise ratio with little limitation from the sample concentration. The 180 nm cutoff is due to... (More)

The first synchrotron radiation flow linear dichroism spectra are reported. High-quality spectral data can be collected from 450 nm down to 180 nm in contrast to the practical cutoff of approximately 200 nm on benchtop instruments. State-of-the-art microvolume capillary Couette flow linear dichroism was successfully ported to a synchrotron radiation source. The sample volume required is < 50 microL. A characterization of the synchrotron radiation linear dichroism with known DNA and DNA-ligand systems is presented and the viability of the setup confirmed. Typically, wavelengths down to 180 nm are now routinely accessible with a high signal/noise ratio with little limitation from the sample concentration. The 180 nm cutoff is due to the quartz of the Couette cell rather than the beamline itself. We show the application of the simultaneous determination of the sample absorption spectrum to calculate the reduced linear dichroism signal. Spectra for calf thymus DNA, DNA/ethidium bromide, and DNA/4',6-diamidino-2-phenylindole systems illustrate the quality of data that can be obtained.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
keywords
Animals, Cattle, DNA/chemistry, Ligands, Spectrum Analysis/methods, Synchrotrons, Ultraviolet Rays
in
Biophysical Journal
volume
95
issue
12
pages
5974 - 5977
publisher
Cell Press
external identifiers
  • pmid:18805928
  • scopus:58749104390
ISSN
1542-0086
DOI
10.1529/biophysj.108.139964
language
English
LU publication?
no
id
02675726-7eda-4d4c-aaec-79173abc9611
date added to LUP
2019-06-28 00:28:18
date last changed
2024-04-30 15:48:42
@article{02675726-7eda-4d4c-aaec-79173abc9611,
  abstract     = {{<p>The first synchrotron radiation flow linear dichroism spectra are reported. High-quality spectral data can be collected from 450 nm down to 180 nm in contrast to the practical cutoff of approximately 200 nm on benchtop instruments. State-of-the-art microvolume capillary Couette flow linear dichroism was successfully ported to a synchrotron radiation source. The sample volume required is &lt; 50 microL. A characterization of the synchrotron radiation linear dichroism with known DNA and DNA-ligand systems is presented and the viability of the setup confirmed. Typically, wavelengths down to 180 nm are now routinely accessible with a high signal/noise ratio with little limitation from the sample concentration. The 180 nm cutoff is due to the quartz of the Couette cell rather than the beamline itself. We show the application of the simultaneous determination of the sample absorption spectrum to calculate the reduced linear dichroism signal. Spectra for calf thymus DNA, DNA/ethidium bromide, and DNA/4',6-diamidino-2-phenylindole systems illustrate the quality of data that can be obtained.</p>}},
  author       = {{Dicko, Cedric and Hicks, Matthew R and Dafforn, Timothy R and Vollrath, Fritz and Rodger, Alison and Hoffmann, Søren V}},
  issn         = {{1542-0086}},
  keywords     = {{Animals; Cattle; DNA/chemistry; Ligands; Spectrum Analysis/methods; Synchrotrons; Ultraviolet Rays}},
  language     = {{eng}},
  month        = {{12}},
  number       = {{12}},
  pages        = {{5974--5977}},
  publisher    = {{Cell Press}},
  series       = {{Biophysical Journal}},
  title        = {{Breaking the 200 nm limit for routine flow linear dichroism measurements using UV synchrotron radiation}},
  url          = {{http://dx.doi.org/10.1529/biophysj.108.139964}},
  doi          = {{10.1529/biophysj.108.139964}},
  volume       = {{95}},
  year         = {{2008}},
}