Methods and apparatus for vacuum ultraviolet (VUV) or shorter wavelength circular dichroism spectroscopy
A highly efficient vacuum ultraviolet circular dichroism spectrometer is provided; the spectrometer suitable for laboratory use or for integration into a beam line at a synchrotron radiation facility. In one embodiment, a spectroscopic circular dichroism instrument is provided; the instrument configured so as to enable circular dichroism data to be simultaneously obtained for multiple wavelengths of light. The instrument may be further configured to operate in at least a portion of the vacuum ultraviolet wavelength region.
1. A spectroscopic circular dichroism instrument, comprising:
a light source which generates a multi-wavelength light beam comprised of light of a plurality of wavelengths;
a region of the spectroscopic circular dichroism instrument for the placement of a sample from which a circular dichroism measurement is to be obtained by exposure of a sample to the multi-wavelength light beam;
a constant phase difference compensator optically coupled to the multi-wavelength light beam, the constant phase difference compensator introducing a phase difference between two orthogonal polarization components of the multi-wavelength light beam;
an optical element coupled to the multi-wavelength light beam, the optical element selecting a linearly polarized component of the multi-wavelength light beam; and
a detector coupled to the multi-wavelength light beam to provide a circular dichroism measurement of the sample, the instrument being capable of providing the circular dichroism measurement simultaneously for multiple wavelengths,
wherein the compensator is rotatable,
wherein circular dichroism spectra and linear dichroism spectra are extracted from a detected intensity signal, and
wherein circular dichroism spectra are extracted from dc and 2ω components of the detected intensity signal and linear dichroism spectra are extracted from the dc and 4ω components of the detected intensity signal, wherein ω is an angular frequency of the compensator rotation.
2. The spectroscopic circular dichroism instrument of claim 1 , wherein an optical path of the instrument is configured to allow the multi-wavelength light beam to pass through the constant phase difference compensator and the optical element before exposure of the sample to the multi-wavelength light beam.
3. The spectroscopic circular dichroism instrument of claim 1 , where the constant phase difference compensator provides a circular polarization component to the multi-wavelength light beam.
4. The spectroscopic circular dichroism instrument of claim 1 , wherein the sample is a protein solution.
5. The spectroscopic circular dichroism instrument of claim 4 , wherein the spectroscopic circular dichroism instrument is configured to measure the circular dichroism of spectra of the protein solution, the spectroscopic circular dichroism instrument further configured to analyze the circular dichroism spectra to obtain a protein secondary structure.
6. The spectroscopic circular dichroism instrument of claim 1 , further comprising a depolarizer coupled to the multi-wavelength light beam.
7. The spectroscopic circular dichroism instrument of claim 1 , further comprising a second compensator and second optical element, the second optical element selecting a linearly polarized component of the multi-wavelength light beam.
8. A spectroscopic circular dichroism instrument, comprising:
a light source which generates a multi-wavelength light beam comprised of light of a plurality of wavelengths;
a region of the spectroscopic circular dichroism instrument for the placement of a sample from which a circular dichroism measurement is to be obtained by exposure of a sample to the multi-wavelength light beam;
a compensator optically coupled to the multi-wavelength light beam, the compensator introducing a phase difference between two orthogonal polarization components of the multi-wavelength light beam;
an optical element coupled to the multi-wavelength light beam, the optical element selecting a linearly polarized component of the multi-wavelength light beam; and
a detector coupled to the multi-wavelength light beam to provide a circular dichroism measurement of the sample, the instrument being capable of providing the circular dichroism measurement simultaneously for multiple wavelengths,
wherein an optical path of the instrument is configured to allow the multi-wavelength light beam to pass through the compensator and the optical element after exposure of the sample to the multi-wavelength light beam,
wherein the compensator is rotatable,
wherein circular dichroism spectra and linear dichroism spectra are extracted from a detected intensity signal, and
wherein circular dichroism spectra are extracted from dc and 2ω components of the detected intensity signal and linear dichroism spectra are extracted from the dc and 4ω components of the detected intensity signal, wherein ω is an angular frequency of the compensator rotation.
9. A spectroscopic circular dichroism instrument, comprising:
a light source which generates a multi-wavelength light beam comprised of light of a plurality of wavelengths;
a region of the spectroscopic circular dichroism instrument for the placement of a sample from which a circular dichroism measurement is to be obtained by exposure of a sample to the multi-wavelength light beam;
a compensator optically coupled to the multi-wavelength light beam, the compensator introducing a phase difference between two orthogonal polarization components of the multi-wavelength light beam;
an optical element coupled to the multi-wavelength light beam, the optical element selecting a linearly polarized component of the multi-wavelength light beam; and
a detector coupled to the multi-wavelength light beam to provide a circular dichroism measurement of the sample, the instrument being capable of providing the circular dichroism measurement simultaneously for multiple wavelengths,
wherein the compensator is rotatable and provides a circular polarization component to the multi-wavelength light beam,
wherein circular dichroism spectra and linear dichroism spectra are extracted from a detected intensity signal, and
wherein circular dichroism spectra are extracted from dc and 2ω components of the detected intensity signal and linear dichroism spectra are extracted from the dc and 4ω components of the detected intensity signal, wherein ω is an angular frequency of the compensator rotation.
10. A spectroscopic circular dichroism instrument, comprising:
a light source which generates a multi-wavelength light beam comprised of light of a plurality of wavelengths;
a region of the spectroscopic circular dichroism instrument for the placement of a sample from which a circular dichroism measurement is to be obtained by exposure of a sample to the multi-wavelength light beam;
a compensator optically coupled to the multi-wavelength light beam, the compensator introducing a phase difference between two orthogonal polarization components of the multi-wavelength light beam;
an optical element coupled to the multi-wavelength light beam, the optical element selecting a linearly polarized component of the multi-wavelength light beam; and
a detector coupled to the multi-wavelength light beam to provide a circular dichroism measurement of the sample, the instrument being capable of providing the circular dichroism measurement simultaneously for multiple wavelengths,
wherein the compensator is rotatable,
wherein circular dichroism spectra and linear dichroism spectra are extracted from a rotation dependent intensity signal, and
wherein circular dichroism spectra are extracted from dc and 2ω components of the rotation dependent intensity signal and linear dichroism spectra are extracted from the dc and 4ω components of the rotation dependent signal, wherein ω is an angular frequency of the compensator rotation.
11. A method for performing a dichroism measurement, the dichroism measurement being a circular dichroism measurement, a linear dichroism measurement or both a circular and linear dichroism measurement, comprising:
generating a light beam comprised of multiple wavelengths;
exposing a measurement sample with said light beam and passing the light beam through the sample;
inducing a phase difference between orthogonal polarization components of an emerging light beam emerging from the sample;
selecting a polarization component of the emerging light beam;
detecting an intensity of a polarization component output light beam at multiple wavelengths; and
extracting a dichroism measurement from the intensity of the polarization component output light beam for each of the multiple wavelengths,
wherein the phase difference between the orthogonal polarization components is induced by use of an optical compensator,
wherein the optical compensator is rotated during measurement, generating time- and wavelength-dependent intensity data, and
wherein circular dichroism spectra are extracted from dc and 2ω components of a detected intensity signal and linear dichroism spectra are extracted from the dc and 4ω components of the detected intensity signal, wherein ω is an angular frequency of the compensator rotation.
12. The method of claim 11 , wherein the dichroism measurement is a circular dichroism measurement.
13. The method of claim 11 , wherein the dichroism measurement is a linear dichroism measurement.
14. The method of claim 11 , wherein the dichroism measurement is both a circular dichroism measurement and a linear dichroism measurement.
15. The method of claim 11 , wherein the sample is a protein solution.
16. The method of claim 15 , further comprising analyzing a circular dichroism spectra to obtain data regarding a protein secondary structure.
17. The method of claim 11 , wherein the polarization component is selected by use of an analyzer/polarizer.