[en] Circular dichroism and fluorescence polarization anisotropy are important tools for characterizing biomolecular systems. Both are used extensively in kinetic experiments involving stopped- or continuous flow systems as well as titrations and steady-state spectroscopy. This paper presents the theory for determining circular dichroism and fluorescence polarization anisotropy simultaneously, thus insuring the two parameters are recorded under exactly the same conditions and at exactly the same time in kinetic experiments. The approach to measuring circular dichroism is that used in almost all conventional dichrographs. Two arrangements for measuring fluorescence polarization anisotropy are described. One uses a single fluorescence detector and signal processing with a lock-in amplifier that is similar to the measurement of circular dichroism. The second approach uses classic ''T'' format detection optics, and thus can be used with conventional photon-counting detection electronics. Simple extensions permit the simultaneous measurement of the absorption and excitation intensity corrected fluorescence intensity

[en] The advantages of synchrotron radiation in several types of spectroscopy, microscopy and diffraction studies are clear. The availability of synchrotron radiation will expand rapidly in the early 1980's as experimental programs start at the new generation of dedicated storage rings

[en] This chapter describes the measurement of circular dichroism (CD) for absorption due to transitions between two distinct electronic states. This is distinguished from absorption of lower energy photons, which are associated with changes of only the vibrational modes of the absorber and from the absorption of higher energy photons, which may result in ionizations. From the instrumental viewpoint, the chapter describes the measurement of CD that can be recorded using a photomultiplier or avalanche photodiode to quantify the intensity of a light beam, a photoelastic modulator to periodically alter the beam's polarization, and a monochromator located between the light source and the modulator. Using either criterion, the focus is on the spectral domain spanning about a decade in wavelength (photon energy) from roughly 1.2 (micro)m (1 eV ∼ 160 zJ) in the near infrared to 120 nm (10 eV ∼ 1.6 aJ) in the vacuum ultraviolet (VUV). In the near infrared, there is overlap between the domain of electronic and purely vibrational transitions, the use of photomultipliers or avalanche photodiodes versus solid state detectors and dispersive versus Fourier-transform spectrometers. There is also some overlap in the VUV with synchrotron beamlines that use arrays of magnets called 'insertion devices' to cause the emitted synchrotron radiation to be elliptically polarized. To my knowledge, no single spectrometer spans this entire spectral domain discussed here, and the vast majority of laboratory instruments come nowhere close to either the upper or lower limit. However, similar analytical approaches and types of instrumentation are employed throughout this spectral domain and thus are logically treated together. The focus in this chapter is on the measurement of CD resulting from the inherent chirality of the absorbing system. Several spectroscopic methods that are closely related in terms of science or instrumentation are treated in other chapters. These include magnetic circular dichroism (MCD), linear dichroism (LD), optical rotary dispersion (ORD), fluorescence detected circular dichroism (FDCD), and circularly polarized luminescence (CPL). A basic CD instrument of the type described here can be configured by temporary alterations of the sample compartment, an additional or repositioned detector and modified electronics to perform many of the important experiments in the visible and UV regions. These include unpolarized absorption and total fluorescence in addition to most of the experiments mentioned above. Except for absorption, such extensions of the basic technology will not be discussed here. Other reviews of instrumentation related to CD have appeared, some containing information complementary to that included here.

[en] Gel electrophoresis is one of the most powerful and widely used methods for the separation of DNA. During the last decade, instruments have been developed that accurately quantitate in digital form the distribution of materials in a gel or on a blot prepared from a gel. In this paper, I review the various physical properties that can be used to quantitate the distribution of DNA on gels or blots and the instrumentation that has been developed to perform these tasks. The emphasis here is on DNA, but much of what is said also applies to RNA, proteins and other molecules. 36 refs

[en] This review examines the possibilities for biological research using the three ultraviolet free-electron lasers that are nearing operational status in the US. The projected operating characteristics of major interest in biological research of the free-electron lasers at Brookhaven National Laboratory, the Thomas Jefferson National Accelerator Facility, and Duke University are presented. Experimental applications in the areas of far- and vacuum ultraviolet photophysics and photochemistry, structural biology, environmental photobiology, and medical research are discussed and the prospects for advances in these areas, based upon the characteristics of the new ultraviolet free-electron lasers, are evaluated

No abstract available

[en] The SUPERB experiment at the SURF II storage ring uses synchrotron radiation as a source of uv photons to measure circular dichroism (CD) of biological molecules. Conventional CD instruments are limited by the lack of stable laboratory continuum sources capable of providing large photon fluxes shortward of 200 nm. Synchrotron radiation overcomes these limitations and enables one to extend CD measurements down to the limit of the window transmission cutoff, which is 125 nm for CaF₂ used in the present configuration. Some molecules of biophysical interest, DNA and proteins for example, exhibit large CD effects below 200 nm, which reflect specific molecular conformations. CD is a sensitivie probe of molecular structure and enables us to learn how these molecules behave under various external conditions. In the future we plan to increase the measurement capabilities of SUPERB to include magnetic circular dichroism, fluorescence spectroscopy, and fluorescence lifetime measurements

[en] A weeklong workshop focusing on circular dichroism (CD) spectroscopy using both conventional and synchrotron sources (SRCD) was presented at the National Synchrotron Light Source at Brookhaven National Laboratory from June 23-27, 2008.

[en] Measurements of circular dichroism (CD) in the UV and vacuum UV have used photoelastic modulators (PEMs) for high sensitivity (to about 10^-6). While a simple technique for wavelength calibration of the PEMs has been used with good results, several features of these calibration curves have not been understood. The authors have calibrated a calcium fluoride PEM and a lithium fluoride PEM using the National Synchrotron Light Source (NSLS) at Brookhaven National Laboratory as a light source. These experiments showed calibration graphs that are linear bit do not pass through the graph origin. A second ''multiple pass'' experiment with laser light of a single wavelength, performed on the calcium fluoride PEM, demonstrates the linearity of the PEM electronics. This implies that the calibration behavior results from intrinsic physical properties of the PEM optical element material. An algorithm for generating calibration curves for calcium fluoride and lithium fluoride PEMs has been developed. The calibration curves for circular dichroism measurement for the two PEMs investigated in this study are given as examples

[en] Depletion of stratospheric ozone will increase the solar ultraviolet radiation in the range from 290-320 nm (UVB) that reaches the surface of the earth, placing an increased UV burden on exposed organisms. One consequence of increased UVB may be decreased productivity of crop plants. A principal lesion caused by UV in DNA is the cyclobutyl pyrimidine dimer. We have adapted a method for measuring these dimers in nanogram quantities of non-radioactive DNA for use in UV-irradiated plants. We find that biologically relevant doses of broad band UVB radiation induce easily detectable frequencies of pyrimidine dimers in the DNA of irradiated alfalfa sprout leaves and that the dose response for dimer formation is linear up to doses of at least 690 J/m². We also find easily measurable frequencies of dimers in the leaves of seedlings grown in glass filtered sunlight but not exposed to additional UVB, suggesting that significant number of dimers are formed in plants exposed to normal sunlight. 27 refs., 3 figs., 1 tab