Mp. Russell et al., DESIGN AND PERFORMANCE OF AN ULTRAVIOLET RESONANCE RAMAN SPECTROMETERFOR PROTEINS AND NUCLEIC-ACIDS, Biophysical journal, 68(4), 1995, pp. 1607-1612
We describe an ultraviolet resonance Raman (UVRR) spectrometer appropr
iate for structural studies of biological macromolecules and their ass
emblies. instrument design includes the following features: a continuo
us wave, intracavity doubled, ultraviolet laser source for excitation
of the Raman spectrum; a rotating cell (or jet source) for presentatio
n of the sample to the laser beam; a Cassegrain optic with f/1.0 apert
ure for collection of the Raman scattering; a quartz prism dispersing
element for rejection of stray light and Rayleigh scattering; a 0.75-m
single grating monochromator for dispersion of the Raman scattering;
and a liquid-nitrogen-cooled, charge-coupled device for detection of t
he Raman photons. The performance of this instrument, assessed on the
basis of the observed signal-to-noise ratios, the apparent resolution
of closely spaced spectral bands, and the wide spectrometer bandpass o
f 2200 cm(-1), is believed superior to previously described UVRR spect
rometers of similar design. Performance characteristics of the instrum
ent are demonstrated in UVRR spectra obtained from standard solvents,
p-ethylphenol, which serves as a model for the tyrosine side chain, th
e DNA nucleotide deoxyguanosine-5'-monophosphate, and the human tumor
necrosis factor binding protein, which is considered representative of
soluble globular proteins.