A 5-milliwatt He-Cd laser (IK4101R-F, Kimmon Koha) was used for Raman measurement. The laser was focused on the sample in the spinning cell (1500 rpm) from below, and Raman scattering at 90° was directed to a Raman spectrometer (Chromex, 500IS); data were collected using a liquid N2-cooled CCD detector (Roper Scientific, Spec-10:400B/LN). Indene and carbon tetrachloride were used as standards for frequency calibration. The resonance Raman spectra of supercomplex, mixtures of enzyme complexes, individual enzyme complexes, and sucrose solution were measured. For each sample 20 continuous 1-min measurements were carried out. After confirmation that there was no spectral change during 20 min of laser irradiation; all 20 spectra were combined into one spectrum. The contribution of sucrose to the Raman spectrum was subtracted with a reasonable coefficient. The absorption spectrum of each sample was monitored before and after Raman measurement to confirm the redox state of the sample.
Spec 10 400b ln
Manufactured by Teledyne
Sourced in United States
The Spec-10:400B/LN is a liquid nitrogen-cooled, back-illuminated, high-performance CCD detector designed for spectroscopic applications. It features a 1340 x 400 pixel array with 20 x 20 μm pixels, providing high quantum efficiency, low noise, and high dynamic range performance.
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Lab products found in correlation
6 protocols using spec 10 400b ln
1
Resonance Raman Spectroscopy of Enzyme Complexes
2
Resonance Raman Analysis of Molecular Vibrations
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Resonance Raman analysis was carried out by 441.6 nm laser excitation (He-Cd laser (Kimmon Koha, IK4101R-F, Tokyo, Japan). The scattered light was dispersed with a single polychromator (Chromex, 500IS, Albuquerque, NM, USA), which was detected by a liquid-nitrogen-cooled CCD detector (Roper Scientific, Spec10:400B/LN, Trenton, NJ, USA). Indene was used to calibrate Raman shifts, giving an accuracy of ±1 cm−1 for intense Raman bands.
3
Resonance Raman Spectra of Cyt c
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Resonance Raman spectra were recorded using a single monochrometer (SPEX500M, Jobin Yvon, Edison, NJ) equipped with a liquid nitrogen-cooled CCD detector (Spec-10:400B/LN, Roper Scientific, Princeton, NJ). Samples were excited at a wavelength of 441.6 nm delivered by a helium-cadmium laser (IK5651R, Kimmon Koha, Tokyo, Japan).
The laser power at the sample point was adjusted to 5 mW. Raman shifts were calibrated using indene and CCl4. The accuracy of the well-defined Raman band peak positions was ± 1
The PEG 4000 concentration was varied between 0 and 5%, and the Cyt c concentration was 50 M.
The laser power at the sample point was adjusted to 5 mW. Raman shifts were calibrated using indene and CCl4. The accuracy of the well-defined Raman band peak positions was ± 1
The PEG 4000 concentration was varied between 0 and 5%, and the Cyt c concentration was 50 M.
4
Raman and EPR Spectroscopic Analysis
5
Ultrafast Spectroscopy with High Temporal Resolution
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The fs-TA spectrometer with a 120 fs time resolution, as determined as the full width at half maximum (FWHM) of instrumental response function (IRF), was described in detail elsewhere.74 (link),75 (link) Briefly, a regenerative amplifier (SPTF-100F-1KHPR, Spectra Physics) combined with an optical parametric amplifier (OPA) provided the actinic laser at a desired wavelength with a spectral bandwidth of 10 nm (FWHM). A white-light continuum probe was generated from a sapphire plate of 3 mm thickness and, after interrogating the excited sample, was detected with a liquid-nitrogen cooled charge-coupled device (CCD; Spec-10:400B/LN, Princeton Instruments) attached to an imaging spectrograph (SpectraPro 2300i; Princeton Instruments). To minimize the possible nonlinear effects, the lowest possible excitation energy (<100 nJ/Pulse) as limited by a reasonable signal-to-noise ratio was applied to the samples. The laser source was run at a repetition rate of 100 Hz to ensure that each pulse excited the dynamically relaxed sample. The sample OD at an excitation wavelength was adjusted to 0.3–0.5 in terms of an optical path length of 1 mm. Steady-state absorption spectra were checked for the samples before and after the measurements, and no appreciable sample degradation was observed. All the measurements were carried out at room temperature (296 K).
6
Femtosecond Fluorescence Experiments
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The femtosecond fluorescence experiments were conducted on a Kerr shutter set-up 36, 37 based on an oscillator-amplifier system (Tsunami-Spitfire Pro F, Spectra Physics, 1 kHz, 800 nm, 100 fs). The 400 nm excitation pulses for the Z-form were generated from the laser fundamental via second harmonic generation. The Kerr gate pulses (1300 nm) were generated using a home-built two stage OPA. 38 The fluorescence was detected using a Spectrograph (Acton Research, SpektraPro 2358) and a CCD camera (Princeton Instruments, Spec-10:400B/LN). The wavelengthdependent time zero dispersion was corrected by using the Sellmeier equation. 39, 40
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