A high speed optical multichannel analyzer

被引:5
作者
Cole, JW
Hendler, RW
Smith, PD
Fredrickson, HA
Pohida, TJ
Friauf, WS
机构
[1] NIH, Natl Ctr Res Resources, Bethesda, MD 20892 USA
[2] NHLBI, NIH, Bethesda, MD 20892 USA
[3] NIH, Div Comp Res & Technol, Bethesda, MD 20892 USA
来源
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 1997年 / 35卷 / 03期
关键词
optical multichannel analyzer; reaction mechanism; cytochrome oxidase; bacteriorhodopsin;
D O I
10.1016/S0165-022X(97)00037-7
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An optical multichannel analyzer capable of recording spectra at sampling rates up to 100 kHz is described. The instrument, designed to gather data on the kinetic reaction mechanisms of biological preparations such as cytochrome oxidase and bacteriorhodopsin, features a massively parallel approach in which each photosensing element of the detector array has a dedicated amplifier, integrator, analog to digital converter, and sample buffer. The design has 92 such elements divided in two separate arrays, each of which sits at the focal plane of a 1/4 m Ebert spectrometer. The spectrometers may be tuned to cover independent, 130 nm wide, regions of the spectrum from 350 nm to 900 nm with a dispersion of 2.8 nm per clement. Each detection channel has 12-bit resolution with an electronic dark count of 1 count and may be sampled 1024 times during a single experiment with dynamically variable sampling intervals from 10 mu s to several seconds. Time averaging of up to thousands of consecutive laser-initiated kinetic cycles allows analyses of spectral changes <0.001 optical density units. A personal computer with custom software provides a number of features: entry of experiment parameters; transfer of data from temporary buffers to permanent files; real time display; multiple spectrum averaging; and control and synchronization of associated system hardware. Optical fibers or lenses provide coupling from a parabolic reflector Xenon are monitoring light source, through the sample chamber, to the entry slit of the monochromator. The instrument has been used for extensive studies on the rapid kinetics and definition of reaction sequences of the energy transducing enzymes cytochrome oxidase and bacteriorhodopsin. Some results from these studies are discussed. (C) 1997 Elsevier Science B.V.
引用
收藏
页码:161 / 174
页数:14
相关论文
共 6 条
[1]   Multichannel analysis of single-turnover kinetics of cytochrome aa(3) reduction of O-2 [J].
Bose, S ;
Hendler, RW ;
Shrager, RI ;
Chan, SI ;
Smith, PD .
BIOCHEMISTRY, 1997, 36 (09) :2439-2449
[2]  
Hendler R. W., 1984, J BIOCHEM BIOPH METH, V28, P1
[3]   INFLUENCE OF EXCITATION-ENERGY ON THE BACTERIORHODOPSIN PHOTOCYCLE [J].
HENDLER, RW ;
DANCSHAZY, Z ;
BOSE, S ;
SHRAGER, RI ;
TOKAJI, Z .
BIOCHEMISTRY, 1994, 33 (15) :4604-4610
[4]   MULTIWAVELENGTH ANALYSIS OF THE KINETICS OF REDUCTION OF CYTOCHROME-AA(3) BY CYTOCHROME-C [J].
HENDLER, RW ;
BOSE, SK ;
SHRAGER, RI .
BIOPHYSICAL JOURNAL, 1993, 65 (03) :1307-1317
[5]   Control of the integral membrane proton pump, bacteriorhodopsin, by purple membrane lipids of Halobacterium halobium [J].
Mukhopadhyay, AK ;
Dracheva, S ;
Bose, S ;
Hendler, RW .
BIOCHEMISTRY, 1996, 35 (28) :9245-9252
[6]   TITRATION OF INDIVIDUAL COMPONENTS IN A MIXTURE WITH RESOLUTION OF DIFFERENCE SPECTRA, PKS, AND REDOX TRANSITIONS [J].
SHRAGER, RI ;
HENDLER, RW .
ANALYTICAL CHEMISTRY, 1982, 54 (07) :1147-1152