Improved Underwater Excitation-Emission Matrix Fluorometer

A compact, high-resolution, two-dimensional excitation-emission matrix fluorometer (EEMF) has been designed and built specifically for use in identifying and measuring the concentrations of organic compounds, including polluting hydrocarbons, in natural underwater settings. Heretofore, most EEMFs have been designed and built for installation in laboratories, where they are used to analyze the contents of samples collected in the field and brought to the laboratories. Because the present EEMF can be operated in the field, it is better suited to measurement of spatially and temporally varying concentrations of substances of interest. In excitation-emission matrix (EEM) fluorometry, fluorescence is excited by irradiating a sample at one or more wavelengths, and the fluorescent emission from the sample is measured at multiple wavelengths. When excitation is provided at only one wavelength, the technique is termed one-dimensional (1D) EEM fluorometry because the resulting matrix of fluorescence emission data (the EEM) contains only one row or column. When excitation is provided at multiple wavelengths, the technique is termed two-dimensional (2D) EEM fluorometry because the resulting EEM contains multiple rows and columns. EEM fluorometry - especially the 2D variety - is well established as a means of simultaneously detecting numerous dissolved and particulate compounds in water. Each compound or pool of compounds has a unique spectral fluorescence signature, and each EEM is rich in information content, in that it can contain multiple fluorescence signatures. By use of deconvolution and/or other mixture-analyses techniques, it is often possible to isolate the spectral signature of compounds of interest, even when their fluorescence spectra overlap. What distinguishes the present 2D EEMF over prior laboratory-type 2D EEMFs are several improvements in packaging (including a sealed housing) and other aspects of design that render it suitable for use in natural underwater settings. In addition, the design of the present 2D EEMF incorporates improvements over the one prior commercial underwater 2D EEMF, developed in 1994 by the same company that developed the present one. Notable advanced features of the present EEMF include the following: 1) High sensitivity and spectral resolution are achieved by use of an off-the-shelf grating spectrometer equipped with a sensor in the form of a commercial astronomical- grade 256 532-pixel charge-coupled-device (CCD) array. 2) All of the power supply, timing, control, and readout circuits for the illumination source and the CCD, ancillary environmental monitoring sensors, and circuitry for controlling a shutter or filter motor are custom-designed and mounted compactly on three circuit boards below a fourth circuit board that holds the CCD (see figure). 3) The compactness of the grating spectrometer, CCD, and circuit assembly makes it possible to fit the entire instrument into a compact package that is intended to be maneuverable underwater by one person. 4) In mass production, the cost of the complete instrument would be relatively low - estimated at approximately $30,000 at 2005 prices.