Multichannel Spectrophotometer

The present invention relates to spectrophotometers and more particularly to spectrophotometers for blood chemistry analysis.

In the field of spectrophotometers, it has been the general practice to employ a source of radiant energy along with spectral selective elements to produce a single channel for measurement of spectral responses of samples to be measured. Multichannel spectrophotometers have been assembled using a common source of radiant energy with separate spectral selective elements to determine the spectral sensitivity of each channel. If a multiplicity of channels are required such as in blood chemistry analysis, the less common elements per channel, the more expensive and costly each channel becomes. Also, it becomes necessary to wait for a particular channel to become available to use its region of spectral interest for other measurements. Since the greatest application for multichannel spectrophotometers is in blood chemistry analysis, waiting can cause considerable delay in completing all the tests required on a given blood sample. In this general field of application, the spectrophotometers define the spectrum of interest by a single narrow band filter usually associated with the input to the radiant energy detector. Although such devices have served the purpose, they have not proved entirely satisfactory under all conditions of service for the reasons outlined above and furthermore, unless complete duplicate filter systems are provided only one at a time measurements at a given wave length can be measured or there is filter sharing or switching. These problems are overcome by the present invention.

In another analyzer, the system whirls the samples, contained in thirty cuvettes arranged around the periphery of a rotating disk. As the rotor picks up speed the centrifugal force transfers reagents from an inner disc to the cuvettes, which have transparent tops and bottoms. The chemical reactions are measured "on the fly" as the cuvettes containing the sera being measured pass through a beam from a common source lamp mounted at the top of the apparatus. The beam is spectrally filtered for the proper measurement at each detector. The rotor is timed so that the exposure of each chemical reaction in the light beam is long enough that its absorbance peak can be measured, yet is short enough to track the thirty kinetic reactions in real time.

Those concerned with the development of clinical blood analyzers have long recognized the need for a multichannel spectrophotometer which can be simultaneously utilized to measure the results at a multiplicity of sample stations. The present invention fulfills this need.

One of the most critical problems concerning designers of multichannel spectrophotometers has been the ability to easily and simply limit and define the spectral areas of sensitivity of each channel and to eliminate the problems of stray interfering radiant energy. The present invention fulfills this need.

The general purpose of this invention is to provide a multichannel spectrophotometer which embraces all the advantages of similarly employed spectrophotometer devices and possesses none of the aforedescribed disadvantages. To attain this the present invention contemplates a unique dual rotating filter wheel assembly, one on the source and one on the detector sides of the sample, whereby the effects of stray light and poor spectral selectivity are avoided.

An object of the present invention is the provision of a spectrophotometer containing a multiplicity of channels which can make bichromatic measurements substantially at the same time.

Another object is to provide a multichannel spectrophotometer which shares identical source and detector filter wheels which are aligned and rotate in synchronism.

Still another object of the present invention is to provide a multichannel spectrophotometer which makes measurements at a multiplicity of test stations simultaneously.

Still another object of the present invention is to provide a multichannel spectrophotometer which spectrally filters the radiation both before and after the sample cuvette.

Other objects and many of the attendant advantages of this invention will be readily appreciated as the same become better understood by reference to the following detailed description when considered in connection with the accompanying drawings in which like reference numerals designate like parts throughout the figures thereof and wherein:

FIG. 1 illustrates an exploded end perspective view of a preferred embodiment of the invention;

FIG. 2 shows the optical path of one of the channels of the multichannel spectrophotometer illustrated in FIG. 1;

FIG. 3 illustrates a block diagram of the timing, control and display electronics associated with each channel of the spectrophotometer;

FIG. 4 shows the automatic zero control circuitry of FIG. 3;

FIG. 5 illustrates the circuit details of the automatic gain control circuitry of FIG. 3; and

FIG. 6 illustrates the typical pulse train received through the filter wheel system at the photodetector output.

Referring now to the drawings, wherein like reference characters designate like or corresponding parts throughout the several views, there is shown in FIG. 1 a lamp 11 centered in a parabolic reflector 13. The electromagnetic radiant energy from lamp 11 is reflected from parabolic reflector 13 through heat baffles 15 to a lens disk 17. Heat baffles 15 have a circular pattern of circular holes cut near the periphery thereof to form a multiplicity of radiant energy beams from the energy reflected by parabolic reflector 13 from lamp 11. Lens disk 17 contains a series of lenses and mountings 17a-17h mounted around the outer edge thereof and aligned with the circular holes of heat baffles 15. These lens assemblies collect and direct the electromagnetic energy from lamp 11 and heat baffles 15 through rotating filter wheel 19 into electromagnetic energy conduits or light pipes 23a through 23h. An optical encoder 21 identifies the filter wheel positions so that the different filters arranged in a circular pattern along the periphery of the filter wheel are identified and known by the position of the wheel. Each lens of the lens assembly 17a-h focuses its electromagnetic energy beam on a respective light pipe 23a- h where each light pipe then transmits its corresponding selected spectrum of energy to a cuvette 25a-h for transmission therethrough. The spectral radiant energy outputs of each of the cuvettes is transmitted into a respective detector electromagnetic energy conduit 27a-h where these detector light pipes or electromagnetic energy conduits 27a-h direct their energy, respectively, through a detector filter wheel 29 which is identical with source filter wheel 19. Filter wheel 19 is connected by a shaft 34 to a synchronous motor 36 which also is connected by shaft 38 to filter wheel 29. The output of each filter segment of detector filter wheel 29 is directed to photodetectors 31a-h. Each photodetector is connected to electronic section 33 where the signals are detected and processed for display. Since source filter wheel 19 and detector filter wheel 29 are identical as far as filter segments are concerned and rotate in aligned synchronism, optical encoder then informs electronics 33 of the exact position of each filter on each wheel at any point in time.

It should be noted that one of the positions in the filter wheel is a blank and has zero transmission. This segment is used to establish the zero reference for the automatic zero circuitry discussed hereinbelow.

It should be further noted that lamp 11 may be a quartz halogen lamp made by Ushio Inc. such as JA 12V 55 WD, and electromagnetic energy conduits 23, 25 and 27 may be quartz fibers or other suitable optic material, or may be a light pipe or tube containing a transmission material of suitable optical index to contain the radiant energy therewithin.

The filters utilized on the filter wheels are very narrow band pass interference filters having band widths of 8 nm. The nano-meter wave lengths chosen for the particular filter spectral bands for blood measurement application are as follows: 340 nm, 405 nm, 450 nm, 525 nm, 550 nm, 575 nm, and 630 nm.

Although identical source and detector filter wheels are described above having the same filters in the same corresponding positions, it is contemplated within this invention that the detector filter segments can be different from the source filter segments for the purposes of certain measurements involving fluorescence where the sample cuvette is excited by electromagnetic radiation at one wavelength and the fluorescent response is measured at another. Therefore the description of FIG. 1 is to include this application where the filter wheels are rotated in synchronism but are not identical in wavelength.

In FIG. 2, there is illustrated the optical path of the electromagnetic energy of one of the beams of the preferred embodiment illustrated in FIG. 1. Electromagnetic energy emitted from lamp 11 is reflected from parabolic reflector 13 through heat baffles 15 and into a plano convex lens 17a where the energy is focused through filter 19a into source electromagnetic energy conduit 23a or light pipe 23a. The electromagnetic energy transmitted by light pipe 23a is transmitted through cuvette 25a containing a sample to be analyzed and then into detector electromagnetic energy conduit or light pipe 27a and through a neutral filter 19a and band-pass filter 29a which is identical with filter 19a, except for fluorescence measurement, and finally into photo-detector 31a. It should be noted that photo-detector 31 may be of the photo multiplier type, which has a high sensitivity and a large dynamic range such as Hamamatsu photo-multiplier tube number R6487647. Since the narrow-band filters are sensitive to temperature, air is blown through heat baffles 15 (not illustrated) to control the temperature of filter wheel 19 In addition, the photo-multiplier tubes are also temperature sensitive wih respect to their spetral response and it is therefore desirable to control the temperature thereof by appropriate means. FIG. 3 illustrates a block diagram of the electronics associated with each channel of the multichannel spectrophotometer embodiment of FIG. 1. Photomultiplier 31 is connected by line 32 to a sample averaging block 35, an automatic zero block 37 and an automatic gain control block 39. Each of these blocks are under the control of timing and control registers 51a and a computer 51. Sample averaging block 35 may be a simple integrating circuit timed to receive the signal pulses from the desired channel to which photomultiplier 31 is connected and to integrate these pulses to improve the signal to noise ratio. One of the integrated signals, Io, which is the reference signal produced by the signal pulse at the desired reference wave length and the other integrated signal is I which is the sample signal produced by the pulses at the sample wave length. These are selected gates operated by the appropriate timing and control registers as the filter wheels rotate in the multichannel spectrophotometer.

The automatic zero control for a given channel is operated every time the non-transmitting or blank segment of the filter wheel is opposite photomultiplier 31 of the given channel. It establishes a reference voltage which is used to cancel the voltage measured during the filter blank thereby establishing a "true" zero when there is no input signal.

Automatic gain control 39 adjusts the gain of each photomultiplier depending upon the amplitude of the first few signal pulses received in that channel. These first few pulses are received and integrated and a voltage established which is connected to a high voltage control 41 to adjust a high voltage power supply 33 to raise or lower the high voltage on the photomultiplier to change its gain. Since the dynamic range of the pulses may vary as much as 1000 to 1, it is necessary to adjust the gain of the photo multiplier to accommodate this dynamic range.

The reference and sample output signals I and Io are fed into a multiplexer 45 along with the similar sample and reference signals from other channels to be alternately switched into a log ratio circuit 47. This circuit is described in detail in U.S. Pat. No. 3,664,744 entitled "Log Ratio Circuit For Double Beam Spectrophotometers" granted to this inventor May 23, 1972. The output of log ratio circuit 47 which now provides a signal related to the concentration of the unknown being measured, is connected to a display 49 which may be a hard copy machine or one of a variety of alph-numeric displays well known to the electronic designer.

It should be noted that computer 51 may be part of an overall computer system along with other pieces of measurement and control equipment of which the multichannel spectrophotometer is a part. Programs in the computer can be utilized to make the multichannel spectrophotometer a useful integral part of an overall measurement system or large clinical analyzer. The sync information from the source filter wheel enables the computer to know at all times the position of a given filter and the identification signal pulses appearing in any given channel.

In FIG. 4, there is illustrated the circuit details of automatic zero control 37 consisting of an operational amplifier 53 whose output is connected by a gate 55 to an integrating amplifier 57 consisting of an operational amplifier with an integrating capacitor 59. When a blank in each filter wheel comes in position for a particular channel, gate 55 is opened to transmit whatever voltage offset appears at the output of amplifier 53 for that channel and it is stored on capacitor 59 and held as an offset voltage which is applied back to the input of operational amplifier 53 to offset any voltage when the blank appears in the filter wheel for that channel.

FIG. 5 illustrates the circuit details of automatic gain control 39 of FIG. 3. A capacitor 63 is connected through gate 61 through a resistor network to line 32. Capacitor 63 is also connected through a gate 65 to a nominal bias. Capacitor 63 is further connected to the input to operational amplifier 67 whose output is connected to optical coupler 69 which optically couples a control signal to high voltage control 41. As gate 61 is operated to connect the first few pulses from the photomultiplier detector, voltage across capacitor 63 is changed changing the output voltage of amplifier 67 which in turn is coupled by optical coupler to high voltage control 41. Therefore, if the pulses are extremely large, the gain on the photomultiplier tube for that channel can be reduced to accommodate the large pulses and prevent overload of the analog circuits. This allows the photomultiplier detector to accommodate the extreme dynamic range experienced by the different spectrophotometer channels. Gate 65 enables the gain to be quickly returned to normal range from an extreme by gating a normal voltage to capacitor 63.

FIG. 6 illustrates the sequence of pulses which may be generated at the photomultiplier output of each spectrophotometer channel. For example, the averaging circuit 35 of FIG. 3 may average 100 pulses of wave length lambda.sub.2 or the second pulse of FIG. 6 to obtain the signal value which will be used for that channel. Similarly, the same averaging is utilized to obtain the reference signal.

Operation of the preferred embodiment illustrated in FIG. 1 may be described first by reference to FIGS. 1 and 2. Lamp 11 with parabolic reflector 13 and heat baffles 15 along with lens disk 17 and lens assemblies 17a-h, form light substantially parallel beams of wide band electromagnetic radiant energy. Heat baffles 15 are required since the interference filters used in rotating filter wheel 19 are sensitive to temperature, each segment of filter wheel 19 containing an interference filter having a narrow spectral band centered at a given wave length as hereinbefore set forth. Therefore, except for the blank segment of the filter wheel, each "light pipe" 23a-h transmits a narrow band of electromagnetic energy centered at the wave length of the filter oppositely disposed that particular light pipe at that moment. The filter wheel makes 30 revolutions per second and therefore every second each light pipe 23a-23h sees repeatedly at 30.times.per second a sequential series of seven narrow bands of spectral energy and a blank reference. Light pipes 23a-h transmit this successive series of narrow bands of spectral energy to eight different test stations containing a sample cuvettes 25a-h to be analyzed. If the cuvettes contain blood serum to be analyzed, the various filters for the filter wheel will be chosen to perform the various tests required. For each test one wave length will be chosen as a reference and another as the sample wave length or bichromatic. Once these tests and wave lengths are established, then the filters can be selected for the filter wheel.

Cuvettes a-h may be part of an overall system of blood analysis and can be part of a continuous chain of cuvettes which are moved in a controlled environment from one test station to another.

The energy from cuvettes 25a-h, respectively, is directed into detector light pipes 27a-h. To prevent stray radiant energy from interfering with the desired signals and to increase the spectral selectivity of the system the detector light pipes direct their energy through a second rotating filter wheel 29 having the same identical filters as filter wheel 19 and the same identical positions. Consequently when light pipe 23a is receiving output from filter lambda one, then detector 31a is receiving energy from filter lambda one directed thereto by detector light pipe 27a. To balance the energy transmission in each of the transmission paths, FIG. 2 illustrates the use of a neutral filter 29a. Neutral filter 29a has no spectral characteristics and attenuates the energy transmitted therethrough. It merely balances or attempts to balance the flat spectral energy transmission from path to path.

It should be noted although quartz fiber optics is desired for light pipes 23 and 27, other forms of electromagnetic energy transmission conduits may be utilized. Conduits containing various liquids have proved successful and may be preferred over fiber optics under certain circumstances.

Therefore, the output of each photomultiplier detector 31 appears much as the train of pulses illustrated in FIG. 6 where the sequence repeats and repeats as the filter wheel rotates and completes each resolution.

Turning now to FIG. 3, since the filter wheels rotate at 1800 revolutions per minute, in 3 and 1/3 pulse samples are averaged by sample averaging 35 to produce an Io reference and an I sample signal from each channel. There being a sample averaging circuit 35 for each photomultiplier detector 31a-h, the concentration of the unknowns being measured for each of the channels is computed by the log ration circuit 47 shared by all the channels through multiplexer 45 and displayed by display 49.

The automatic gain control and zero control which have been previously discussed make the multichannel spectrophotometer ideally suited to blood chemistry measurements.

Although not illustrated, it should be emphasized that the temperature of the filter wheels must be controlled in order to preserve the spectral characteristics of the interference filters. Typical fans and blowers under reasonable temperature control are sufficient for this task.

It should be quite evident that the multichannel spectrophotometer can be placed under computer control and to have the desired sequence of measurements to be performed at the remote cuvette stations merely by programming the computer to operate the timing and control registers in a manner which gate the appropriate pulses into the proper electronic sections at the proper time. Therefore the multichannel spectrophotometer is completely adaptable to automatic control including test sequences, tests to be performed at various locations and data handling, processing and display.

It now should be apparent that the present invention provides a mechanical and circuit arrangement which may be employed in conjunction with a multichannel spectrophotometer for providing a highly spectrally sensitive instrument to perform a variety of tests at remote test stations.

Although particular components, etc. have been discussed in connection with a specific embodiment of a spectrophotometer constructed in accordance with the teachings of the present invention, others may be utilized. Furthermore, it will be understood that although an exemplary embodiment of the present invention has been disclosed and discussed, other applications of circuit arrangements are possible and that the embodiments disclosed may be subjected to various changes, modifications and substitutions without necessarily departing from the spirit of the invention.