US3555284A - Multistation, single channel analytical photometer and method of use - Google Patents
Multistation, single channel analytical photometer and method of use Download PDFInfo
- Publication number
- US3555284A US3555284A US784739A US3555284DA US3555284A US 3555284 A US3555284 A US 3555284A US 784739 A US784739 A US 784739A US 3555284D A US3555284D A US 3555284DA US 3555284 A US3555284 A US 3555284A
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- United States
- Prior art keywords
- chambers
- rotor assembly
- rotor
- sample analysis
- series
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N21/07—Centrifugal type cuvettes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
- B04B—CENTRIFUGES
- B04B5/00—Other centrifuges
- B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
- B04B5/0407—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers for liquids contained in receptacles
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/11—Automated chemical analysis
- Y10T436/111666—Utilizing a centrifuge or compartmented rotor
Definitions
- Solution handling systems cgmp 'ising sets of intercon- 356/197- 336/3461 350/3331 23/252 nected, solution-accepting chambers are disposed generally in Int. radial alignment the sample hambers forming the cuvette system.
- the solution-accepting chambers of the solution Fleld ofSearch handling ystems are shaped and ized to retain when 2233.
- a single light source and a 253 photodetecting unit are aligned with the windows to determine chemical species concentrations by light absorbancy in [56] References cued the samples contained in the cuvettes.
- Means for receiving the UNITED STATES PATENTS output from the photodetecting unit are provided for in- 2,543,303 2/1951 Rubissow 264/311 dividually indicating the phototransmittance of the samples 2.552.107 5/1951 Miller et al.
- photometric should not be considered in a restrictive sense as it is intended to be generic to the terms colorimetric, fluorometric and spectrometric. Consistent with such usage, the term photometer is also used in a broad sense to include those devices sometimes referred to in the art as colorimeters, fluorometers and spectrometers.
- light as used herein includes radiant energy in both the visible and invisible spectrums as well as radiant energy restricted to specific wave lengths. Thus the invention should be understood to encompass systems which utilize different types of radiation to accomplish the measurement desired. The need for a photometric system capable of performing analyses on a large number of discrete samples simultaneously has long existed in clinical and analytical laboratories.
- Still another deficiency in many prior art photometric instruments is that volumes of samples, enzymes and other expensive reagents larger than desirable are required. This deficiency is in some cases the result of continuous flow monitoring systems which are inefficient when small numbers of samles are analyzed. A further deficiency is the undesirability of handling many small, discrete volumes of samples and reagents individually and mixing them at timed intervals.
- Another object of the invention is to provide a photometric system wherein the steps of volumetric measurement, liquid transfer, solution mixing, reaction, photometric measurement, and data reduction may be performed within a single system.
- a photometer for simultaneously determining the presence of a common substance in a multiplicity of discrete samples.
- a multiplicity of sample chambers with axially aligned transparent windows are arranged within a centrifuge rotor to provide a rotary cuvette system.
- Solution handling systems comprising sets of interconnected, solution-accepting chambers are disposed generally in radial alignment with the sample chambers forming the cuvette system.
- the solutionaccepting chambers 'of the solution handling systems are shaped and sized to retain liquid when the rotor is at rest, and to release the liquid to the cuvettes when the rotor is spinning.
- a single light source and photodetecting unit are aligned with the windows to determine chemical species concentrations by light absorbency in the samples contained in the cuvettes.
- Means receiving the output from the photodetecting unit are provided for indicating the phototransmittance of the samples within each cuvette.
- FIG. 1 schematically illustrates a photometric 7 system designed in accordance with the present invention.
- FIG. 2 is an exploded perspective view, in section of the rotor used in the system illustrated in FIG. 1. 7
- FIG. 3 is an oscillogram obtained with the system of FIG. I using a 660 ,u filter and distilled water in all cuvettes. v
- FIG. 4 is an oscillogram obtained with the system of FIG. 1 using a 660 p. filter wherein a uniform solution containing water, bovine serum albumen and bromphenol blue was introduced into cuvettes numbered 2-15 during rotation.
- FIG. 5 is an oscillogram obtained with the system of FIG. 1 using a 550 ,u. filter and a series of incremented standards in cuvettes numbered 3 1 2.
- FIG. 6 is a plot of the data obtained from FIG. 5.
- FIG. 7 is a plot of absorbencies against protein concentration obtained using the system of FIG. 1 in performing the experiment described in Example ll.
- FIG. 1 schematically illustrates an analyzer made in accordance with the invention.
- a pancake-shaped rotor assembly l illustrated in greater detail in the exploded view of FIG. 2 where like numerals are used to denote like parts, comprises a bolt flanged steel rotor body 2, glass rings 3 and 4, a slotted polytetrafluorethylene cuvette ring 5, polytetrafluorethylene retaining rings and 7, and a steel bolted flange ring 8. Rings 3, 4, 5, 6 and 7 are compressed between rotor body 2 and flange ring 8 to form a multiplicity of radially oriented cuvettes 9 in slotted cuvette ring 5.
- Spaced holes 10, axially aligned with cuvettes 9, are provided in rotor body 2, retaining rings 6 and 7, and flange ring 8 so as to pro vide axially extending passageways permitting passage of a light beam through the cuvettes.
- a centrally positioned removable transfer disc 11 is provided with small radial projections 12 which are spaced about its periphery to mate with cuvettes in ring 5.
- Handle I3 is provided to facilitate removal of transfer disc 11 from the rotor assembly.
- Transfer disc I1 is provided with a set of chambers I4 corresponding to each cuvette 9 for receiving sample liquids and reactants while the rotor is at rest.
- Chambers 14 comprise a plurality of sloping cylindrical cavities which are interconnected at their upper ends and separated by partitions I5 at their lower ends. Partitions I5 prevent mixing of the sample and reactant liquids when the rotor is at rest while permitting such liquids to pass to cuvettes 9 when the rotor is spinning.
- a passageway 16 extends from the radially outermost cavity of each chamber 14 to the periphery of a corresponding radial projection 12 to permit passage of liquid from each chamber to a corresponding cuvette when the rotor is rotated.
- a drive motor 17 sup ports rotor assembly I as well as rotating it.
- a photometric light source and projecting means is provided to project a light beam of constant intensity intersecting rotor assembly I at a point corresponding to the radial posi tions of cuvettes 9 and spaced holes 10.
- the light beam is aligned in such a manner so as to be transmitted through each hole 10 and cuvette 9 as they pass through the beam.
- the photometric light source comprises an incandescent lamp I8 with a reflecting mirror 19 disposed below the rotor assembly and oriented to reflect the light beam upward, substantially normal to the plane of rotation.
- Electronic photodetecting means 20 is disposed above rotor assembly 1 and aligned to receive light transmitted through the cuvettes during rotation. Photodetecting means 20 is designed to respond electronically with an output which is proportional to the intensity of the light transmitted from light source 18 through the cuvettes. Photodetector 20 comprises a photomultiplier tube disposed directly above the cuvette circle toreceive all light transmitted upwardly through the axially aligned openings.
- the remaining electronic components illustrated schematically in FIG. 1 include a proportional tachometer 21 which supplies a voltage signal proportional to the rotor speed to a ramp signal generator 22 which, in turn, provides a signal to a pulse scanner 23.
- a revolution detector 24 synchronizes the ramp signal frequency with the rotor speed.
- Pulse scanning means 23, synchronizable by the ramp signal generator frequency, responds proportionately to pulses originating in photodetecting means 20 and sorts the pulses therefrom as to origin.
- Pulse peak readout means 25 continuously and simultaneously indicates phototransmittance of the liquid contents reactions in the cuvettes start essentially simultaneously and may be followed continuously on an oscilloscope or other readout means 25.
- a sample and two reagents may be loaded without mixing while the rotor is at rest and then, by spinning the rotor, caused to drain centrifugally into a corresponding cuvette where they are mixed. Connections to the cuvettes are by small passageways through projections 12 as illustrated in FIGS. 1 and 2.
- the transfer disc 11 may be adapted to hold transfer tubes as described in copending application S. N. 756,265 of common assignee, or small, commercially available, disposable microliter pipettes. Such devices allowsi'ngle or multiple addition reactions to be used or reactions in whicha reaction time occurs between two additions. ln'reactions that produce precipitates, the suspended solids can'be moved .out of the optical path by centrifugal force, allowing the absorbencies of a clear supernatant to be measured.
- the radialorientation of the cuvettes causes a difference in tangential velocity to exist between the radially innermost end of each cu'vette and its radially outermost end. Rapid acceleration and deceleration of the rotor during transfer of liquid into the cuvettes cause circular flow of the liquid therein and enhance mixing. Such mixing is considered desirable as it aids the reaction between sample and reagent and provides more'uniform results.
- the rotor is accelerated rapidly to transfer fluid to the cuvettes, decelerated rapidly to facilitate mixing, and then reacfcelerated to the speed desired for testing.
- FIG. 4 is an oscillograrn showing the pattern observed using a 660 filter and distilled water in all cuvettes.
- the oscillogram was obtained from an oscilloscope which provided the pulse peak readout means 25 described in an earlier reference to FIG. 1.
- the oscillogram of FIG. 6 was obtained by providing a complete series of incremented standards in cuvettes numbered 3 through 12, with a duplication of the solution used in the cuvettes numbered 12 also being used in the cuvette numbered 14. The four remaining cuvettes contained distilled water only. Measurements were made from photographic enlargements of the patterns obsewed on the oscilloscope, and all peaks converted to l/percent T by dividing the first blank by each subsequent reading in turn. The log of HT is the absorbence which, after blank subtraction, was then multiplied by the cuvette factor to give absorbency for a one cm. path length. The data obtained in this manner from the oscillogram of FIG. 5 is plotted in FIG. 6.
- the biuret reaction for protein is a single one-reagent analysis which is of general interest and is suitable for evaluating the efficiency of the transfer discs, of mixing, and of the ability of the system to read absorbencies early in the course of the reactions.
- the Weichselbaum biuret reagent may be used with protein solutions in a range of ratios varying from 0 to 50 percent reagent in the final mixture, providing that identical solutions are used to obtain the standard curve.
- the rotor may be brought to a complete stop and the sample-reagent disc replaced. In this manner reactions depending upon sequential timed additions may be performed.
- the centrifugal capabilities of the rotor may also be employed, where desired, to sediment particulate matter or to ensure that the solutions are not turbid when their absorbencies are measured.
- rotor assemblies 1 may be fabricated with more or less cuvettes than shown or with different materials such as transparent plastics.
- the centrally positioned transfer disc may also be provided with more or less chambers for receiving sample and reactant liquids and such chambers may vary from the particular shape illustrated. It is intended rather that the invention be limited only by the scope of the appended claims.
- a photometric solution analyzer for the simultaneous determination of a common substance in a multiplicity of discrete samples comprising:
- a light source for providing a beam of light incident on said rotor assembly at a point corresponding to the radial position of said sample analysis chambers
- (1. means receiving the output from said light detecting means for continuously and simultaneously indicating the presence of said common substance within each of said sample analysis chambers.
- sample analysis chambers comprise a multiplicity of radially oriented elongated cavities disposed in a circular array about the center of rotation of said rotor assembly.
- the photometric analyzer of claim 1 wherein said means for detecting light comprises a photomultiplier tube.
- a method for photometrically analyzing a multiplicity of discrete samples to simultaneously determine the presence of a single substance therein comprising:
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Optical Measuring Cells (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US78473968A | 1968-12-18 | 1968-12-18 |
Publications (1)
Publication Number | Publication Date |
---|---|
US3555284A true US3555284A (en) | 1971-01-12 |
Family
ID=25133381
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US784739A Expired - Lifetime US3555284A (en) | 1968-12-18 | 1968-12-18 | Multistation, single channel analytical photometer and method of use |
Country Status (13)
Country | Link |
---|---|
US (1) | US3555284A (de) |
JP (1) | JPS4838274B1 (de) |
AT (1) | AT297373B (de) |
BE (1) | BE742963A (de) |
BR (1) | BR6915271D0 (de) |
CH (1) | CH502595A (de) |
DE (1) | DE1962267C3 (de) |
DK (1) | DK134795B (de) |
ES (1) | ES374546A1 (de) |
FR (1) | FR2026450A1 (de) |
GB (1) | GB1242034A (de) |
NL (1) | NL164391C (de) |
SE (1) | SE361941B (de) |
Cited By (99)
Publication number | Priority date | Publication date | Assignee | Title |
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US3656116A (en) * | 1970-05-05 | 1972-04-11 | Atomic Energy Commission | Computer interface |
US3712742A (en) * | 1969-08-05 | 1973-01-23 | Anvar | Analytical centrifuges having a photoelectric detection |
JPS4837193A (de) * | 1971-09-09 | 1973-06-01 | ||
FR2182736A1 (de) * | 1972-05-05 | 1973-12-14 | Union Carbide Corp | |
US3795451A (en) * | 1973-04-24 | 1974-03-05 | Atomic Energy Commission | Rotor for fast analyzer of rotary cuvette type |
US3796239A (en) * | 1971-07-22 | 1974-03-12 | Instrumentation Labor Inc | Dispenser system |
US3798459A (en) * | 1972-10-06 | 1974-03-19 | Atomic Energy Commission | Compact dynamic multistation photometer utilizing disposable cuvette rotor |
US3800161A (en) * | 1972-12-19 | 1974-03-26 | Atomic Energy Commission | Portable dynamic multistation photometer-fluorometer |
US3804533A (en) * | 1972-11-29 | 1974-04-16 | Atomic Energy Commission | Rotor for fluorometric measurements in fast analyzer of rotary |
JPS4971986A (de) * | 1972-09-22 | 1974-07-11 | ||
US3856470A (en) * | 1973-01-10 | 1974-12-24 | Baxter Laboratories Inc | Rotor apparatus |
US3883308A (en) * | 1967-05-12 | 1975-05-13 | Centre Nat Rech Scient | Apparatus for analysing liquid substances likely to form agglutinates |
US3953172A (en) * | 1974-05-10 | 1976-04-27 | Union Carbide Corporation | Method and apparatus for assaying liquid materials |
US3982838A (en) * | 1974-03-12 | 1976-09-28 | The United States Of America As Represented By The United States Energy Research And Development Administration | Compact fast analyzer of rotary cuvette type |
US3988590A (en) * | 1975-04-08 | 1976-10-26 | The United States Of America As Represented By The United States Energy Research And Development Administration | Photomultiplier tube gain regulating system |
US4035156A (en) * | 1977-01-21 | 1977-07-12 | The United States Of America As Represented By The United States Energy Research And Development Administration | Filter type rotor for multistation photometer |
US4123173A (en) * | 1976-06-09 | 1978-10-31 | Electro-Nucleonics, Inc. | Rotatable flexible cuvette arrays |
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Also Published As
Publication number | Publication date |
---|---|
DK134795B (da) | 1977-01-17 |
SE361941B (de) | 1973-11-19 |
NL164391B (nl) | 1980-07-15 |
DE1962267B2 (de) | 1973-11-29 |
DK134795C (de) | 1977-06-06 |
BE742963A (de) | 1970-05-14 |
NL6919034A (de) | 1970-06-22 |
AT297373B (de) | 1972-03-27 |
FR2026450A1 (de) | 1970-09-18 |
DE1962267C3 (de) | 1974-06-27 |
GB1242034A (en) | 1971-08-11 |
DE1962267A1 (de) | 1970-07-09 |
CH502595A (de) | 1971-01-31 |
BR6915271D0 (pt) | 1973-01-18 |
JPS4838274B1 (de) | 1973-11-16 |
ES374546A1 (es) | 1972-01-01 |
NL164391C (nl) | 1980-12-15 |
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