Method and apparatus for signalling the introduction of chemical reaction components into a chemical analyzing system

• US 4,101,276 A
• Filed: 06/02/1976
• Issued: 07/18/1978
• Est. Priority Date: 06/02/1976
• Status: Expired due to Term

• Current Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Original Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Inventors: Anderson, Robert J.
• Primary Examiner(s): Reese, Robert M.
• Application Number:
  US05/692,159
• Time in Patent Office: 776 Days
• Field of Search: 23/230 B, 23/253 R, 356/39, 356/73, 73/64.1, 250/461, 250/461 B
• US Class Current: 436/536
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 31/00 Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures

Abstract

A system for chemically analyzing antigens or antibodies by initiating an antigen-antibody reaction in a reaction zone and measuring the light scattered by a precipitate produced by the reaction to determine the antigen or antibody concentration. In order to signal the start of the reaction, a fluorescent tagging substance is included with at least the last antigen or antibody reaction component introduced into the reaction zone. The fluorescent tagging substance does not enter into the antigen-antibody reaction and is selected to emit light in a bandwidth spectrally separated from the light scattered by the precipitate. The reaction zone is monitored for the presence of the tagging substance, and when the substance is detected a trigger signal is generated to signal the start of the reaction. By including a tagging substance with each component of the reaction and by monitoring the reaction zone for the presence of each tagging substance, a trigger signal is generated only if each reaction component is present.

Claims

1. In a method of combining components of a chemical reaction which includes the steps of introducing the reaction components into a reaction zone to initiate the chemical reaction and measuring a characteristic of the reaction, the improved method of signalling the start of the chemical reaction comprising the steps of:

prior to initiating the chemical reaction adding a tagging substance to at least the reaction component last introduced into the reaction zone, the tagging substance remaining chemically isolated from the reaction; and

monitoring the reaction zone for the presence of the tagging substance and upon detecting the tagging substance generating a signal indicating the start of the chemical reaction.

2. The method of claim 1 wherein the tagging substance is added to at least an antigen or antibody component of an antigen-antibody chemical reaction.

3. The method of claim 1 including the steps of:

adding the tagging substance to a second reaction component in addition to said last introduced reaction component; and

monitoring the reaction zone for the presence of each tagging substance and upon detection of both generating the signal indicating the start of the chemical reaction.

4. The method of claim 3 wherein the tagging substance is added to the first and second reaction components in different relative amounts and the reaction zone is monitored for the presence of the unique combination of the combined different relative amounts of the tagging substance.

5. The method of claim 1 wherein:

the tagging substance is a fluorescent substance; and

the step of monitoring includes detecting light emitted by the fluorescent substance to signal the start of the chemical reaction.

6. The method of claim 5 wherein the fluorescent substance is oxazine-1-perchlorate.

7. The method of claim 5 wherein:

the step of measuring a characteristic of the reaction measures an optical characteristic thereof in a first spectral band;

the fluorescent tagging substance is selected to emit light in a third spectral band separate from said first spectral band; and

the step of monitoring includes detecting light in the third spectral band.

8. The method of claim 7 wherein the fluorescent tagging substance is further selected to absorb light in a second spectral band separate from the first and third spectral bands.

9. The method of claim 8 wherein the fluorescent tagging substance is added to a component of an antigen-antibody chemical reaction, and said optical characteristic is light scattered by a precipitate formed during the reaction.

10. An electrochemical system for detecting the combining of components of a chemical reaction comprising:

a reaction chamber for serially receiving a plurality of said components for a chemical reaction within the chamber;

means operatively coupled to said chamber for measuring a characteristic of said chemical reaction;

a tagging substance in at least a last one of said components received in said chamber and chemically isolated from said reaction;

separate means operatively coupled to said chamber and insensitive to said characteristic of said chemical reaction for detecting said tagging substance;

triggering means responsive to a detection of said tagging substance for generating an electrical signal indicative of a combining of said components and a start of said chemical reaction;

electrical signal responsive means; and

means for supplying said electrical signal to said electrical signal responsive means.

11. The system of claim 10 wherein the measuring means measures an optical characteristic of the reaction;

the tagging substance is a fluorescent substance which fluoresces in a spectral band separate from that of the optical characteristic of the reaction; and

the detecting means detects fluorescent light issuing from said tagging substance to the exclusion of said optical characteristic.

12. The system of claim 10 wherein said electrical signal responsive means includes a clock;

and

said supplying means supplies said electrical signal as a triggering input to start said clock.

13. A nephelometric chemical analyzer comprising:

a reaction chamber for receiving components for a chemical reaction within the chamber;

excitation means for directing a beam of light having first and second separate spectral bands into the reaction chamber;

detection means for detecting light scattered by the contents of the reaction chamber within said first spectral band to measure a scattered light characteristic of the reaction for nephelometric analysis;

a fluorescent tagging substance in at least a last one of said components introduced into the reaction chamber and chemically isolated from the chemical reaction, said fluorescent tagging substance absorbing light in said second spectral band and emitting light in a third spectral band separate from each of said first and second spectral bands;

separate means operatively coupled to said reaction chamber and insensitive to said scattered light characteristic in said first spectral band for detecting fluorescent light issuing from the fluorescent tagging substance in said third spectral band;

triggering means responsive to a detection of the tagging substance for generating an electrical signal indicative of a combining of said components and start of said chemical reaction;

a clock; and

means for supplying said electrical signal as a triggering input to start said clock.

14. The analyzer of claim 13 wherein said excitation means includes a source of light and primary filter means for intercepting light from the light source and for passing light in said first and second spectral bands into the reaction chamber while rejecting light in said third spectral band.

15. The analyzer of claim 13 wherein said detection means includes a detector, and secondary filter means for intercepting scattered light from the reaction chamber and for passing light within said first spectral band to said detector while rejecting light in said second and third spectral bands.

16. The apparatus of claim 13 wherein said separate means includes a detector, and filter means for intercepting light from the reaction chamber and for passing light in said third spectral band to said detector while rejecting light in said first and second spectral bands.