Patent US3902971A successfully refreshed from Google Patents.
US3902971A
Expired / Lapsed
US
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Biological detecting method and apparatus
Assignee: Akzona Inc
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Filing: 1965-05-20
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Granted: 1975-09-02
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Expires: 1985-05-20
Abstract
Abstract This disclosure describes apparatus and the method for continuously sampling either gaseous or liquid media such as atmosphere or water to detect the presence or absence of specific pathogenic micro-organisms. The system as described employs collection apparatus which entrains particles of microorganism size in continuous flowing stream of water. A suspension of fluorescently tagged antibodies is injected into the flowing stream and thereby maintained in contact for a sufficient period of time for the specific antibodies to attach to any of its corresponding organism in the collection stream. The resultant mixture of the two streams is passed through a continuous filter having pore size selected to hold on its surface reacted antigenantibody complexes while passing the remainder of the affluent. After washing to remove extraneous residue, the surface of the filter is exposed to light of a wavelength which excites the fluorescently tagged antibodies if resident on the tape, and the number of such fluorescent particles appearing in the field of a scanned field is counted. In the absence of any fluorescent particles, the original collection stream is known to be free of the particular microorganism. The disclosure describes specific antibody-antigen complexes and their recommended fluorescent dye and exciting wavelength. A wide band preamplifier for detecting the signals on the system is similarly disclosed.
Inventors
- 👤 James C Fletcher
- 👤 Charles E Hendrix
- 👤 Scott B Kilner
- 👤 Frank W Lehan
- 👤 Paul L Magill
- 👤 Eli Mishuck
- 👤 Jr William A Perkins
- 👤 Merton A Robinson
- 👤 Keith H Sweeny
- 👤 Richard W Tew
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 07:30
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/863; Y10S435/866; Y10S435/871
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
103 words
A SYSTEM FOR DETECTING MICRO-ORGANISMS COMPRISING: MEANS FOR SAMPLING A FLUID MEDIUM TO BE MONTORED FOR SEPECIFIC MICRO-ORGANISMS, A SOURCE OF ANTISERUM CONTAINING TAGGED ANTIBODIES CAPABLE OF REACTING WITH THE SPECIFIC MICRO-ORGANISMS, MEANS FOR INTRODUCING THE ANTISERUM INTO A STREAM OF THE SAMPLED FLUID MEDIUM, MEANS FOR MAINTAINING THE MIXED SAMPLED AND ANTISERUM STREAM AT A TEMPERATURE FOR A PERIOD OF TIME SUFFIEIENT FOR THE MICRO-ORGANISMS PRESENT AND ANTIBODIES TO REACT, FILTER MEANS FOR SEPARTING ON THE SURFACE THEREOF PARTICULATE MATERIAL INCLUDING REACTED MICRO-ORGANISMS, MEANS FOR DISPENSING THE STREAM OF MIXED SAMPLED FLUID MEDIUM AND ANTISERUM ONTO THE SURFACE OF THE FILTER MEANS,
Claim 2
Independent Claim
103 words
A SYSTEM FOR DETECTING MICRO-ORGANISMS COMPRISING: MEANS FOR SAMPLING A FLUID MEDIUM TO BE MONTORED FOR SEPECIFIC MICRO-ORGANISMS, A SOURCE OF ANTISERUM CONTAINING TAGGED ANTIBODIES CAPABLE OF REACTING WITH THE SPECIFIC MICRO-ORGANISMS, MEANS FOR INTRODUCING THE ANTISERUM INTO A STREAM OF THE SAMPLED FLUID MEDIUM, MEANS FOR MAINTAINING THE MIXED SAMPLED AND ANTISERUM STREAM AT A TEMPERATURE FOR A PERIOD OF TIME SUFFIEIENT FOR THE MICRO-ORGANISMS PRESENT AND ANTIBODIES TO REACT, FILTER MEANS FOR SEPARTING ON THE SURFACE THEREOF PARTICULATE MATERIAL INCLUDING REACTED MICRO-ORGANISMS, MEANS FOR DISPENSING THE STREAM OF MIXED SAMPLED FLUID MEDIUM AND ANTISERUM ONTO THE SURFACE OF THE FILTER MEANS,
Claim 3
Dependent Claim
158 words
A detection system for air-borne micro-organisms comprising: an air sampler; means for separating from the air to be sampled particulate material larger than the mean dimensions of the micro-organisms to be detected; means for aspirating a spray of water into the sampled air to entrain therein residual particulate material including micro-organisms; a collector for the aspirated water and micro-organisms, including a conduit for producing a continuous stream of micro-organism-carrying water; means for introducing tagged antiserum containing antibodies specific to the micro-organisms into the stream of micro-organism-carrying water; means for maintaining the mixed sample and antiserum stream at a temperature for a period of time sufficient for the micro-organisms present and antibodies to react; a microporous filter having pore size to retain micro-organisms on the surface thereof; means for passing the collected water through the microporous filter to separate the reacted antibody-micro-organism complexes from the residual fluid; and means for scanning the Surface of the filter for tagged organisms.
Claim 4
Dependent Claim
158 words
A detection system for air-borne micro-organisms comprising: an air sampler; means for separating from the air to be sampled particulate material larger than the mean dimensions of the micro-organisms to be detected; means for aspirating a spray of water into the sampled air to entrain therein residual particulate material including micro-organisms; a collector for the aspirated water and micro-organisms, including a conduit for producing a continuous stream of micro-organism-carrying water; means for introducing tagged antiserum containing antibodies specific to the micro-organisms into the stream of micro-organism-carrying water; means for maintaining the mixed sample and antiserum stream at a temperature for a period of time sufficient for the micro-organisms present and antibodies to react; a microporous filter having pore size to retain micro-organisms on the surface thereof; means for passing the collected water through the microporous filter to separate the reacted antibody-micro-organism complexes from the residual fluid; and means for scanning the Surface of the filter for tagged organisms.
Claim 5
Independent Claim
150 words
A system for monitoring atmosphere for specific micro-organisms, comprising: means for sampling the atmosphere to be monitored; means for separating particulate material above the size of such micro-organisms; means for entraining the residual particulate material including micro-organisms into a stream of an aqueous solution; a source of antiserum containing tagged antibodies capable of reacting with the micro-organisms to be detected; means for continuously introducing the antiserum into the stream of aqueous solution; means for maintaining the mixed stream of solution and antiserum at a temperature for a period of time sufficient for micro-organisms present to react with tagged antibodies; means for depositing the reacted antibody-micro-organism complex from the stream on the surface of a porous filter tape; means for illuminating discrete areas of the surface of the filter tape with radiation causing the tagged organisms to be observable; and means for counting the observable tagged organisms in each discrete area.
Claim 6
Independent Claim
150 words
A system for monitoring atmosphere for specific micro-organisms, comprising: means for sampling the atmosphere to be monitored; means for separating particulate material above the size of such micro-organisms; means for entraining the residual particulate material including micro-organisms into a stream of an aqueous solution; a source of antiserum containing tagged antibodies capable of reacting with the micro-organisms to be detected; means for continuously introducing the antiserum into the stream of aqueous solution; means for maintaining the mixed stream of solution and antiserum at a temperature for a period of time sufficient for micro-organisms present to react with tagged antibodies; means for depositing the reacted antibody-micro-organism complex from the stream on the surface of a porous filter tape; means for illuminating discrete areas of the surface of the filter tape with radiation causing the tagged organisms to be observable; and means for counting the observable tagged organisms in each discrete area.
Claim 7
Dependent Claim
185 words
A detection system for micro-organisms comprising: means for sampling a fluid medium to be monitored for particular micro-organisms; means for entraining particulate material of the sample of the medium in an aqueous carrier stream; a source of tagged antiserum containing antibodies specific to the micro-organisms to be detected; means for continuously introducing the tagged antiserum into the aqueous carrier stream; conduit means for transporting the mixed streams while maintaining the particulate-carrying aqueous carrier and antiserum at a temperature for sufficient time for the antibodies and micro-organisms to react; a filter tape having a pore size retaining reacted micro-organism-antibody complexes on the surface thereof while passing unreacted antiserum; means for passing the aqueous carrier and antiserum stream through the filter tape; means for advancing the filter tape to present successive areas of the tape of the aqueous carrier; flying spot scanning means positioned to illuminate localized areas of the surface of the filter tape with radiation causing the tagged antibodies of each antibody-micro-organism complex to be visible; means for detecting visible complexes as illuminated by the flying spot scanning means; and means for counting visible complexes.
Claim 8
Dependent Claim
185 words
A detection system for micro-organisms comprising: means for sampling a fluid medium to be monitored for particular micro-organisms; means for entraining particulate material of the sample of the medium in an aqueous carrier stream; a source of tagged antiserum containing antibodies specific to the micro-organisms to be detected; means for continuously introducing the tagged antiserum into the aqueous carrier stream; conduit means for transporting the mixed streams while maintaining the particulate-carrying aqueous carrier and antiserum at a temperature for sufficient time for the antibodies and micro-organisms to react; a filter tape having a pore size retaining reacted micro-organism-antibody complexes on the surface thereof while passing unreacted antiserum; means for passing the aqueous carrier and antiserum stream through the filter tape; means for advancing the filter tape to present successive areas of the tape of the aqueous carrier; flying spot scanning means positioned to illuminate localized areas of the surface of the filter tape with radiation causing the tagged antibodies of each antibody-micro-organism complex to be visible; means for detecting visible complexes as illuminated by the flying spot scanning means; and means for counting visible complexes.
Claim 9
Dependent Claim
55 words
The combination in accordance with claim 4 wherein both the filter means is advanced and the flying spot scanning means illuminates sequential areas of the filter means at rates related to the fluid medium sampling rate whereby the count of visible antibodies per unit of time of said counting means indicates the concentration of micro-organisms.
Claim 10
Dependent Claim
82 words
Apparatus for counting micro-organisms comprising: means for injecting antiserum containing tagged antibodies specific to the micro-organism into a fluid stream containing the micro-organisms to be counted; conduit means for transporting the fluids while maintaining the mixed antiserum and fluid stream at reaction temperature for a period sufficient to allow the antibody-antigen reaction to occur; filter means at the outlet of said conduit means for separating reacted antibody-micro-organism complexes from the fluid medium and unreacted antiserum; and means for counting the tagged antibodies.
Claim 11
Dependent Claim
82 words
Apparatus for counting micro-organisms comprising: means for injecting antiserum containing tagged antibodies specific to the micro-organism into a fluid stream containing the micro-organisms to be counted; conduit means for transporting the fluids while maintaining the mixed antiserum and fluid stream at reaction temperature for a period sufficient to allow the antibody-antigen reaction to occur; filter means at the outlet of said conduit means for separating reacted antibody-micro-organism complexes from the fluid medium and unreacted antiserum; and means for counting the tagged antibodies.
Claim 12
Dependent Claim
31 words
The combination in accordance with claim 6 wherein said counting means includes a source of illumination causing the tagged antibodies to be visible and photo-responsive means for counting the visible antibodies.
Claim 13
Dependent Claim
32 words
The combination in accordance with claim 6 wherein the antiserum contains antibodies tagged with fluorescent dye and the source of illumination produces radiation of wavelength producing fluorescent emission from the tagged antibodies.
Claim 14
Dependent Claim
79 words
Apparatus for identifying specific micro-organisms in a fluid medium comprising: a conduit for caRrying the fluid to be sampled; means for injecting into the fluid medium carrying conduit a stream of antiserum containing tagged antibodies specific to the micro-organisms to be identified; filter means for separating from the fluid medium the particulate material of the order of size of the micro-organism while retaining in the fluid medium unreacted antiserum; and means for examining the separated material for tagged antibodies.
Claim 15
Dependent Claim
79 words
Apparatus for identifying specific micro-organisms in a fluid medium comprising: a conduit for caRrying the fluid to be sampled; means for injecting into the fluid medium carrying conduit a stream of antiserum containing tagged antibodies specific to the micro-organisms to be identified; filter means for separating from the fluid medium the particulate material of the order of size of the micro-organism while retaining in the fluid medium unreacted antiserum; and means for examining the separated material for tagged antibodies.
Claim 16
Independent Claim
99 words
A method of detecting micro-organisms comprising the steps of: extracting a sample stream of a fluid medium to be monitored for micro-organisms; mixing the sample stream with a stream of tagged antiserum containing antibodies specific to the micro-organisms to be detected; maintaining the mixed streams at reaction temperature for sufficient period of time for the antibodies and micro-organisms to react; depositing the reacted micro-organism-antibody complexes on a porous carrier capable of retaining on the surface thereof micro-organisms while passing unreacted antiserum; illuminating the complexes with radiation causing the tagged antibodies to be visible; and observing the visible antibody-micro-organism complexes.
Claim 17
Independent Claim
99 words
A method of detecting micro-organisms comprising the steps of: extracting a sample stream of a fluid medium to be monitored for micro-organisms; mixing the sample stream with a stream of tagged antiserum containing antibodies specific to the micro-organisms to be detected; maintaining the mixed streams at reaction temperature for sufficient period of time for the antibodies and micro-organisms to react; depositing the reacted micro-organism-antibody complexes on a porous carrier capable of retaining on the surface thereof micro-organisms while passing unreacted antiserum; illuminating the complexes with radiation causing the tagged antibodies to be visible; and observing the visible antibody-micro-organism complexes.
Claim 18
Independent Claim
102 words
A method of detecting micro-organisms comprising the steps of: extracting a sample of a fluid medium to be monitored for micro-organisms; mixing the sample with a fluid antiserum containing antibodies tagged with fluorescent material; maintaining the sample and antiserum at a temperature of 25* C for a period of one to two minutes to allow the fluorescent-tagged antibodies and micro-organisms to react; passing the mixed sample and fluid antiserum through a filter to separate the micro-organisms and residual particulate material from the unreacted antibodies and filtrate; illuminating the filter with radiation causing the tagged antibodies to fluoresce; and counting the fluorescent particles.
Claim 19
Independent Claim
102 words
A method of detecting micro-organisms comprising the steps of: extracting a sample of a fluid medium to be monitored for micro-organisms; mixing the sample with a fluid antiserum containing antibodies tagged with fluorescent material; maintaining the sample and antiserum at a temperature of 25* C for a period of one to two minutes to allow the fluorescent-tagged antibodies and micro-organisms to react; passing the mixed sample and fluid antiserum through a filter to separate the micro-organisms and residual particulate material from the unreacted antibodies and filtrate; illuminating the filter with radiation causing the tagged antibodies to fluoresce; and counting the fluorescent particles.
Claim 20
Independent Claim
96 words
A method of detecting atmosphere-borne micro-organisms comprising the steps of: entraining the particulate material and micro-organisms of a sample of atmosphere in a stream of water; mixing the stream of water with a stream of fluid antiserum containing fluorescent-tagged antibodies; maintaining the mixed stream and antiserum at a temperature for a sufficient period of time for the tagged antibodies and micro-organisms to react; passing the resultant fluids through a filter retaining particulate material including the reacted antibody-micro-organism complexes; illuminating the surface of the filter to cause the tagged antibodies to fluoresce; and counting the fluorescing particles.
Claim 21
Independent Claim
96 words
A method of detecting atmosphere-borne micro-organisms comprising the steps of: entraining the particulate material and micro-organisms of a sample of atmosphere in a stream of water; mixing the stream of water with a stream of fluid antiserum containing fluorescent-tagged antibodies; maintaining the mixed stream and antiserum at a temperature for a sufficient period of time for the tagged antibodies and micro-organisms to react; passing the resultant fluids through a filter retaining particulate material including the reacted antibody-micro-organism complexes; illuminating the surface of the filter to cause the tagged antibodies to fluoresce; and counting the fluorescing particles.
Claim 22
Independent Claim
128 words
A method of detecting the concentration of particular micro-organisms in a fluid medium comprising the steps of: sampling the fluid medium at a predetermined rate; mixing the stream of sampled medium with a stream of antiserum containing tagged antibodies specific to the micro-organisms to be detected; maintaining the mixed streams at a temperature for a period of time sufficient for the tagged antibodies and micro-organisms present to react; passing the mixed streams at a constant rate through discrete areas of filter medium having pore size which retains micro-organisms and passes the fluid filtrate; illuminating local areas of the surface of the filter medium with radiation causing the tagged micro-organisms to be visible; and counting the number of visible micro-organisms in each discrete area to indicate the micro-organism concentration.
Claim 23
Independent Claim
128 words
A method of detecting the concentration of particular micro-organisms in a fluid medium comprising the steps of: sampling the fluid medium at a predetermined rate; mixing the stream of sampled medium with a stream of antiserum containing tagged antibodies specific to the micro-organisms to be detected; maintaining the mixed streams at a temperature for a period of time sufficient for the tagged antibodies and micro-organisms present to react; passing the mixed streams at a constant rate through discrete areas of filter medium having pore size which retains micro-organisms and passes the fluid filtrate; illuminating local areas of the surface of the filter medium with radiation causing the tagged micro-organisms to be visible; and counting the number of visible micro-organisms in each discrete area to indicate the micro-organism concentration.
Claim 24
Independent Claim
152 words
A method of detecting the concentration of particular micro-organisms in the atmosphere comprising the steps of: sampling the atmosphere at a predetermined constant rate; separating particulate material of size greater than the mean size of the micro-organism to be detected; entraining the residual particulate material including micro-organisms in a fluid medium; mixing at a constant rate a stream of the fluid medium with a stream of antiserum containing tagged antibodies specific to the micro-organisms to be detected; maintaining the mixed streams at a temperature for a period of time sufficient for the tagged antibodies and micro-organisms to react; passing the resultant stream through a porous filter having a pore size which passes the filtrate and retains the reacted micro-organism-antibody complexes on the surface thereof; illuminating in sequence local areas of the filter surface of size approximating the nominal diameter of a single such complex; and counting the number of discrete complexes visible.
Claim 25
Independent Claim
152 words
A method of detecting the concentration of particular micro-organisms in the atmosphere comprising the steps of: sampling the atmosphere at a predetermined constant rate; separating particulate material of size greater than the mean size of the micro-organism to be detected; entraining the residual particulate material including micro-organisms in a fluid medium; mixing at a constant rate a stream of the fluid medium with a stream of antiserum containing tagged antibodies specific to the micro-organisms to be detected; maintaining the mixed streams at a temperature for a period of time sufficient for the tagged antibodies and micro-organisms to react; passing the resultant stream through a porous filter having a pore size which passes the filtrate and retains the reacted micro-organism-antibody complexes on the surface thereof; illuminating in sequence local areas of the filter surface of size approximating the nominal diameter of a single such complex; and counting the number of discrete complexes visible.
Claim 26
Dependent Claim
39 words
The method in accordance with claim 14 wherein the mixed streams are deposited on progressive areas of a porous filter whereby the number of complexes detected per unit area is an indication of the micro-organism concentration of the atmosphere.
Claim 27
Dependent Claim
31 words
The method in accordance with claim 14 wherein the tagged complexes are illuminated with radiation of wavelength causing the tagged antibody to exhibit distinct emission as compared with background particulate material.
Claim 28
Dependent Claim
28 words
The method in accordance with claim 16 wherein the complexes are tagged with fluorescein isothiocyanate and are illuminated with radiation in the blue-violet range of the visible spectrum.
Claim 29
Independent Claim
56 words
A method of staining micro-organisms comprising: entraining micro-organisms to be stained in a fluid medium; mixing the medium with an antiserum containing stained antibodies specific to the micro-organism to be detected; and maintaining the mixed fluid medium and antiserum at a temperature for a period of time sufficient for the micro-organisms and tagged antibodies to react.
Claim 30
Independent Claim
56 words
A method of staining micro-organisms comprising: entraining micro-organisms to be stained in a fluid medium; mixing the medium with an antiserum containing stained antibodies specific to the micro-organism to be detected; and maintaining the mixed fluid medium and antiserum at a temperature for a period of time sufficient for the micro-organisms and tagged antibodies to react.
Claim 31
Independent Claim
56 words
A method of staining micro-organisms comprising: entraining micro-organisms to be stained in an aqueous stream; mixing the stream with a stream of antiserum containing stained antibodies; maintaining the mixed streams at a temperature for a period of time sufficient for the micro-organisms and antibodies to react; and separating the reacted complexes from the residual unreacted antiserum.
Claim 32
Independent Claim
56 words
A method of staining micro-organisms comprising: entraining micro-organisms to be stained in an aqueous stream; mixing the stream with a stream of antiserum containing stained antibodies; maintaining the mixed streams at a temperature for a period of time sufficient for the micro-organisms and antibodies to react; and separating the reacted complexes from the residual unreacted antiserum.
References Cited (Backward Citations)
US3128239A
Cited
US3129144A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1978-09-01 | 1979-03-03 | 1979-09-02 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-09-01 | 1983-03-03 | 1983-09-02 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-09-01 | 1987-03-03 | 1987-09-02 | $7700.00 | Window Closed / Lapsed |