WO1987005323A1 - Sampling of material - Google Patents

Sampling of material Download PDF

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Publication number
WO1987005323A1
WO1987005323A1 PCT/GB1987/000144 GB8700144W WO8705323A1 WO 1987005323 A1 WO1987005323 A1 WO 1987005323A1 GB 8700144 W GB8700144 W GB 8700144W WO 8705323 A1 WO8705323 A1 WO 8705323A1
Authority
WO
WIPO (PCT)
Prior art keywords
sampling
strip
probes
probe
sample
Prior art date
Application number
PCT/GB1987/000144
Other languages
French (fr)
Inventor
William J. Martin
Original Assignee
The University Of Manchester Institute Of Science
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The University Of Manchester Institute Of Science filed Critical The University Of Manchester Institute Of Science
Publication of WO1987005323A1 publication Critical patent/WO1987005323A1/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/02—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by impregnation, e.g. using swabs or loops

Definitions

  • the present invention relates to the sampling of material, particularly but not exclusively microbiological material.
  • Plating out and culture inoculation is a procedure used extensively in life science laboratories.
  • the technique finds wide application in clinical biology as a means of disease identification and treatment, in molecular biology as part of the Sanger DNA sequencing protocol and in pharmaceutical research to screen antibiotics.
  • the method comprises growing microbiological material in, for example, a Petri-dish and the selecting from the resultant growth (or lawn) those colonies or plaques (e.g. of yeasts, fungi, bacteria or virus infected plaques) which are required for further study or amplification.
  • the technique is currently effected manually by experienced personnel who, identify the colonies or plaques of interest, then 'pick-out' these colonies or plaques with individual sampling probes (usually sterile pointed sticks) .
  • a method of obtaining a sample of a material comprising advancing an elongate sampling strip having a plurality of longitudinally spaced sampling probes along a path which successively brings the probes to a sampling position, moving a probe at the sampling position relatively towards the material, moving the probe with collected sample relatively away from the material, and continuing the advance of the strip to bring a successive probe to the sampling position.
  • sampling apparatus comprising a sampling station, an elongate sampling strip extending through the sampling station and having a plurality of longitudinally spaced sampling probes, means for moving a probe at the sampling station relatively towards a material to be sampled, and means for advancing the sampling strip through the apparatus.
  • the strip is preferably of plastics material which may be fed from a roll thereof (e.g. stored in a cassette). The strip may be destroyed (e.g. by incineration) once the samples on the individual probes have been collected for further study or sterilised for re-use.
  • the processing of the samples is effected at a transfer position to which the probes are advanced from the sampling position.
  • the samples may, for example, be transferred to culture tubes or other containers for further growth.
  • the probes carrying the samples are isolated from each other during their passage between the sampling and transfer positions.
  • This may be achieved by an isolating strip which is advanced adjacent with the sampling strip between the two positions and which is formed with a plurality of compartments into each of which one of the probes locate.
  • the method of the invention lends itself readily to automation and may be incorporated into a fully automatic sampling system.
  • a system may comprise automatic visual recognition means for identifying colonies of interest and locating their position in the growth culture. Information from the visual recognition means is then used to control the positioning of the growth culture relative to the sampling strip so that the colony or plaque of interest is proximate the sampling position of the probes.
  • the growth culture may remain stationary and the strip may be positioned above the colony of interest, by movement of sample strip in the horizontal plane.
  • the sampling strip may be incorporated in a mechanical handling robot which effects the sampling procedure. Using such a system a plurality of colonies from each of successive growth cultures supplied to the system may be rapidly sampled and transferred for culture inoculation or other processing.
  • Fig. 1 schematically illustrates one embodiment of the method of the invention
  • Fig. 2 schematically illustrates an automatic sampling system.
  • Fig. 1 schematically illustrates the procedure of the invention for the automatic transfer of selected colonies from a microbiological growth culture in a petri-dish 1 at a sampling station to a culture container 2 at a transfer station.
  • a plastics strip 3 provided with spaced tips 4 (which may be hollow and in the form of pipettes) isfed in the direction of arrow A from a supply (not
  • the strip may, for example be advanced by sprocket wheels engaging in perforations in the edges of the strip or moved by friction drive.
  • the strip 3 moves in the form of a ⁇ around the lower end of a vertically movable plunger 5. It will be seen from Fig. 1 that the width of plunger 5 is so related to the spacing between tips 4 that when one tip projects vertically
  • An identical plunger 6 is provided at the transfer station and a tip 4 projecting vertically below this plunger is referenced in Fig. 1 as
  • the strip 3 is moved in increments (by indexing means not shown) so that the tips 4 stop in the position referenced as 4a and 4b. Additionally, the petri-dish 1 is so moved relative to plunger 5 or vice versa that each successive tip in position 4a is directly above a colony of interest. Downward movement of plunger 5 causes the tip to move down to contact and sample the colony before returning to its upper position by
  • the sample on a tip 4 c at position 4b is transferred to a respective culture container 2 (containing-- suitable liquid) by downward movement of plunger 6.
  • the transfer is effected by virtue of the tip 4 entering the liquid in the container 2 although this may be facilitated by I Q washing means (not shown) directing a washing fluid onto the tip.
  • the strip After leaving the collection station the strip advances to any suitable waste disposal means, e.g. an incinerator (not shown) or to a cleaning system ⁇ for sterilisation before re-use.
  • any suitable waste disposal means e.g. an incinerator (not shown) or to a cleaning system ⁇ for sterilisation before re-use.
  • an isolation strip 7 may be provided. This is a disposable plastics strip
  • the strip has a plurality of cylindrical or other shaped chambers 8 and is moved so that successive tips 4 each locate in one of
  • Means may be provided to allow storage or lengths of the strip between the sampling and transfer stations in a buffer system to allow for
  • Fig. 2 diagrammatically illustrates a complete apparatus for the automatic sampling of selected colonies using the arrangement described with reference to Fig. 1.
  • the cultures to be sampled are grown in sets of rectangular or otherwise shaped containers 10 on a base 11 (5 such containers being shown for each set). These sets are such that when in a vertical stack the base 11 of one set forms 5 a lid for the next lower set.
  • the stacked sets may be positioned on a support 12 which is indexed upwards so that each of the sets may be movedsuccessively from the top of the stack for scanning, sampling and transfer.
  • Other methods may 0 be employed to cover and uncover culture containers such as specially shaped lids, to allow ease of removal for automatic imaging and sampling.
  • a vision system 12 which is such that the position and characteristic (such as colour, shape, size and transparency) of individual colonies in the culture are imaged and recorded. This may be achieved by using a vision system sensitive to small colour and location changes in a Q poor contrast environment. For example, a high resolution CCD matrix or line camera linked to a computer may be employed with background illumination. Intelligent pattern recognition software will be used for characterisation, 5 classification, co-ordinate computation and generation of overall culture statistics.
  • the exact position of colonies of interest may thus be determined and this inforation is used to control the exact position of containers 10 on the 0 X-Y axis in relation to a tip at position 4a so that plunger 5 may be moved downwards to sample the colony of interest in the manner previously described.
  • the plunger 5 may be positioned on the X-Y axis so that a tip at position 4a is vertically 5 above a colony of interest.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

A method of obtaining a sample of a material (particularly a colony or plaque from a microbiological growth culture) comprises advancing an elongate sampling strip (3) having a plurality of longitudinally spaced sampling probes (4) (e.g. in the form of hollow pipettes) along a path which successively brings the probe to a sampling position (4a). A probe at the sampling position is moved relatively towards the material to be sampled, e.g. by a plunger (5), to collect a sample of the material. The probe is then moved away from the material and the advance of the strip is continued to bring a successive probe to the sampling position.

Description

SAMPLING OF MATERIAL
The present invention relates to the sampling of material, particularly but not exclusively microbiological material.
Plating out and culture inoculation is a procedure used extensively in life science laboratories. For example, the technique finds wide application in clinical biology as a means of disease identification and treatment, in molecular biology as part of the Sanger DNA sequencing protocol and in pharmaceutical research to screen antibiotics. Briefly, the method comprises growing microbiological material in, for example, a Petri-dish and the selecting from the resultant growth (or lawn) those colonies or plaques (e.g. of yeasts, fungi, bacteria or virus infected plaques) which are required for further study or amplification. The technique is currently effected manually by experienced personnel who, identify the colonies or plaques of interest, then 'pick-out' these colonies or plaques with individual sampling probes (usually sterile pointed sticks) . The colonies or plaques are then transferred individually to discrete culture tubes or containers in which they are incubated prior to further processing. Many laboratories require hundreds, and possibly thousands, of such operations to be effected daily in a process which is manpower intensive, tedious, repetitive and error prone. Additionally, many cultures of interest may be harmful and consequently there is a health risk with the transfer of such cultures. Furthermore, human operators carry a wide variety of organisms and it is, therefore, difficult to maintain sterile operating conditions when the procedure is being
BAD ORIGINAL carried out manually.
It is an object of the present invention to provide a method of sampling which obviates or mitigates the above entioned disadvantages.
According to a first aspect of the present invention there is provided a method of obtaining a sample of a material, the method comprising advancing an elongate sampling strip having a plurality of longitudinally spaced sampling probes along a path which successively brings the probes to a sampling position, moving a probe at the sampling position relatively towards the material, moving the probe with collected sample relatively away from the material, and continuing the advance of the strip to bring a successive probe to the sampling position.
According to a second aspect of the present invention there is provided sampling apparatus comprising a sampling station, an elongate sampling strip extending through the sampling station and having a plurality of longitudinally spaced sampling probes, means for moving a probe at the sampling station relatively towards a material to be sampled, and means for advancing the sampling strip through the apparatus. The strip is preferably of plastics material which may be fed from a roll thereof (e.g. stored in a cassette). The strip may be destroyed (e.g. by incineration) once the samples on the individual probes have been collected for further study or sterilised for re-use.
Preferably the processing of the samples is effected at a transfer position to which the probes are advanced from the sampling position. At the transfer position, the samples may, for example, be transferred to culture tubes or other containers for further growth.
Preferably also the probes carrying the samples are isolated from each other during their passage between the sampling and transfer positions. This may be achieved by an isolating strip which is advanced adjacent with the sampling strip between the two positions and which is formed with a plurality of compartments into each of which one of the probes locate.
The method of the invention lends itself readily to automation and may be incorporated into a fully automatic sampling system. Such a system may comprise automatic visual recognition means for identifying colonies of interest and locating their position in the growth culture. Information from the visual recognition means is then used to control the positioning of the growth culture relative to the sampling strip so that the colony or plaque of interest is proximate the sampling position of the probes. Alternatively the growth culture may remain stationary and the strip may be positioned above the colony of interest, by movement of sample strip in the horizontal plane. The sampling strip may be incorporated in a mechanical handling robot which effects the sampling procedure. Using such a system a plurality of colonies from each of successive growth cultures supplied to the system may be rapidly sampled and transferred for culture inoculation or other processing. The invention will be further described, by way of example only, with reference to the accompanying drawings; in which:
Fig. 1 schematically illustrates one embodiment of the method of the invention; and Fig. 2 schematically illustrates an automatic sampling system.
Fig. 1 schematically illustrates the procedure of the invention for the automatic transfer of selected colonies from a microbiological growth culture in a petri-dish 1 at a sampling station to a culture container 2 at a transfer station.
A plastics strip 3 provided with spaced tips 4 (which may be hollow and in the form of pipettes) isfed in the direction of arrow A from a supply (not
10 shown) successively through the sampling and transfer stations and then to waste or to a sterilisation area. The strip may, for example be advanced by sprocket wheels engaging in perforations in the edges of the strip or moved by friction drive.
2_5 At the sampling station, the strip 3 moves in the form of a ϋ around the lower end of a vertically movable plunger 5. It will be seen from Fig. 1 that the width of plunger 5 is so related to the spacing between tips 4 that when one tip projects vertically
20 down from the plunger (position 4a) the adjacent tips project horizontally.
An identical plunger 6 is provided at the transfer station and a tip 4 projecting vertically below this plunger is referenced in Fig. 1 as
25 position 4b.
During use of the apparatus, the strip 3 is moved in increments (by indexing means not shown) so that the tips 4 stop in the position referenced as 4a and 4b. Additionally, the petri-dish 1 is so moved relative to plunger 5 or vice versa that each successive tip in position 4a is directly above a colony of interest. Downward movement of plunger 5 causes the tip to move down to contact and sample the colony before returning to its upper position by
__ virtue of upward movement of plunger 5. The strip is advanced so that the tip 4 carrying the sample moves towards the transfer station and a fresh tip moves to position 4a so that the procedure may be repeated.
At the transfer station, the sample on a tip 4 c at position 4b is transferred to a respective culture container 2 (containing-- suitable liquid) by downward movement of plunger 6. The transfer is effected by virtue of the tip 4 entering the liquid in the container 2 although this may be facilitated by IQ washing means (not shown) directing a washing fluid onto the tip.
After leaving the collection station the strip advances to any suitable waste disposal means, e.g. an incinerator (not shown) or to a cleaning system ~ for sterilisation before re-use.
In order to prevent cross-contamination between samples on tips 4 during their passage between the sampling and transfer stations, an isolation strip 7 may be provided. This is a disposable plastics strip
20 fed in the direction of arrow B from a supply (not shown) to disposal means (not shown) or sterilisation means (not shown). The strip has a plurality of cylindrical or other shaped chambers 8 and is moved so that successive tips 4 each locate in one of
25 chambers 8 during their passage between the sampling and transfer stations.
Means may be provided to allow storage or lengths of the strip between the sampling and transfer stations in a buffer system to allow for
30 differences between sampling and transfer rates.
Fig. 2 diagrammatically illustrates a complete apparatus for the automatic sampling of selected colonies using the arrangement described with reference to Fig. 1.
35 In the illustrated apparatus, the cultures to be sampled are grown in sets of rectangular or otherwise shaped containers 10 on a base 11 (5 such containers being shown for each set). These sets are such that when in a vertical stack the base 11 of one set forms 5 a lid for the next lower set. The stacked sets may be positioned on a support 12 which is indexed upwards so that each of the sets may be movedsuccessively from the top of the stack for scanning, sampling and transfer. Other methods may 0 be employed to cover and uncover culture containers such as specially shaped lids, to allow ease of removal for automatic imaging and sampling.
As each set is moved from the stack, its containers 10 are scanned by a vision system 12 which is such that the position and characteristic (such as colour, shape, size and transparency) of individual colonies in the culture are imaged and recorded. This may be achieved by using a vision system sensitive to small colour and location changes in a Q poor contrast environment. For example, a high resolution CCD matrix or line camera linked to a computer may be employed with background illumination. Intelligent pattern recognition software will be used for characterisation, 5 classification, co-ordinate computation and generation of overall culture statistics.
The exact position of colonies of interest may thus be determined and this inforation is used to control the exact position of containers 10 on the 0 X-Y axis in relation to a tip at position 4a so that plunger 5 may be moved downwards to sample the colony of interest in the manner previously described. Alternatively the plunger 5 may be positioned on the X-Y axis so that a tip at position 4a is vertically 5 above a colony of interest.
ORIGINS

Claims

CLAIMS :
1. A method of obtaining a sample of a material, the method comprising advancing an elongate
5 sampling strip having a plurality of longitudinally spaced sampling probes along a path which successively brings the probes to a sampling position, moving a probe at the sampling position relatively towards the material, moving the probe !0 with collected sample relatively away from the material, and continuing the advance of the strip to bring a successive probe to the sampling position.
2. A method as claimed in claim 1 wherein the material to be sampled is a microbiological growth j_5 culture.
3. A method as claimed in claim 1 wherein the probes are hollow.
4. A method as claimed in claim 1 wherein at the sampling position the sampling strip extends
20 around the bottom of a reciprocable plunger or the like, whereby downward movement of the plunger moves the lower portion of the strip and a probe carried thereon towards the material to be sampled.
5. A method as claimed in claim 1 wherein the 25 probes with the collected sample are advanced to a transfer position at which the samples are transferred from the probes.
6. A method as claimed in claim 3 wherein the probes are isolated from each other during their
30 passage from the sampling position to the transfer position.
7. A method as claimed in claim 6 wherein said isolation is effected by means of an isolating strip having a plurality of longitudinally spaced
35 compartments, said isolating strip being advanced adjacent the sampling strip between the sampling and transfer positions with each of said probes locating in a respective compartment.
8. A method as claimed in claim 5 wherein at the transfer position the sampling strip extends around the bottom of a reciprocable plunger or the like whereby downward movement of the plunger moves the lower portion of the strip and a probe carried thereon towards a vessel to which the sample is to be transferred.
9. A method as claimed in claim 1 wherein the sampling strip is of plastics material.
10. A method as claimed in claim 1 wherein the sampling strip is disposable.
11. A method as claimed in claim 1 wherein the sampling strip is stored in a cassette.
12. Sampling apparatus comprising a sampling station, an elongate sampling strip extending through the sampling station and having a plurality of longitudinally spaced sampling probes, means for moving a probe at the sampling station relatively towards a material to be sampled, and means for advancing the sampling strip through the apparatus.
13. Apparatus as claimed in claim 12 wherein the means for moving the probe relatively towards a material to be sampled is a reciprocable plunger or the like around the Dottom of which the sampling strip locates.
14. Apparatus as claimed in- claim 12 additionally comprising a transfer station, said transfer station having means for moving a probe located at a transfer position towards a vessel to which a sample is to be transferred.
15. Apparatus as claimed in claim 14 wherein the means for moving the probe relatively towards a ~ vessel is a reciprocable plunger or the like around the bottom of which the sampling strip locates.
16. Apparatus as claimed in claim 14 provided with means for isolating probes between the sampling and transfer station from each other.
17. Apparatus as claimed in claim 16 wherein the isolating means comprises an isolating strip having a plurality of longitudinally spaced compartments, said isolating strip being located adjacent said sampling strip whereby the probes each locates in a respective compartment.
18. Apparatus as claimed in claim 10 comprising visual recognition means, and means for controlling the positioning of the sample relative to the sampling strip in accordance with information from the visual recognition means.
19. Apparatus as claimed in claim 10 wherein the visual recognition means is capable of identifying microbiological colonies.
20. Apparatus as claimed in claim 10 wherein the sampling strip is incorporated in a mechanical handling robot.
21. An elongate sampling strip having a plurality of longitudinally spaced sampling probes.
PCT/GB1987/000144 1986-03-01 1987-03-02 Sampling of material WO1987005323A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB868605140A GB8605140D0 (en) 1986-03-01 1986-03-01 Sampling of microbiological material
GB8605140 1986-03-01

Publications (1)

Publication Number Publication Date
WO1987005323A1 true WO1987005323A1 (en) 1987-09-11

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PCT/GB1987/000144 WO1987005323A1 (en) 1986-03-01 1987-03-02 Sampling of material

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GB (1) GB8605140D0 (en)
WO (1) WO1987005323A1 (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5063791A (en) * 1987-07-31 1991-11-12 National Research Development Corporation Sampling of material
WO1992012233A1 (en) * 1991-01-11 1992-07-23 Medical Research Council Transfer of biological samples
WO1999066023A2 (en) * 1998-06-15 1999-12-23 Gesellschaft Fuer Biotechnologische Forschung Mbh (Gbf) Device for taking up and transferring biological samples
DE10209348A1 (en) * 2001-08-06 2003-03-06 Grohmann Engineering Gmbh Transfer of mass laboratory liquid samples involves moving to test instrument using one-trip tape-mounted transfer tool

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3455788A (en) * 1966-11-15 1969-07-15 Lever Brothers Ltd Multiple inoculation device
DE2351617A1 (en) * 1973-10-15 1975-04-24 Saul Kaye Quantitative microorganism concn determining appts - contg a number of containers having different volumetric capacities and exposed simultaneously to sampling
DE3146101A1 (en) * 1980-11-21 1982-06-16 Michael Peter Etobicoke Ontario Entis "DEVICE FOR TRANSMITTING MICROORGANISMS"

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3455788A (en) * 1966-11-15 1969-07-15 Lever Brothers Ltd Multiple inoculation device
DE2351617A1 (en) * 1973-10-15 1975-04-24 Saul Kaye Quantitative microorganism concn determining appts - contg a number of containers having different volumetric capacities and exposed simultaneously to sampling
DE3146101A1 (en) * 1980-11-21 1982-06-16 Michael Peter Etobicoke Ontario Entis "DEVICE FOR TRANSMITTING MICROORGANISMS"

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5063791A (en) * 1987-07-31 1991-11-12 National Research Development Corporation Sampling of material
WO1992012233A1 (en) * 1991-01-11 1992-07-23 Medical Research Council Transfer of biological samples
WO1999066023A2 (en) * 1998-06-15 1999-12-23 Gesellschaft Fuer Biotechnologische Forschung Mbh (Gbf) Device for taking up and transferring biological samples
DE19826244A1 (en) * 1998-06-15 1999-12-23 Biotechnolog Forschung Gmbh Device for recording and transferring biological samples
WO1999066023A3 (en) * 1998-06-15 2000-10-26 Biotechnolog Forschung Gmbh Device for taking up and transferring biological samples
DE19826244C2 (en) * 1998-06-15 2003-06-26 Biotechnolog Forschung Gmbh Device for recording and transferring biological samples
DE10209348A1 (en) * 2001-08-06 2003-03-06 Grohmann Engineering Gmbh Transfer of mass laboratory liquid samples involves moving to test instrument using one-trip tape-mounted transfer tool

Also Published As

Publication number Publication date
JPS63502721A (en) 1988-10-13
GB8605140D0 (en) 1986-04-09

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