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WO1996037301A1 - Ameliorations apportees a une technique de filtration - Google Patents

Ameliorations apportees a une technique de filtration Download PDF

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Publication number
WO1996037301A1
WO1996037301A1 PCT/GB1996/001140 GB9601140W WO9637301A1 WO 1996037301 A1 WO1996037301 A1 WO 1996037301A1 GB 9601140 W GB9601140 W GB 9601140W WO 9637301 A1 WO9637301 A1 WO 9637301A1
Authority
WO
WIPO (PCT)
Prior art keywords
disc
filtration device
tubular body
collection vessel
membrane
Prior art date
Application number
PCT/GB1996/001140
Other languages
English (en)
Inventor
Gary James Clements
Richard Michael Ireland
Original Assignee
Haemocell Plc
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 Haemocell Plc filed Critical Haemocell Plc
Publication of WO1996037301A1 publication Critical patent/WO1996037301A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/14—Ultrafiltration; Microfiltration
    • B01D61/18—Apparatus therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures

Definitions

  • a supposed dispersion of cellular material in, say, 500 ml of a liquid medium is passed through a filter membrane so that any cellular material present is retained by the membrane.
  • the membrane is then placed in a Petri dish, nutrient is added, and the dish is incubated for a period of time to produce a culture of the material.
  • a lysing solution is added and thereafter a sample of say 0.2 ml of the lysate is extracted from the dish by means of a pipette and subjected to analysis, for example by being deposited into a reagent in a luminometer cuvette.
  • This technique suffers the disadvantages that there are many stages during which the material is exposed to the atmosphere and may be contaminated by exogenous ATP, and in that the final sample used for analysis is only an aliquot of the available material.
  • the present invention provides a filtration technique for use in the preparation of a sample of concentrated cellular material in conjunction with rapid micro ⁇ biological quality assurance, or with non-microbiological applications, such as in mammalian or insect cell tissue culture, or in the analysis of certain clinical samples.
  • a filtration device comprises a tubular body having at one end an inlet which is arranged to be sealed to the outlet of a source of a dispersion of the cellular material which can be put under pressure, at its other end an outlet, and between the inlet and the outlet a seating; a disc which is perforated between its opposed major faces and which is provided over one of its faces with a filter membrane, the disc being held on the seating with the membrane both facing the one end and closing the passage through the body between the inlet and the outlet so as, in use, to filter out and retain thereon cellular material in a dispersion flowing through the tubular body from the inlet; and means for releasing the disc whereby in use, the disc with the retained cellular material on the membrane is free to pass through the outlet into a collection vessel for subsequent analysis.
  • a method of evaluating a cellular material comprises forcing a dispersion of a cellular material from a source through a tubular body the passage through which is closed by a filter membrane carried by a supporting disc which is held in the body whereby the cellular material is retained on the membrane; and releasing the disc so that the disc and membrane pass from the body into a collection vessel, and subsequently carrying out an appropriate evaluation.
  • the source of the dispersion of the cellular material to be forced through the tubular body of the new filtration device may be a conventional syringe.
  • the syringe outlet may be conventionally tapered and either held, or secured as an interference fit, in the inlet of the tubular body, but is preferably connectable to it positively by means, e.g., of a Luer lock.
  • the use of a syringe is particularly appropriate if the filtration is to follow primary filtration in a system in which the recovered concentrated cellular material is collected in a vessel formed by a syringe. The same syringe containing the same concentration can then be removed from the primary filtration system and directly fitted to the tubular body of the secondary filtration device with minimum atmospheric exposure.
  • the vessel which is used to collect the disc, filter membrane and retained material, following the secondary filtration in the new device may be a reaction vessel for analytical equipment, such as an impedance device; or an assembly for microscopic analysis.
  • the vessel will be a tube, such as a luminometer cuvette, in the typical case in which the evaluation is to be carried out by bioluminescence or epifluorescence.
  • the shape and dimensions of the disc, and the internal dimensions of the cuvette or other tube are preferably such that the disc cannot turn over as it falls into the tube so that the membrane-covered surface of the disc remains uppermost, i.e. facing the open end of the tube.
  • the peripheral surface of the disc is cylindrical with a diameter sufficiently smaller than the internal diameter of the tube to provide substantially only the minimum clearance necessary to allow the disc to fall freely into the tube.
  • This minimum clearance will accommodate only a minimum amount of liquid reagent subsequently to be added to the tube, so that a highly concentrated dispersion of the cellular material may be formed in the tube immediately above the membrane-covered surface of the disc.
  • a typical luminometer cuvette has an internal diameter of the order of 9 mm whereas a conventional syringe may have a typical internal volume of up to 20 ml. Consequently, the concentration of the cellular material in the tube above the disc may be a factor of 20 or even 200 times greater than that in the syringe prior to the filtration.
  • the sample thus prepared is to be evaluated by means of a luminometer, the great majority of the dispersion formed in the tube will be above the disc which will consequently provide little interference to optical assay through the wall of the tube.
  • the disc assembled with its membrane, may be secured within the tubular body by a variety of means, provided that it may be released by remote manipulation into the collection vessel.
  • claws extend radially inwardly through apertures in the wall of the tubular body to urge the disc assembly axially onto the seating.
  • the claws may be manipulated externally of the tubular body to cause them to move radially outwardly to release the disc.
  • the disc assembly must close the passageway through the tubular body and the necessary seal may be provided by direct engagement of the peripheral portion of the membrane with a seating in the form of an annular shoulder in the body.
  • the tubular body may extend a sufficient distance downstream of the seating for the disc assembly to provide a guard which inhibits contact, and potential contamination, of the disc assembly.
  • This guard will necessarily have an internal cross-section large enough for the disc to fall through it into the collection vessel. However, its internal cross-sectional dimension may exceed the peripheral dimension of the disc sufficiently to accommodate the rim at the open end of a cuvette or other tubular connection vessel.
  • the tubular collection vessel will be inserted into the outlet end of the tubular body up to a position adjacent to the disc, before the disc is released, whereby the disc passes directly into the collection vessel.
  • Figure 1 is an exploded diagrammatic perspective view
  • Figure 2 is an enlarged view of part of Figure 1
  • Figure 3 is an axial cross-section through part of Figure 2 in greater detail than Figure 2.
  • the heart of the system is a device comprising a tubular body 4, having an inlet end 5 provided with a Luer-lock connector 6 and an outlet end 7.
  • the guard section 8 is formed with two diametrically opposed apertures 10.
  • the body 4 is a plastics moulding and is formed integrally with a pair of resilient claws 11 which extend radially inwardly through respective ones of the apertures 10, are connected to the tubular portion of the body 4 by respective bridges 12, and continue into respective fingerpieces 13 which overlie the outside of the body 4 with a clearance. The arrangement is such that the claws 11 can be moved radially outwardly through the apertures 10 by squeezing the fingerpieces 13 together.
  • a disc assembly D which, as shown in Figure 3 , consists of a skirt 14, a web 15 perforated by a number of holes 16, and circular wafer of a filter membrane 17 which is secured within a shallow recessed end of the disc.
  • One or more circular wafers 22 of loosely woven or other perforate compressible material may be interposed between the membrane wafer 17 and the base of the recess.
  • the claws 11 In the initial position in which the device is supplied, the claws 11 overlie the skirt 14 and hold the disc assembly on the seating 9 with the peripheral portion of membrane 17 in sealing engagement with the seating. Conformity of the membrane to the seating is improved by the compressibility of the wafer(s) of compressible material.
  • the radially outer peripheral surface 18 of the skirt 14 is cylindrical.
  • a syringe 19 having a neck 20 with a complementary Luer lock connector, is coupled to the inlet end of the body 4.
  • the syringe contains, for example, a primary filtered or other dispersion of cellular material in a liquid medium and when the syringe plunger is depressed, this is forced into the tubular body 4, and filtered by the membrane 17, the filtrate passing through the holes 16 and out through the outlet end 7 to waste.
  • the cellular material will simultaneously be retained by the membrane.
  • the operation will normally be carried out with the axis of the tubular body 4 substantially vertical with its inlet 5 uppermost.
  • the open end of a tubular collection vessel such as a luminometer cuvette 21 is inserted upwardly through the outlet end 7 of the body 4, within the guard 8 until it is adjacent to the claws 11.
  • a tubular collection vessel such as a luminometer cuvette 21
  • the guard 8 is stepped internally between a smaller diameter part 8A, which closely surrounds the disc D, and a larger diameter part 8B, which will accept the cuvette.
  • the finger pieces 13 are then squeezed towards one another, causing the claws 11 to be displaced radially outwardly, and releasing the disc assembly so that it falls into the cuvette 21, the cylindrical peripheral surface 18 of the disc skirt maintaining the disc at the same orientation within the cuvette, with the membrane facing upwardly.
  • An intensive concentrated dispersion of the cellular material for the appropriate analysis, such as bioluminescence, is then created by introducing into the cuvette just sufficient reagent to immerse the cellular material on the membrane, and to provide a sufficient depth of the dispersion within the cuvette above the membrane-covered surface of the disc.
  • the disc is a close fit within the cuvette so that there is very little wasted dispersion in the annular clearance between the periphery of the disc and the wall of the cuvette.
  • a typical aliquot of reagent for bioluminescence would comprise 100 ⁇ l of lysing agent followed by 100 ⁇ l of enzymatic agent.
  • the filter membrane 17 will be selected to have the appropriate pore size for the cellular material to be collected but may typically be a track-etched polycarbonate membrane, with a pore size in the order of 0.2 to 5 micron.
  • the system as illustrated provides the minimum of opportunity for the collected cellular material to be exposed to a contaminant, such as human-carried or environmental ATP.
  • a contaminant such as human-carried or environmental ATP.
  • the device consisting of a tubular body 4 with the disc assembly in place will be provided in a sterile and ATP-free packet.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Water Supply & Treatment (AREA)
  • General Health & Medical Sciences (AREA)
  • Pathology (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

L'invention se rapporte à un procédé et à un dispositif de concentration de matière cellulaire destiné à effectuer une analyse et comprenant un corps tubulaire contenant un disque poreux recouvert d'une membrane de filtration. On fait passer dans ce corps une dispersion de matière cellulaire de façon à ce que cette dernière s'amasse sur la membrane. Le disque auquel est fixée la membrane peut ensuite se détacher de façon à ce qu'il se dégage du corps tubulaire et tombe dans un réceptacle de récupération en vue d'analyser la substance cellulaire.
PCT/GB1996/001140 1995-05-23 1996-05-14 Ameliorations apportees a une technique de filtration WO1996037301A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9510395.8 1995-05-23
GBGB9510395.8A GB9510395D0 (en) 1995-05-23 1995-05-23 Improvements in filtration

Publications (1)

Publication Number Publication Date
WO1996037301A1 true WO1996037301A1 (fr) 1996-11-28

Family

ID=10774885

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1996/001140 WO1996037301A1 (fr) 1995-05-23 1996-05-14 Ameliorations apportees a une technique de filtration

Country Status (2)

Country Link
GB (1) GB9510395D0 (fr)
WO (1) WO1996037301A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2801660A1 (fr) * 1999-11-29 2001-06-01 Chemunex Dispositif de manipulation d'un porte-support solide pour contaminants
WO2003036267A2 (fr) * 2001-10-19 2003-05-01 Monogen, Inc. Procede et systeme de filtration destines a obtenir une couche cytologique
WO2008150779A1 (fr) * 2007-05-31 2008-12-11 3M Innovative Properties Company Dispositifs et procédés permettant de recueillir et de concentrer des échantillons en vue d'une analyse microbiologique
WO2009106760A2 (fr) * 2008-01-09 2009-09-03 Metagenex Dispositif et procédé pour isoler et cultiver des cellules vivantes sur filtre ou extraire leur matériel génétique
FR2935392A1 (fr) * 2008-09-02 2010-03-05 Metagenex Dispositif et procede pour isoler et cultiver des cellules vivantes sur filtre ou extraire leur materiel genetique
FR3012970A1 (fr) * 2013-11-12 2015-05-15 Biocarecell Dispositif de filtration destine a un liquide contenant des cellules en vue de les isoler.

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0438595A1 (fr) * 1989-07-12 1991-07-31 SIBASAKI, Michiro Bac et dispositif de filtrage
WO1992018844A1 (fr) * 1991-04-18 1992-10-29 La Mina Ltd. Recipient a enchantillons de liquide et ses modules de test
WO1994001213A1 (fr) * 1992-07-06 1994-01-20 Beckman Instruments, Inc. Colonne a reaction de synthese
WO1994005395A1 (fr) * 1992-08-27 1994-03-17 Gelman Sciences Inc. Ensemble cuve de filtration a membrane microporeuse

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0438595A1 (fr) * 1989-07-12 1991-07-31 SIBASAKI, Michiro Bac et dispositif de filtrage
WO1992018844A1 (fr) * 1991-04-18 1992-10-29 La Mina Ltd. Recipient a enchantillons de liquide et ses modules de test
WO1994001213A1 (fr) * 1992-07-06 1994-01-20 Beckman Instruments, Inc. Colonne a reaction de synthese
WO1994005395A1 (fr) * 1992-08-27 1994-03-17 Gelman Sciences Inc. Ensemble cuve de filtration a membrane microporeuse

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2801660A1 (fr) * 1999-11-29 2001-06-01 Chemunex Dispositif de manipulation d'un porte-support solide pour contaminants
WO2001040759A1 (fr) * 1999-11-29 2001-06-07 Chemunex Dispositif de manipulation de porteur de support solide pour contaminants
WO2003036267A2 (fr) * 2001-10-19 2003-05-01 Monogen, Inc. Procede et systeme de filtration destines a obtenir une couche cytologique
WO2003036267A3 (fr) * 2001-10-19 2003-09-04 Monogen Inc Procede et systeme de filtration destines a obtenir une couche cytologique
US7316779B2 (en) 2001-10-19 2008-01-08 Monogen, Inc. Filtration system and method for obtaining a cytology layer
WO2008150779A1 (fr) * 2007-05-31 2008-12-11 3M Innovative Properties Company Dispositifs et procédés permettant de recueillir et de concentrer des échantillons en vue d'une analyse microbiologique
WO2009106760A2 (fr) * 2008-01-09 2009-09-03 Metagenex Dispositif et procédé pour isoler et cultiver des cellules vivantes sur filtre ou extraire leur matériel génétique
WO2009106760A3 (fr) * 2008-01-09 2010-05-06 Metagenex Dispositif et procédé pour isoler et cultiver des cellules vivantes sur filtre ou extraire leur matériel génétique
JP2011509407A (ja) * 2008-01-09 2011-03-24 スクリーンセル 生きた細胞をフィルタ上で分離して培養するまたはその細胞の遺伝子材料を抽出するための装置と方法
US9339818B2 (en) 2008-01-09 2016-05-17 Screencell Device and method for isolating and cultivating live cells on a filter or extracting the genetic material thereof
FR2935392A1 (fr) * 2008-09-02 2010-03-05 Metagenex Dispositif et procede pour isoler et cultiver des cellules vivantes sur filtre ou extraire leur materiel genetique
FR3012970A1 (fr) * 2013-11-12 2015-05-15 Biocarecell Dispositif de filtration destine a un liquide contenant des cellules en vue de les isoler.

Also Published As

Publication number Publication date
GB9510395D0 (en) 1995-07-19

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