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WO1993011419A1 - Procede et dispositif de preparation d'echantillons de tissus et de cellules - Google Patents

Procede et dispositif de preparation d'echantillons de tissus et de cellules Download PDF

Info

Publication number
WO1993011419A1
WO1993011419A1 PCT/SE1992/000813 SE9200813W WO9311419A1 WO 1993011419 A1 WO1993011419 A1 WO 1993011419A1 SE 9200813 W SE9200813 W SE 9200813W WO 9311419 A1 WO9311419 A1 WO 9311419A1
Authority
WO
WIPO (PCT)
Prior art keywords
slides
holder
samples
tissue samples
staircase
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/SE1992/000813
Other languages
English (en)
Inventor
Christer Busch
Jan SÄLLSTRÖM
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
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 Individual filed Critical Individual
Publication of WO1993011419A1 publication Critical patent/WO1993011419A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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
    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31—Apparatus therefor
    • G01N1/312—Apparatus therefor for samples mounted on planar substrates

Definitions

  • the present invention relates to a method and a device for preparing cell and tissue samples for subsequent analysis.
  • Histological samples are prepared by embedding tissue samples in paraffin, cutting in thin sections and, after fixating, arranging on slides. Alternatively, sections on slides can be made from frozen tissue. Thereafter, the sample is prepared for analysis which starts with removing the paraffin and converting the sample to an aqueous phase. This preparation, or 'staining', continues with incubating of the samples with different reagents in a specific order prior to final analysis.
  • Some types of staining methods such as immunohistochemistry and in situ hybridization, the purpose of which is to specifically stain single biochemical components in the preparation, are performed by a large number of different preparation steps and with small volumes of expensive reagents.
  • slide holding devices are known in which the slides are arranged in pairs with the sample provided sides of each slide in a pair facing each other to form a chamber being filled with reagents by capillary action.
  • the drawback with this approach is that it requires relatively large volumes and high concentrations of expensive reagents.
  • the slides are carefully put back into the holder one by one to avoid the risk of mixing up the slides.
  • the next step is usually a PBS wash.
  • the slides are taken out of the holder, carefully as above, and the samples provided with biotinylated antibody being pipetted on the sample.
  • the slides are again taken out of the holder, this time to be provided with small amounts of ABC (avidin, biotin) complex, and then inserted into the holder for a repeated PBS wash.
  • ABC avidin, biotin
  • the enzyme development occurs in a closed hood.
  • the slides are taken out of the holders to provide the samples with an enzyme substrate, thereafter into the holders again for PBS wash.
  • the slides have to be removed from and inserted into the holders in a total of three to four times. Besides the time and labour demanding aspects of this, there is also the risk of mixing ,up the slides.
  • Fig. 1 is a schematic view of a device according to the in ⁇ vention
  • Fig. 2 is a section view along line II-II in Fig. 1 of the holder shown in Fig 1;
  • Fig. 3 is a schematic view of a preparation system comprising the device according to Fig. 1.
  • a device comprising a holder 2 with inserted slides 3 each provided with sectioned samples on the upper side.
  • the samples are arranged at the same location on every slide, usually close to one of the shorter side edges.
  • the holder 2 is open at both ends and the slides 3 extend a distance through both ends of the holder.
  • the device 1 also comprises a staircase formation unit 4 having, for example, six steps as shown.
  • the holder is gripped and the slides are pressed against the steps of the unit 4 in the- direction of the arrow in Fig. 1 to obtain the shown staircase form.
  • This form is used for pipetting small volumes of reagents on each sample individually.
  • Fig. 2 shows a section view of the holder 2.
  • the slides 3 are securely held by resilience of tongues 5 forming grooves into which the slides 3 are inserted.
  • a system for preparing tissue or cell samples is shown.
  • the holder 2 and unit 4 are releasably arranged on a bottom plate 6.
  • the holder 1 can optionally be connected with one or more other holders.
  • the protrusions 8 on the holder 9 are inserted into suitably dimensioned holes of another holder for connection.
  • These holders are then placed cuvettes 9 being dimensioned for more than one cuvette.
  • An incubation chamber 10 is provided to cover the holder 2 and slides 3 during incubation. To absorbe any moist on the slides following incubation, there is provided an absorbtion chamber 11 with absorbating material in the bottom.
  • the positions on plate 6 can be varied enabling mounting of an optional reaction and wash chain.
  • the slides 3 are alternately arranged in a straight form, ie. uniform length of each slide protrudes out of the holder, and in the staircase form according to Fig. 1, ie. different lengths of each slide is protruding out of the holder, the straight form being used for dipping of the slides into different baths and staircase form, in which the holder is rotated 90° in relation to the straight form, is used to apply small amounts of reagents to the samples on the slides.
  • the device 1 can be used either in a manual or in an automatic system.
  • the present invention does not require removing of the slides from the holder at any time during the preparation steps and, thus, the mix up risk is avoided.
  • the staircase form enables access to every individual slide sample. Having completed the addition of small amounts of ragents, the slides are again arranged to the initial position, in which the slides are arranged in a straight form. Because the insertion and removal steps into and out of the holders are eliminated, the method furthermore becomes time and labour saving and requires a minimum of reagents.
  • the holder and its tongues are preferably made of plastic with deformation zones or rubber or other materials with ability to take up different tolerances of the slides or other sample supports.
  • the device and method according to the invention are suited for all types of stainings, for example immunocytochemical staining, enzyme histochemical staining and in situ hybridization.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

La présente invention se rapporte à un procédé de préparation d'échantillons de cellules ou de tissus destinés à une analyse, lequel consiste à appliquer lesdits échantillons sur des lames ou d'autres supports, à fixer lesdites lames dans un dispositif de retenue, et à soumettre les échantillons à des volumes relativement importants de solutions tampons et à de faibles volumes de solutions réactives. Le procédé se caractérise en ce que les lames sont disposées en atlernance de manière verticalement alignée et en forme d'escalier, forme dans laquelle différentes longueurs de chaque lame font saillie hors du dispositif de retenue pour permettre d'obtenir un accès individuel auxdits échantillons. L'invention se rapporte également à un dispositif et à un système destinés à être utilisés selon ce procédé.
PCT/SE1992/000813 1991-11-25 1992-11-25 Procede et dispositif de preparation d'echantillons de tissus et de cellules Ceased WO1993011419A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE9103483A SE469148B (sv) 1991-11-25 1991-11-25 Foerfarande och anordning foer att preparera cell- eller vaevnadsprov i analyssyfte
SE9103483-5 1991-11-25

Publications (1)

Publication Number Publication Date
WO1993011419A1 true WO1993011419A1 (fr) 1993-06-10

Family

ID=20384421

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE1992/000813 Ceased WO1993011419A1 (fr) 1991-11-25 1992-11-25 Procede et dispositif de preparation d'echantillons de tissus et de cellules

Country Status (2)

Country Link
SE (1) SE469148B (fr)
WO (1) WO1993011419A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2058128A (en) * 1934-10-08 1936-10-20 Howard F Brubach Slide holder
GB2119119A (en) * 1982-04-28 1983-11-09 Frank W Jackson Apparatus and methods of staining biological slides
EP0293076A2 (fr) * 1987-04-30 1988-11-30 ERIE SCIENTIFIC COMPANY (a Delaware Corporation) Réceptacle pour plaques de verre minces
EP0414405A2 (fr) * 1989-08-21 1991-02-27 Fisher Scientific Company Support de porte-objets alignés et assemblage de porte-objets

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2058128A (en) * 1934-10-08 1936-10-20 Howard F Brubach Slide holder
GB2119119A (en) * 1982-04-28 1983-11-09 Frank W Jackson Apparatus and methods of staining biological slides
EP0293076A2 (fr) * 1987-04-30 1988-11-30 ERIE SCIENTIFIC COMPANY (a Delaware Corporation) Réceptacle pour plaques de verre minces
EP0414405A2 (fr) * 1989-08-21 1991-02-27 Fisher Scientific Company Support de porte-objets alignés et assemblage de porte-objets

Also Published As

Publication number Publication date
SE9103483L (sv) 1993-05-17
SE9103483D0 (sv) 1991-11-25
SE469148B (sv) 1993-05-17

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