Apparatus for pretreating cells for flow cytometry

• US 5,167,926 A
• Filed: 09/11/1990
• Issued: 12/01/1992
• Est. Priority Date: 09/13/1989
• Status: Expired due to Fees

• Current Assignee: Kabushiki Kaisha Tiyoda Seisakusho
• Original Assignee: Kabushiki Kaisha Tiyoda Seisakusho
• Inventors: Kimura, Shiro, Miyazawa, Kazuo
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): Soderquist, Arlen
• Application Number:
  US07/580,864
• Time in Patent Office: 812 Days
• Field of Search: 422/64, 422/67, 422/68.1, 422/72, 422/81, 422/100, 422/101, 436/45, 436/177
• US Class Current: 422/67
• CPC Class Codes: G01N 1/28 Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 2035/00376 Conductive heating, e.g. heated plates
• CPC Class Codes: G01N 2035/00435 Refrigerated reagent storage
• CPC Class Codes: G01N 2035/00504 combined with carousels
• CPC Class Codes: G01N 2035/0441 for samples
• CPC Class Codes: G01N 2035/1053 for separating part of the liquid, e.g. filters, extraction phase
• CPC Class Codes: G01N 35/02 using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations G01N35/0098 and G01N35/0099 take precedence
• CPC Class Codes: Y10T 436/111666 Utilizing a centrifuge or compartmented rotor
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

A plurality of open-top sample tubes, each containing a simple liquid containing cells to be treated, are removably mounted on a disklike sample carrier which is rotatable about a vertical axis for centrifuging the samples. Disposed above the sample carrier are a set of fixed reagent supply nozzles for dropping a desired reagent or reagents into the sample tubes and at least one supply-discharge nozzle movable into and out of the successive sample tubes. A hydropneumatic circuit is coupled to the supply-discharge nozzle for intimately intermingling the samples and the reagents and for rinsing the supply-discharge nozzle and other required parts. In an alternate embodiment a single supply-discharge nozzle is employed for reagent supply into the sample tubes, for intermingling the samples and the reagents, and for rinsing.

Claims

1. An apparatus for treating cells preparatory to a cytological study, said apparatus comprising:

(a) sample containers for containing a sample liquid containing cells to be treated;

(b) reagent containers for containing one or more reagents to be added to the sample liquid;

(c) a rinsing liquid container for container a rinsing liquid;

(d) pneumatic energy source means for developing pneumatic energy;

(e) a horizontal rotary sample carrier of a substantially disk shape rotatable about a vertical axis,

(f) a sample rack of a substantially arcuate elongated shape mounted on said sample carrier along a peripheral edge thereof, said sample rack being pivoted at two ends thereof to the sample carrier so as to be swingable radial outwardly of the sample carrier under centrifugal force when the carrier is rotated about said vertical axis, said sample rack having therealong a row of holes for receiving said sample containers;

(g) drive means for imparting rotation about said vertical axis to the sample carrier to centrifuge the sample containers received in the holes of said sample rack;

(h) constant temperature vessels of substantially arcuate elongated shape, said vessels having thermostatic means for maintaining a constant desired temperature and being provided horizontally under said sample carrier, said vessels being disposed in series in annular arrangement below a peripheral edge of the sample carrier, each of said vessels being movable vertically relative to the sample carrier to a position in which the sample containers carried by the sample rack are inserted into one of the vessels for application of a desired temperature to the sample containers;

(i) a row of reagent supply nozzles disposed above the sample carrier along a peripheral edge thereof for simultaneously dropping the reagents into the sample containers on the sample rack to form sample-reagent mixtures in the sample containers;

(j) a selector valve for selectively supplying the reagents and the rinsing liquid from said reagent containers and said rinsing liquid container into the sample containers;

(k) a supply-discharge nozzle capable of communication with said selector valve and the pneumatic energy source means and movable into and out of the sample containers for introducing any of the one or more reagents or the rinsing liquid into the sample containers, and for withdrawing the sample-reagent mixtures or the rinsing liquid from the sample containers, under the force of the pneumatic energy;

(l) positioning means coupled to said sample carrier for angularly indexing the carrier about the vertical axis thereof so as to cause the sample containers on the sample rack to take positions in vertical alignment with said reagent supply nozzles, said supply-discharge nozzle and said constant-temperature vessels;

(m) a temporary storage vessel capable of communication with the supply-discharge nozzle; and

(n) valve means for controlling the communication of the supply-discharge nozzle with the selector valve and the pneumatic energy source means and the temporary storage vessel to intimately intermingle the sample liquid and the reagents by causing the flow of the sample-reagent mixtures back and forth between the sample containers and the temporary storage vessel, and to rinse the nozzle means and the temporary storage with the rinsing liquid.

2. The cell treating apparatus of claim 1, wherein each of the sample containers has an open top, and wherein the supply-discharge nozzle has a flange formed thereon for hermetically closing the open top of one of the sample containers when fully inserted therein.

3. The cell treating apparatus of claim 1, wherein the pneumatic energy source means comprises means for developing both positive and negative pneumatic pressures.

4. The cell treating apparatus of claim 3, wherein the positive pneumatic pressure is greater in absolute value than the negative pneumatic pressure.

5. The cell treating apparatus of claim 1, further comprising:

(a) a first sample filter disposed between the temporary storage vessel and the supply-discharge nozzle for breaking up coherent cell aggregates into individual cells; and

(b) a second sample filter of finger mesh than the first sample filter, the second sample filter being disposed between the temporary storage vessel and the pneumatic energy source means for arresting the individual cells.