Cell selecting apparatus

• US 5,106,584 A
• Filed: 11/16/1989
• Issued: 04/21/1992
• Est. Priority Date: 09/18/1984
• Status: Expired due to Fees

• Current Assignee: Sumitomo Electric Industries Limited (Sumitomo Corporation)
• Original Assignee: Sumitomo Electric Industries Limited (Sumitomo Corporation)
• Inventors: Funakubo, Hiroyasu, Nishiwaki, Yoshikazu, Miyake, Shinichi
• Primary Examiner(s): Lacey, David L.
• Assistant Examiner(s): Trautman, Kimberly A.
• Application Number:
  US07/437,287
• Time in Patent Office: 887 Days
• Field of Search: 422/63, 422/64, 422/65, 422/67, 422/73, 436/47, 436/63, 436/69, 356/39, 356/72, 356/73, 356/335, 356/336, 356/337, 356/338, 356/339, 356/340
• US Class Current: 422/65
• CPC Class Codes: C12M 23/04 Flat or tray type, drawers C12M23/10, C12M23/12, C12M23/16 take precedence
• CPC Class Codes: C12M 23/50 Means for positioning or orientating the apparatus C12M41/08 takes precedence
• CPC Class Codes: C12M 33/04 by injection or suction, e.g. using pipettes, syringes, needles pipettes in general B01L3/02
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: G01N 35/028 having reaction cells in the form of microtitration plates
• CPC Class Codes: Y10T 436/113332 with conveyance of sample along a test line in a container or rack

Abstract

A cell selecting apparatus which has an extracting and injection section, a tray transport section, a cultivating section, a cell selecting section, and a controller for controlling each section according to the operating conditions which are inputted therein. This arrangement makes it possible to select and isolate the desired cells or microorganisms from mixtures of different types of cells or microorganisms, for example, those which are capable of proliferating over short periods of time and producing antibodies or secretions, from a liquid culture of cells.

Claims

1. A cell selecting apparatus, comprising:

an extracting and injecting section;

a tray transport section for transporting at least one tray having a plurality of wells;

a controller; and

a

cell selecting section,

wherein said extracting and injecting section comprises a pipette, a manipulator for holding and moving said pipette up and down at a predetermined position above a culture tray and a pump for drawing and discharging a predetermined quantity of a liquid culture into and from said pipette,

said tray transport section comprises a transport section tray support for the support thereon of the culture tray provided with a plurality of wells therein, a first tray chuck and a first drive means for transporting said transport section tray support to predetermined positions with respect to said extracting and injecting section,

said cell selecting section comprises a cultivating section and a colony observatory section, wherein said cultivating section comprises an incubator including a tray stock for accommodating a plurality of trays, a second tray chuck and a second drive means for moving said stock and a cultivating section tray support for transporting cultivating trays to said tray stock, and further said cultivating section being operable to interact with said extracting and injecting section in a manner such that said pipette can be manipulated to a predetermined position at which location said liquid culture may be injected into or extracted from a cell colony without disturbing the growth of said cell colony, and

said controller comprises a device which generates and transmits at least eight control signals for controlling each of said sections according to operating conditions of each of said sections inputted thereto, whereby said sections of said cell selecting apparatus operably interrelate and communicate sequentially in steps including,

(a) in response to a first signal and a second signal, moving said pipette to a position with respect to a container of cell solution so that, upon activation, said pump extracts a quantity of said cell solution from the container via said pipette,

(b) in further response to said second signal, moving said pipette to a position above a well of a designated culture tray,

(c) in response to a third signal, activating said pump to inject said cell solution contained in said pipette into a well of the designated culture tray,

(d) in response to a fourth signal, moving the designated culture tray a distance corresponding to one of said wells,

(e) in response to said fourth signal and a fifth signal, sixth signal and seventh signal, transferring the designated culture tray to a position within said incubator of said cultivating section,

(f) in response to an eight signal, providing a growth environment within said cultivating section as designated by parameter input to said controller, and

(g) in further response to said fourth, fifth, sixth and seventh signals, moving the designated culture tray to said colony observatory section, and in response to a first measurement signal from said colony observatory section, calculating as a value, the number and size of said cell colonies which have been processed.

2. The cell selecting apparatus as defined in claim 1 wherein said colony observatory section comprises:

a TV carmera, and

a control circuit for converting into an electric signal a surface area occupied by colonies whose image has been received by said TV camera.

3. A cell selecting apparatus, comprising:

a tray transport section for transporting at least one tray;

an extracting and injecting section;

a cell selecting section; and

a controller,

wherein said tray transport section comprises a transport section tray support for supporting thereon a culture tray provided with a plurality of wells therein, a tray chuck and a first drive means for transporting said tray support to predetermined positions with respect to said extracting and injecting section and said cell selecting section,

said extracting and injecting section comprises a pipette, a manipulator for holding and moving said pipette up and down with respect to a predetermined position above said culture tray and a pump for drawing and discharging a predetermined quantity of a cell fluid into and from said pipette,

said cell selecting section comprises a colony observatory section and an assay section, said assay section comprising and antibody detecting tray support for supporting thereon an antibody detecting tray provided with a plurality of walls therein, and a second drive means for grasping and transporting said antibody detecting tray onto said antibody detecting tray support, and a detector selected from group consisting of means for detecting the degree of color development by an enzyme antibody method or a means for defining spectra of liquid chromatography; and

said controller comprises a device which generates and transmits at least five control signals for controlling each of said sections according to operating conditions of each of said sections inputted thereto, whereby said sections of said cell selecting apparatus operably interrelate and communicate in sequential steps including,

(a) in response to a second signal, positioning said culture tray on said transport section tray support by said tray chuck,

(b) in response to a first signal, moving said culture tray by said first drive means to said colony observatory section, and in response to a first measurement signal from said colony observatory section, measuring number and size of colonies,

(c) in response to a third signal, moving said pipette to a position above a well provided in said culture tray,

(d) in response to a fourth signal, extracting a cell supernatant from said well in said culture tray via said pump,

(e) in further response to said third signal, moving said pipette containing said cell supernatant to a position above a well of said antibody detecting tray,

(f) in further response to said fourth signal, injecting said cell supernatant from said pipette into said well of said antibody detecting tray,

(g) in response to said first and second signals and a fifth signal, transporting said antibody detecting tray to said assay section by said second drive means, and

(h) in response to a second measurement signal from said assay section, measuring the quantity of antibodies contained in said cell supernatant and storing this information in said controller, and calculating the antibody producing capability of the cells from this value.

4. The cell selecting apparatus as defined in claim 3 wherein said detector comprises:

an ELISA analyzer.

5. A cell selecting apparatus, comprising:

an extracting and injecting section;

a tray transport section for transporting at least one tray having a plurality of wells;

a cell selecting section; and

a controller,

wherein said extracting and injecting section comprises a pipette, a manipulator for holding and moving said pipette up and down at a predetermined position above a cell culture tray and a pump for drawing and discharging a predetermined quantity of a cell solution into and from said pipette,

said tray transport section comprises a tray support for supporting thereon a cell culture tray provided with a plurality of wells, a tray chuck and a first drive means for transporting said tray support to predetermined positions with respect to said extracting and injecting section and cell selecting section,

said cell selection section comprises a cell number counting section and a cell isolating section, said cell number counting section comprises a glass plate transport section and a second drive means for moving said glass plate from a position below said pipette at which said cell solution is dropped, to an observatory position for a microscope where said cell solution on said glass plate is observed, and a control circuit for counting the number of cells observed by the microscope through a TV camera,

said cell isolating section comprises a diluting vessel into which said cell solution extracted by said extracting and injecting section from wells of said cell culture tray having a predetermined number of cells specified by said cell number counting section through said controller is injected, and said cell isolating and said injecting section having a cell solution passage into which diluted cell solution is drawn from said diluting vessel, and a cell detecting section disposed in said passage, said tray transport section is operable in synchronism with a cell detection signal from a cell detecting section to move said cell culture tray a distance corresponding to the space between said wells at an outlet of said cell solution passage, and

said controller comprises a device which generates and transmits at least six controlling each said section according to operating conditions for each said section inputted thereto, whereby said sections of said cell selecting apparatus operably interrelate and communicate in sequential steps including,

(a) in response to a first signal, moving said pipette to a position above one of said wells of said cell culture tray via said manipulator of said extracting and injecting section,

(b) in response to a second signal, extracting said cell solution from said one of said wells of said cell culture tray,

(c) in further response to said first signal, positioning said pipette containing said cell solution above said glass plate of said cell number counting section,

(d) in further response to said second signal, injecting said cell solution contained in said pipette onto said glass plate of said cell number counting section,

(e) in response to a fifth signal, moving said glass plate to a position below the microscope, and in response to a first measurement signal from said cell number counting section, counting the number of cells viewable within a field of sight on said glass plate and calculating a concentration of cells in said cell solution,

(f) in further response to said first signal, returning said pipette to said cell culture tray,

(g) in further response to said second signal, extracting said cell solution from one of said wells of said cell culture tray,

(h) in further response to said first signal, moving said pipette to said diluting vessel via said manipulator of said extracting and injecting section,

(i) in further response to said second signal, injecting said cell solution into said diluting vessel,

(j) in response to a sixth signal, adding diluting liquid to said diluting vessel, and in response to a second measurement signal, detecting the passage of cells in said diluted cell solution past a detecting position,

(k) in further response to said third and fourth signals, moving a second culture tray of said tray transport system a distance of one of said wells of said cell culture tray at the outlet of said cell solution passage.

6. The cell selecting apparatus as defined in claim 5 wherein said cell detecting section comprises:

a pair of electrodes on respective sides of said cell solution passage.

7. The cell selecting apparatus as defined in claim 5 wherein said cell detecting section comprises:

a light emitter and a light receiver positioned on respective sides of said cell solution passage.

8. A cell selecting apparatus, comprising:

an extracting and injecting section;

a tray transport section for transporting at least one cell culture tray;

a controller; and

a cell selecting section,

wherein said extracting and injecting section comprises a pipette, manipulator for holding the pipette and for moving the pipette up and down at a predetermined position above said culture tray and a pump for drawing and discharging a predetermined quantity of a cell solution into and from said pipette,

said tray transport section comprises a transport section tray support for supporting thereon one said cell culture tray provided with a plurality of wells therein, a first tray chuck and a first drive means for transporting one said cell culture tray to predetermined positions with respect to said extracting and injecting section and said cell selecting section,

said cell selecting section comprises a cultivating section, a colony observation section, an assay section, a cell number counting section and a cell isolating section,

wherein said cultivating section comprises an incubator including a tray stock for accommodating a pluralilty of said cell culture trays, a second tray chuck and a second drive means for moving said tray stock and a cultivating section tray support for transporting said cell culture trays to said tray stock, and further said cultivating section being operable to interact with said extracting and injecting section in a manner such that said pipette can be manipulated to a predetermined position at which location said cell solution may be injected into or extracted from a cell colony without disturbing the growth of said colony,

said assay section comprises an antibody detecting tray support for supporting thereon an antibody detecting tray provided with a plurality of wells therein, and said second drive means for grasping and transporting said antibody detecting tray onto said antibody detecting tray support, and a detector selected from the group consisting of means for detecting the degree of color development by an enzyme antibody method or a means for defining spectra of liquid chromatography,

said cell number counting section comprises a glass plate transport section and a third drive means for moving said glass plate from a position below said pipette at which said cell solution is dropped, to an observatory position for a microscope where said cell solution on said glass plate is observed, and a control circuit for counting the number of cells observed by the microscope through a TV camera,

said cell isolating section comprises a diluting vessel into which said cell solution extracted by said extracting and injecting section from wells of said cell culture tray in which said cells secrete antibodies, and said cell isolating section having a cell solution passage into which diluted cell solution is drawn from said diluting vessel and a cell detecting section disposed in said passage, and said tray transport section is operable in synchronism with a cell detecting signal from said cell detecting section to move said cell culture tray a distance corresponding to the space between said wells at an outlet of said cell solution passage,

and said controller comprises a device which generates and transmits control signals for controlling each said section according to operating conditions for each said inputted thereto, whereby said section of said cell selecting apparatus operably interrelate and communicate in sequential steps including,

(a) in response to a first measurement from said colony observatory section, measuring number and size of colonies, and replacing the supernatant of the cell culture tray to fresh liquid culture or drawing said supernatant of said cell solution to detect the quantity of antibody,

(b) in response to a second measurement signal from said assay section, measuring the quantity of antibody of said wells of the cell culture tray and detecting the well producing the greater quantity of antibody, and drawing the cell solution of the well to measure the cell number of the cell solution,

(c) in response to a third measurement signal from said cell number counting section, measuring the cell number and determining the degree of dilution with respect to a predetermined concentration, and diluting the cell solution to isolate the cells.