KR920703785A - Automated testing equipment - Google Patents

Automated testing equipment

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Publication number
KR920703785A
KR920703785A KR1019920700118A KR920700118A KR920703785A KR 920703785 A KR920703785 A KR 920703785A KR 1019920700118 A KR1019920700118 A KR 1019920700118A KR 920700118 A KR920700118 A KR 920700118A KR 920703785 A KR920703785 A KR 920703785A
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KR
South Korea
Prior art keywords
incubator
mixture
cell
cells
delivering
Prior art date
Application number
KR1019920700118A
Other languages
Korean (ko)
Inventor
필러 크리스티안
이. 레히트 프라이드 프란쯔
Original Assignee
존 알. 스텝헨손
온코젠 사이언스 인크
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Application filed by 존 알. 스텝헨손, 온코젠 사이언스 인크 filed Critical 존 알. 스텝헨손
Publication of KR920703785A publication Critical patent/KR920703785A/en

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    • 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
    • C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/12—Well or multiwell plates
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/028—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations having reaction cells in the form of microtitration plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00178—Special arrangements of analysers
    • G01N2035/00207—Handling bulk quantities of analyte
    • G01N2035/00217—Handling bulk quantities of analyte involving measurement of weight
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00346—Heating or cooling arrangements
    • G01N2035/00455—Controlling humidity in analyser
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/00584—Control arrangements for automatic analysers
    • G01N35/00722—Communications; Identification
    • G01N35/00732—Identification of carriers, materials or components in automatic analysers
    • G01N2035/00742—Type of codes
    • G01N2035/00752—Type of codes bar codes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices
    • G01N2035/1032—Dilution or aliquotting
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices
    • G01N2035/1048—General features of the devices using the transfer device for another function
    • G01N2035/1058—General features of the devices using the transfer device for another function for mixing
    • G01N2035/106—General features of the devices using the transfer device for another function for mixing by sucking and blowing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0099—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor comprising robots or similar manipulators

Abstract

내용 없음No content

Description

자동화 실험장치Automated testing equipment

[도면의 간단한 설명][Brief Description of Drawings]

제1도는 본 발명에 따라 구성된 장치의 전체 배치도를 도시한 평면도,1 is a plan view showing an overall layout of a device constructed in accordance with the invention,

제2도는 제1도의 일부를 형성하며, 본 발명에 따라 사용된 액체 조작부의 사시도,Figure 2 forms part of Figure 1, a perspective view of the liquid operation section used in accordance with the present invention,

제3도는 본 발명의 전체적인 공정도를 나타낸 개략적인 플루우챠트,Figure 3 is a schematic flue chart showing the overall process diagram of the present invention,

제4도는 제1도의 장치에 사용되며, 기판과 주기판상에 포함된 32개의 표본약병 래크를 도시한 평면도.4 is a plan view showing 32 sample vial racks used on the apparatus of FIG. 1 and contained on a substrate and a main substrate.

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is an open matter, no full text was included.

Claims (16)

내부공간을 실제로 둘러싸는 하우징 수단으로 형성되고, 최소한 하나이상의 생체세포의 수집물을 생산 및 재생시키기에 적합한 규정된 조건을 상기 내부공간내에 유지하는 인큐베이터 수단과; 혼합물을 형성하기 위하여 화학약품과 세포를 혼합하기 위한 혼합수단으로 이루어진 조작수단과; 배양후 상기 혼합물을 수용하고 감지 가능한 변화에 응답하여 감지신호를 발생하는 감지수단과; 상기 인큐베이터수단에서, 상기 조작수단으로 세포를 전달하고, 규정된 조건하에 저장된 혼합물을 형성하도록 상기 세포와 혼합될 경우 상기 세포에서 감지가능한 변화를 발생하는 화학약품을 전달하며, 배양을 위하여 인큐베이터 수단내에서 상기 혼합물을 상기 인큐베이터 수단으로 전달하는 로보트 수단; 으로 구성된 것을 특징으로 하는 자동화 실험장치.Incubator means formed by housing means that actually enclose the interior space and maintaining defined conditions within the interior space suitable for producing and reproducing at least one collection of biological cells; Operating means comprising mixing means for mixing the chemical and the cells to form a mixture; Sensing means for receiving the mixture after incubation and generating a detection signal in response to a detectable change; In the incubator means, delivering the cells to the manipulating means, delivering a chemical that produces a detectable change in the cells when mixed with the cells to form a stored mixture under defined conditions, and in the incubator means for culturing Robot means for delivering said mixture to said incubator means; Automated experimental apparatus, characterized in that consisting of. 내부공간을 실제로 둘러싸는 하우징 수단으로 형성되고, 최소한 하나이상의 생체세포의 수집물을 생산 및 재생시키기에 적합한 규정된 조건을 상기 내부공간내에 유지하는 인큐베이터 수단과 규정된 조건하에 저장된 혼합물을 형성하도록 상기 세포와 혼합될 경우 상기 세포에서 감지가능한 변화를 발생하는 화학약품과 세포를 상기 인큐베이터 수단내에 수용할 수 있으며, 상기 인큐베이터 수단내에서 배양을 위하여 그 수단으로 전달된 혼합물을 형성하도록 상기 화학약품과 상기 세포를 혼합하기 위한 혼합수단으로 이루어진 조작수단과; 감지가능한 변화에 응답하여 감지신호를 발생하는 감비수단과; 상기 인큐베이터 수단내에서 배양후 상기 혼합물을 상기 감지수단으로 전달하는 로보트 수단; 으로 구성된 것을 특징으로 하는 자동화 실험장치.Formed to form a housing means that actually encloses the interior space and to form a mixture stored under the specified conditions with an incubator means that maintains within the interior space a defined condition suitable for producing and reproducing at least one collection of biological cells. The chemical and the cell may be contained in the incubator means when mixed with the cell to produce a detectable change in the cell and to form a mixture delivered to that means for culturing in the incubator means. Operating means comprising mixing means for mixing cells; Subtracting means for generating a sensing signal in response to the detectable change; Robot means for delivering the mixture to the sensing means after incubation in the incubator means; Automated experimental apparatus, characterized in that consisting of. 내부공간을 실제로 둘러싸는 하우징 수단으로 형성되고, 최소한 하나이상의 생체세포 수집물을 생산 및 재생시키기에 적합한 규정된 조건을 상기 내부공간내에 유지하는 인큐베이터 수단과; 혼합물을 형성하기 위하여 화학약품과 세포를 혼합하기 위한 혼합수단으로 이루어진 조작수단과; 감지가능한 변화에 응답하여 감지신호를 발생하는 감지수단과; 상기 인큐베이터수단에서 상기 조작수단으로 세포를 전달하고, 규정된 조건하에 저장된 혼합물을 형성하도록 상기 세포와 혼합될 경우 상기 세포에서 감지가능한 변화를 발생하는 화학약품을 전달하며, 인큐베이터 수단내에서 배양을 위하여 상기 인큐베이터 수단으로 전달하고, 배양후 상기 혼합을 상기 감지수단으로 전달하는 로보트 수단; 으로 구성된 것을 특징으로 하는 자동화 실험장치.Incubator means formed by housing means that actually enclose the interior space and maintaining defined conditions within the interior space suitable for producing and regenerating at least one biological cell collection; Operating means comprising mixing means for mixing the chemical and the cells to form a mixture; Sensing means for generating a sense signal in response to a detectable change; Delivering the cells from the incubator means to the manipulating means, delivering a chemical which, when mixed with the cells to form a stored mixture under defined conditions, produces a detectable change in the cells, for culturing in the incubator means Robot means for delivering to the incubator means and delivering the mixture to the sensing means after incubation; Automated experimental apparatus, characterized in that consisting of. 제1항 내지 제3항 중 어느 한 항에 있어서, 상기 인큐베이터 수단은 내부공간을 실온 37℃로 유지하기 위한 온도 제어수단으로 갖는 것을 특징으로 하는 자동화 실험장치.The automated experimental apparatus according to any one of claims 1 to 3, wherein the incubator means has a temperature control means for maintaining an internal space at room temperature of 37 ° C. 제4항에 있어서, 상기 온도제어수단은 상기 인큐베이터 수단의 일부에 가열하기 위한 적외선 가열수단을 갖도록 된 것을 특징으로 하는 자동화 실험장치.5. The automated experimental apparatus according to claim 4, wherein the temperature control means has infrared heating means for heating a part of the incubator means. 제1항 내지 제3항중 어느 한 항에 있어서, 상기 인큐베이터 수단은 내부공간을 상대 습도 약 100%로 유지하기 위한 습도제어수단을 갖도록 된 것을 특징으로 하는 자동화 실험장치.The automated laboratory apparatus according to any one of claims 1 to 3, wherein the incubator means has humidity control means for maintaining the internal space at about 100% relative humidity. 제6항에 있어서, 상기 습도제어수단은 입구 및 출구가 형성되어 상기 입구를 통하여 습한공기를 상기 내부공간내로 불어들임과 동시에 상기 출구를 통하여 상기 내부공간에서 불어내기 위하여 상기 내부공간과 연통되며, 상기 입구 및 출구는 서로 동축상으로 배치되는 커플링 수단을 갖도록 된 것을 특징으로 하는 자동화 실험장치.The method of claim 6, wherein the humidity control means is formed in the inlet and outlet through the inlet to blow the humid air into the inner space and at the same time communicate with the inner space through the outlet to blow out of the inner space, And said inlet and outlet have coupling means arranged coaxially with each other. 제1항 내지 제3항중 어느 한 항에 있어서, 상기 로보트 수단은 한정된 작업공간을 가지고, 상기 하우징수단은 다수개의 칸막이로 형성되며, 상기 인큐베이터 수단은 상기 하우징수단에 회전가능하게 설치되어 각기 다른 칸막이들이 상기 작업공간내로 선택적으로 이동되도록 하는 회전테이블 수단을 갖도록 이루어짐을 특징으로 하는 자동화 실험장치.According to any one of claims 1 to 3, wherein the robot means has a limited working space, the housing means is formed of a plurality of partitions, the incubator means are rotatably installed in the housing means different partitions Automated experimental apparatus, characterized in that it has a rotary table means for selectively moving into the workspace. 제1항 내지 제3항중 어느 한 항에 있어서, 상기 세포는 별도의 세포웰내에 넣어지고, 상기 혼합수단은 상기 각종 화학약품 희석액을 준비하고 상기 각종 희석액을 각각 다른 상기 세포웰에 첨가하는 노즐수단을 갖도록 함을 특징으로 하는 자동화 실험장치.The nozzle means according to any one of claims 1 to 3, wherein the cells are placed in separate cell wells, and the mixing means prepares the various chemical dilutions and adds the various dilutions to the different cell wells. Automated experiment apparatus characterized in that to have. 제9항에 있어서, 상기 노즐수단은 화학약품을 각각 투약하고 기판웰내로 희석시키기 위한 다수개의 제1노즐을 갖도록 됨을 특징으로 하는 자동화 실험장치.10. The automated laboratory apparatus according to claim 9, wherein the nozzle means has a plurality of first nozzles for respectively dosing chemicals and diluting them into a substrate well. 제10항에 있어서, 상기 노즐수단은 상기 희석액을 상기 세포웰로 투약하기 위한 다수개의 제2노즐을 갖도록 됨을 특징으로 하는 자동화 실험장치.11. The automated laboratory apparatus according to claim 10, wherein the nozzle means has a plurality of second nozzles for administering the diluent to the cell well. 제10항에 있어서, 상기 감지가능한 신호는 광이고, 상기 감지수단은 최소한 하나이상의 전자증배관으로 된 루미노미터로 된 것을 특징으로 하는 자동화 실험장치.11. An automated laboratory apparatus according to claim 10, wherein said detectable signal is light and said sensing means comprises a luminometer of at least one electron multiplier. 제12항에 있어서, 상기 혼합물은 다수개의 작은 웰내에 넣어지고, 상기 루미노미터는 상기 웰과 상기 전자증배관사이의 대응스캔을 형성하기 위한 스캔수단으로 됨으로써 전자증배관이 웰을 연속적으로 스캔하는 것을 특징으로 하는 자동화 실험장치.13. The method according to claim 12, wherein the mixture is placed in a plurality of small wells, and the luminometer serves as a scanning means for forming a corresponding scan between the well and the electron multiplier so that the electron multiplier continuously scans the well. Automated experimental apparatus, characterized in that. 최소한 하나이상의 미리 설정된 생체세포의 수집물을 생산 및 재생시키기에 적합한 규정된 조건을 인큐베이터내에 설정하는 스텝과, 상기 미리 설정된 수집물에서 인큐베이터의 세포를 인큐베이터내에 저장하는 스텝과, 규정된 조건하에 저장된 혼합물을 형성하도록 상기 세포와 혼합될 경우 상기 세포에서 감지가능한 변화를 발생하는 화학약품을 전달하며 인큐베이터에서 액체조작부로 화학약품을 로보트적으로 전달하는 스텝과, 혼합물을 형성하기 위하여 액체 조작부에서 화학약품을 세포와 혼합하는 스텝과, 인큐베이터내에서 배양을 위하여 혼합물을 인큐베이터로 로보트적으로 전달하는 스텝과, 상기 감지가능한 변화에 응답하여 감지기내에서 감지신호를 발생하는 스텝으로 이루어진 것을 특징으로 하는 실험방법.Establishing predetermined conditions in the incubator suitable for producing and reproducing at least one predetermined collection of biological cells, storing the cells of the incubator in the predetermined collection in the incubator, and stored under the prescribed conditions. Delivering a chemical that produces a detectable change in the cell when mixed with the cell to form a mixture, and robotically transferring the chemical from the incubator to the liquid control unit, and the chemical at the liquid control unit to form the mixture. And mixing the cells with the cells, robotically delivering the mixture to the incubator for culturing in the incubator, and generating a detection signal in the detector in response to the detectable change. . 최소한 하나 이상의 미리 설정된 생체세포의 수집물을 생산 및 재생시키기에 적합한 규정된 조건을 인큐베이터내에 설정하는 스텝과, 상기 미리 설정된 수집물에서 인큐베이터 셀내에 저장하는 스텝과, 규정된 조건하에 저장된 혼합물을 형성하도록 상기 세포와 혼합될 경우 상기 세포에서 감지가능한 변화를 발생하는 화학약품을 전달하며 인큐베이터에서 액체조작부로 세포를 전달하는 스텝과, 혼합물을 형성하기 위하여 액체조작부내에서 화학약품을 세포와 혼합하는 스텝과, 인큐베이터내에서 배양을 위하여 혼합물을 인큐베이터로 전달하는 스텝과, 인큐베이터내에서 배양후 혼합물을 감지기로 로보트적으로 전달하는 스텝과, 인큐베이터내에서 상기 감지가능한 변화에 응답하여 감지신호를 발생하는 스텝으로 이루어진 것을 특징으로 하는 자동화 실험장치.Establishing a predetermined condition in the incubator suitable for producing and regenerating a collection of at least one predetermined biological cell, storing in the incubator cell at the predetermined collection, and forming a mixture stored under the predetermined condition. Delivering a chemical that produces a detectable change in the cell when mixed with the cell and transferring the cell from the incubator to the liquid control unit, and mixing the chemical with the cell in the liquid control unit to form a mixture. And delivering the mixture to the incubator for incubation in the incubator, robotically delivering the mixture to the detector after incubation in the incubator, and generating a detection signal in response to the detectable change in the incubator. Characterized in that consists of Assimilation experiment apparatus. 최소한 하나이상의 미리 설정된 생체세포의 수집물을 생산 및 재생시키기에 적합한 규정된 조건을 인큐베이터내에 설정하는 스텝과, 상기 미리 설정된 수집물에서 인큐베이터 세포내에 저장하는 스텝과, 규정된 조건하에 저장된 혼합물을 형성하도록 상기 세포와 혼합될 경우 상기 세포에서 감지가능한 변화를 발생하는 화학약품을 전달하며 인큐베이터에서 액체조작부로 세포를 로보트적으로 전달하는 스텝과, 혼합물을 형성하기 위하여 액체 조작부에서 화학약품과 세포를 혼합하는 인큐베이터내에서 배양을 위하여 혼합물을 인큐베이터로 로보트적으로 전달하는 스텝으로 이루어진 것을 특징으로 하는 자동화 실험장치.Establishing predetermined conditions in the incubator suitable for producing and regenerating a collection of at least one predetermined biological cell, storing in the incubator cells in the predetermined collection, and forming a mixture stored under the prescribed conditions. Delivering a chemical that produces a detectable change in the cell when mixed with the cell, and robotically delivering the cell from the incubator to the liquid control unit, and mixing the chemical and the cell in the liquid control unit to form a mixture. Automated experimental device, characterized in that consisting of a step for robotically delivering the mixture to the incubator for incubation in the incubator. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019920700118A 1989-07-18 1990-07-18 Automated testing equipment KR920703785A (en)

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CA2063826A1 (en) 1991-01-19
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HUT61810A (en) 1993-03-01
EP0483226A4 (en) 1993-02-17

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