EP0542422A1 - Plaque de microtitrage à puits multiples - Google Patents

Plaque de microtitrage à puits multiples Download PDF

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Publication number
EP0542422A1
EP0542422A1 EP92309172A EP92309172A EP0542422A1 EP 0542422 A1 EP0542422 A1 EP 0542422A1 EP 92309172 A EP92309172 A EP 92309172A EP 92309172 A EP92309172 A EP 92309172A EP 0542422 A1 EP0542422 A1 EP 0542422A1
Authority
EP
European Patent Office
Prior art keywords
well
microtiter tray
tray
liquid
wells
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.)
Withdrawn
Application number
EP92309172A
Other languages
German (de)
English (en)
Inventor
Harold R. Garner
Glen Shepherd
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.)
General Atomics Corp
Original Assignee
General Atomics Corp
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 General Atomics Corp filed Critical General Atomics Corp
Publication of EP0542422A1 publication Critical patent/EP0542422A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B01L3/50851—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates specially adapted for heating or cooling samples
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples

Definitions

  • the present invention generally pertains to multi-well microtiter trays which are useful for holding a plurality of minute liquid samples. More particularly, the present invention pertains to microtiter trays which can hold a plurality of liquid specimens while they are being cultivated by a temperature varying regimen and during periodic photometric or fluorometric measurements of the specimens. The present invention is particularly, but not exclusively, useful for DNA amplification and sequencing.
  • Microtiter trays with many wells for separately receiving liquid specimens have been used in the past to hold and store small specimens of sample material.
  • these earlier trays have been designed for use with robotics which are capable of engaging the microtiter tray to deposit the specimens in the wells of the tray.
  • the industry standard has been to provide microtiter trays which are separated by a center-to-center distance of nine millimeters.
  • the robotics which are used with currently available microtiter trays have been designed to comply with this standard. This restriction on spacing between the wells. of the tray can be perceived as a disadvantage if the desire is to process as many discrete samples as possible. There are, however, other potential disadvantages in presently available microtiter trays.
  • microtiter tray which can be used in processes which vary and maintain the temperature of the specimens according to predetermined temperature regimens, and which can be used during photometric or fluorometric analysis of the specimens.
  • an object of the present invention to provide a multi-well microtiter tray which is useful for simultaneously processing a high volume of minute liquid samples. Another object of the present invention is to provide a multi-well microtiter tray which is efficient in handling a very large number of individual minute liquid specimens. Yet another object of the present invention is to provide a multi-well microtiter tray which is useful with robotics which function in accordance with existing standards of the industry. Still another object of the present invention is to provide a multi-well microtiter tray which minimizes the storage space which is required for a very large number of minute liquid specimens. Another object of the present invention is to provide a multi-well microtiter tray which allows for substantially even heating of all liquid specimens being held in the tray.
  • a multi-well microtiter tray for use in a system, or in a method, for high volume processing of a plurality of minute liquid specimens includes a generally rectangular base having a substantially flat underside.
  • the top of the base is formed with a very large number of wells for holding a very large number of individual liquid specimens.
  • the top of the microtiter tray is formed with eight hundred and sixty four wells which are arranged in a rectangular array that is thirty-six by twenty-four.
  • Each well in the array has an opening and a bottom and is substantially cylindrical shaped to define a curved wall which tapers inwardly from the opening of the well to the bottom of the well.
  • the bottom of each well is distanced from the underside of the base and has an optical quality surface for transmitting light through the underside and through the liquid specimens held in the well.
  • the bottom of each well may be either flat, or rounded, or cone-shaped.
  • the system for using the multi-well microtiter tray includes a support structure which has a heat exchanger that is engageable with the microtiter tray.
  • the heat exchanger is a heat source which conductively transfers heat to the liquid specimens in the well through the base of the microtiter tray.
  • the heat exchanger acts as a heat sink and the support structure further includes a heating element.
  • this heating element may be a heat radiator such as would be the case for a microwave oven.
  • the heating element comprises a plurality of heating probes which can be inserted into selected wells of the microtiter tray to conduct heat through the tray to the other wells in the microtiter tray.
  • the system can also include a controller to coordinate the operation of the heating element with the operation of the heat exchanger to cycle the liquid specimens through predetermined temperature regimens.
  • microtiter tray Due to the construction of the base of the microtiter tray, light can be transmitted through the liquid specimens for photometrically or fluorometrically measuring the characteristics of the liquid specimens. This can be simultaneously done for all of liquid specimens held by the microtiter tray as they are being processed. Processes which can be accomplished using the system and methods for using a microtiter tray as intended for the present invention include DNA amplification sequencing and library construction.
  • the system for high volume processing a plurality of minute liquid specimens is shown and generally designated 10.
  • the system 10 includes a microtiter tray 12 which has a base 14. Formed into the top surface 16 of the base 14 are a plurality of wells 18.
  • base 14 of microtiter tray 12 is formed with eight hundred and sixty four (864) wells 18 which are arranged in a rectangular array that is thirty-six by twenty-four (36X24).
  • the center-to-center distances 20 and 22 between the rows and lines of the wells 18 are set at three millimeters (3mm).
  • base 14 is of a solid construction. Stated differently, there is no space between the wells 18. Instead, the material of base 14 fills what could otherwise be empty space between the wells 18. This construction helps provide temperature uniformity for all of the liquid specimens held in the wells 18 because base 14 establishes a shorter thermal path from a well 18 to another well 18 to facilitate heat transfer therebetween.
  • microtiter tray 12 is made of an optically clear rigid plastic such as polystyrene, polypropelene, or polycarbonate.
  • the plastic of tray 12 can include carbon fibers, alumina, aluminum oxide, or other metals which will enhance the thermal conductivity of the tray 12 but which will not interfere with its biological performance.
  • the tray 12 can be manufactured by any process well known in the pertinent art, such as by injection molding.
  • FIG. 1 also shows that microtiter tray 12 has an overhang 24 which forms a detent 26 between the overhang 24 and the base 14 of tray 12. Additionally, a pair of registration points 28a and 28b can be formed into top surface 16 of tray 12 in order to align the tray 12 with a robot (not shown in Figure 1) for purposes to be discussed below. Also, the top surface 16 of microtiter tray 12 has a flat area 30 and a flat area 32 on which an identification strip, such as bar code 33, can be placed. As will be appreciated by the skilled artisan, some ability to identify the microtiter tray 12 and the liquid specimens held in the wells 18 of the tray 12 can be essential.
  • the system 10 of the present invention includes a support structure on which microtiter tray 12 can be placed.
  • the support structure may be merely a storage shelf.
  • the support structure may be part of a library for biological specimens", such as a shelf in a standard refrigerator (not shown) or an incubator (not shown).
  • the support structure may encompass any platform where the microtiter tray 12, and the liquid specimens in the wells 18 of tray 12, can be stored for prolonged periods of time.
  • a more operational function is for heat transfer with the liquid specimens.
  • the support structure is as a heat exchanger 34.
  • heat exchanger 34 can be either a heat source or a heat sink. In either case, it must be operatively engageable with the microtiter tray 12.
  • heat exchanger 34 can have a surface 36 which will directly abut against the flat underside 38 of base 14 of the microtiter tray 12.
  • a thermal grease, or some other such substance well known in the pertinent art can be placed on the surface 36 to facilitate heat transfer between the heat exchanger 34 and the microtiter tray 12.
  • thermoelectric tray 12 An additional feature of the system 10 of the present invention which promotes good heat transfer characteristics for the microtiter tray 12 is that both the top surface 16 and underside 38 of the tray 12 are flat and each have a relatively large area.
  • heat exchanger 34 can be formed with a ridge 40 which is matingly received into the detent 26 of microtiter tray 12 to help hold the tray 12 in position on heat exchanger 34.
  • the exchanger 34 can include controls 42a,b for the purpose of precisely controlling the amount of heat which is to be applied from the heating coils 44 of heat exchanger 34 to the tray 12 during the processing of the liquid specimens in the wells 18.
  • each of the wells 18 in microtiter tray 12 is essentially a hollow cylindrical recess which is formed into the top surface 16 of the tray 12.
  • each well 18 has an opening 46 and is defined by a cylindrical sidewall 48 which extends from the opening 46 to the bottom of the well.
  • a well 18 is shown to have a substantially flat bottom 50, whereas in Figure 2B the well 18 has a rounded bottom 52, and in Figure 2C the well 18 is shown with a cone shaped bottom 54.
  • a particular shape for the bottom of well 18 may be preferable.
  • the rounded bottom 52 of well 18 is better suited for use with liquid specimens which include cells which might tend to cling to the angled surfaces presented by a flat surface 50.
  • the cone-shaped surface 54 might be preferable for use with automatic pipetting.
  • the bottoms 50, 52, and 54 of wells 18, as well as underside 38 of base 14, must be optically clear. Stated differently, sufficient light with which to make photometric or fluorometric measurements of the liquid specimens 56 being held in the well 18 must be able to pass through underside 38, through the bottom 50, 52, 54 of well 18 and through the liquid specimen held in well 18.
  • optically clear means that the bottom 50, 52, or 54 has a smoothness such that deviations in their surface do not exceed more than one quarter wavelength of the light which is passed through tray 12 and the liquid specimen 56 to make photometric or fluorometric measurements of the liquid specimen 56.
  • the light used for photometric or fluorometric measurements of the liquid specimen 56 will be primarily in the visual range.
  • the preferred dimensions for a typical well 18 will be best appreciated with specific reference to Figure 2D.
  • center-to-center distance 20 between adjacent wells 18 along either a line or a row of wells 18 is approximately three millimeters (3mm).
  • a standard tool having a nine millimeter (9mm) spacing between probes can be used with the 3mm spacing of tray 12 if it is repeatedly used three times. This can be carried further.
  • the center-to-center distance can be four and a half millimeters (4.5mm).
  • the standard robot would then need to be used twice. It happens that several configurations of the tray 12 can be used with standard robotics just so long as the center-to-center spacing is equal to nine divided by n, where n is an integer. Obviously, the volume capacity of the wells 18 will be sacrificed as more wells 18 and incorporated into the tray 12.
  • the diameter 58 of the opening 46 is approximately two millimeters (2mm).
  • Figure 2D shows that the sidewalls 48 of well 18 are tapered inwardly from the opening 46 of well 18 to its bottom 50.
  • the angle of taper 60 is approximately one degree (1°).
  • the depth 62 of well 18 is approximately seven millimeters (7mm) with the result that well 18 is dimensioned to hold a liquid specimen 56 having a volume of approximately twenty microliters (20 ul).
  • the actual volume of the liquid specimen 56 which is to be deposited into a well 18 may have a liquid volume that is more on the order of one or two microliters (1-2 ul).
  • liquid specimens which have total volumes on the order of one or two microliters can be hard to handle.
  • a specific problem confronted by the present invention involves the transfer of such a small liquid specimen 56 into a well 18 of the microtiter tray 12. This problem stems from the fact that surface tension of the liquid specimen 56 will, at this level, be sufficient to prevent the specimen from forming into a drop which will fall into the well 18. Consequently, to assist the transfer of the liquid specimen 56 into the well 18, the sidewalls 46 of the well 18 are tapered.
  • a liquid specimen 56 will be transferred from a source of the liquid (not shown) to the well 18' of microtiter tray 12 by a device such as the probe 66.
  • the size of the liquid specimen 56 may be so small that it cannot form into a drop which will fall into the well 18'.
  • probe 66 can be made to contact the sidewall 48' when it is inserted into the well 18'. This contact, a mere kiss if you will, is sufficient to wet the sidewall 48' with the liquid specimen 56 and cause the liquid specimen 56 to flow into the bottom 50' of the well 18'.
  • a robot 68 may be used to transfer liquid specimens 56 form a source of the specimens (not shown) to the microtiter tray 12.
  • the robot 68 is outfitted with an end effector 70 which has a plurality of probes 66 (e.g eight hundred and sixty four) for this purpose.
  • Each of the probes 66 is designed, in a manner well known in the pertinent art, to carry a minute quantity of a liquid specimen 56 for deposit on the tray 12.
  • the robot 68 can include well known means (not shown) on end effector 70 which will register with the registration points 28a,b on microtiter tray to properly align the probes 66 on end effector 70 with the wells 18 on microtiter tray 12.
  • robot 68 is operable to engage end effector 70 with the microtiter tray 12 to deposit liquid specimens 18 in the wells 18 of tray 12.
  • heat exchanger 34 can be operated to heat the specimens 56 as desired by the user. This, of course, is for the embodiment of the system 10 wherein the heat exchanger 34 acts as a heat source. According to the present invention, various temperature regimens may be followed by the heat exchanger 34 when heating the specimens 56. DNA amplification is but one example of such a regimen.
  • microtiter tray 12 is intended to be used during photometric or fluorometric measurements of the liquid specimens 56 which are deposited into the wells 18 of the tray 12.
  • a light source 72 is schematically shown in a relationship with the tray 12 for making such photometric or fluorometric measurements.
  • the light source 72 may be of any type well known in the pertinent art, but it is preferably of a type which will emit a light 74 in the visual range having identifiable wavelengths, for example ⁇ 1. According to standard photometric or fluorometric analysis a resultant light 76 having a different wavelength, for example ⁇ 2, will result due to the particular characteristics of the liquid specimen 56.
  • This resultant light 76 will then be received at a light detector 78 and the wavelength difference between light 74 and resultant light 76 can be used to measure the characteristics of the liquid specimen 56.
  • a photometric or fluorometric analysis of the liquid specimens 56 in microtiter tray 12 can be made at any time.
  • the heat exchanger 34 can be used as a heat sink as well as a heat source. Accordingly, when heat exchanger 34 is to a heat sink, another source of heat is needed for procedures wherein the liquid specimens 56 are to be cycled through a temperature regimen.
  • Figure 4 indicates that heat exchanger 34 can be incorporated as part of an oven 80 and that the microtiter tray 12 can be positioned on the exchanger 34 in oven 80. The door 82 of oven 80 can then be shut (not shown) to enclose microtiter tray 12 with its liquid specimens 56 inside the oven 80. Both the heating function of the oven 80, and the cooling function of the heat sink exchanger 34, can then be controlled from the control panel 84 to cycle the liquid specimens 56 in tray 12 through any temperature regimen desired by the user.
  • oven 80 may be of any type well known in the pertinent art.
  • oven 80 may be a microwave oven or it may be an air thermal cycler.
  • the system 10 of the present invention may also include a heat transfer tool 86.
  • the heat transfer tool 86 comprises a block member 88 on which are mounted a plurality of heat probes 90 that extend through the block member 88.
  • the heat probes 90 each have and extension 92 which extends upwardly from the block member 88 and an engager 94 which extends downwardly from the block member 88.
  • the heat probes 90 are mounted on the block member 88 of heat transfer tool 86 so that when the tool 86 is engaged with a microtiter tray 12 an engager 94 is inserted into only every other one of the wells 18.
  • the probes 90 may be spaced more than one well 18 apart from each other. This alternating engagement between the heat probes 90 of heat transfer tool 86 and the wells 80 of tray 12 is followed along both the lines and rows of the array of wells 18 in microtiter tray 12.
  • Figure 6 also indicates that a thermal grease 96 can be placed on the engagers 94 to assist in the transfer of heat from each of the heat probes 90 to the liquid specimens 56 in adjacent wells 18.
  • Figures 7 and 8 show yet another embodiment for the system 10 of the present invention wherein heat exchanger 34 is used as a heat sink.
  • the heating element for transferring heat to the liquid specimens 56 in wells 18 of the microtiter tray 12 incorporates a robot 98 which operatively maneuvers an end effector 100.
  • the end effector 100 includes a plurality of probes 102 which are each engageable with one of the wells 18 of microtiter tray 12 substantially as shown in Figure 8. Similar to the arrangement disclosed above for heat transfer tool 86, the probes 102 of end effector 100 are spaced to be engageable with alternate wells 18 of the tray 18.
  • this alternating arrangement extends along both the lines and the rows of wells 18 in tray 12.
  • the ability of the probes 102 to transfer heat from the probes 102 to the tray 12, and eventually to the liquid specimens 56 in the adjacent wells 18, will be enhanced by the use of a thermal grease 104 which can be placed on each of the probes 102.
  • Figure 7 shows that a control box 106 is connected with heat exchanger 34 through a line 108, and that the control box is connected with robot 96 through a line 110.
  • the control box 106 can be used to establish the temperature of the heating probes 102.
  • the control box 106 can be used to control heat sink 34 for removing heat from the specimens 56 in tray 12. Consequently, as with the other embodiments of the system 10 of the present invention, the embodiment shown in Figures 7 and 8 can be used to cycle the temperature of the liquid specimens 56 in the wells 18 of microtiter tray 12 according to any predetermined temperature regimen desired by the operator.
  • photometric or fluorometric measurements can be made as desired. Specifically, the general procedure discussed above with reference to Figure 2A can be accomplished whenever required by the particular process being followed.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Hematology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
EP92309172A 1991-11-12 1992-10-08 Plaque de microtitrage à puits multiples Withdrawn EP0542422A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US79036591A 1991-11-12 1991-11-12
US790365 1991-11-12

Publications (1)

Publication Number Publication Date
EP0542422A1 true EP0542422A1 (fr) 1993-05-19

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