Temperature control apparatus and method

Abstract

An apparatus and method for performing automated sample preparation, DNA amplification and detection, which apparatus has heat-sinking, flat carriers for holding specimens and reagents, devices for heating and cooling and maintaining the specimen to or at any given temperature for any given time periods, and a computer to generate signals that control said temperatures and times.

What is claimed is: 
     
       1. An apparatus for providing temperature control to a specimen carrier, comprising: (a) one or more specimen carriers, each of said specimen carriers comprising a compartment for holding a specimen and reaction fluids;   (b) an enclosure having an interior space and having a site for positioning said one or more specimen carriers;   (c) a temperature-controlled plate located in said enclosure, said temperature-controlled plate having a first position in thermal contact with one side of said one or more specimen carriers placed at said site and a second position out of said thermal contact;   (d) means for moving said temperature-controlled plate between said first position and second position;   (e) means for providing laminar air flow between said temperature-controlled plate and said one or more specimen carriers when said temperature-controlled plate is in said second position;   (f) temperature control means capable of adjusting the temperature of said temperature-controlled plate in response to control signals; and   (g) computer means connected to said temperature control means and to said means for providing laminar air flow, wherein when said temperature-controlled plate is in the first position, the temperature of the one or more specimen carriers may be brought to a first temperature through thermal contact with the temperature-controlled plate and when the temperature-controlled plate is in the second position, laminar air flow provided between the temperature-controlled plate and the one or more specimen carriers may be used to bring the one or more specimen carriers to a second temperature.   
     
     
       2. An apparatus for providing temperature control to a specimen carrier according to claim 1, wherein the temperature-controlled plate is a heating plate. 
     
     
       3. An apparatus for providing temperature control to a specimen carrier according to claim 1, wherein the means for moving the temperature-controlled plate between the first and second positions comprises means for lifting the temperature-controlled plate into contact with the one or more specimen carriers and for lowering the temperature-controlled plate away from the one or more specimen carriers. 
     
     
       4. An apparatus for providing temperature control to a specimen carrier according to claim 1, wherein said compartment is formed by a carrier bottom and a cover, said cover movable between an open and closed position, and used for holding specimens and for treatment with reaction fluids. 
     
     
       5. An apparatus for providing temperature control to a specimen carrier according to claim 1, wherein the first temperature is a temperature which denatures nucleic acid complexes and the second temperature is chosen to be just below the melting temperature of primer oligonucleotides. 
     
     
       6. An apparatus for providing temperature control to a specimen carrier according to claim 1, further comprising sensing means in thermal contact with the temperature-controlled plate and connected to said computer means, wherein when a selected temperature is detected by said sensing means, said computer means causes the distance between the temperature-controlled plate and said one or more specimen carriers to change. 
     
     
       7. An apparatus for providing temperature control to a specimen carrier according to claim 6, wherein when said selected temperature is detected, laminar air flow between said temperature-controlled plate and said one or more specimen carriers is provided by said means for providing laminar air flow. 
     
     
       8. An apparatus for providing temperature control to a specimen carrier according to claim 1, wherein said means for providing said laminar air flow comprises a fan directing air through a plenum to a smaller space between said temperature-controlled plate and said one or more specimen carriers. 
     
     
       9. An apparatus for providing temperature control to a specimen carrier according to claim 1, wherein said temperature-control means is a part of said computer means. 
     
     
       10. A method for providing temperature control to one or more specimens in one or more specimen carriers in a specimen treatment process, comprising: (a) placing one or more specimen carriers in an enclosure having an interior space and having a site for positioning one or more specimen carriers, each of the said one or more specimen carriers comprising a compartment for holding a specimen;   (b) providing: (i) a temperature-controlled plate located in said enclosure, said temperature-controlled plate having a first position in thermal contact with one side of said one or more specimen carriers placed at said site and a second position out of said thermal contact;   (ii) means for moving said temperature-controlled plate between said first position and second position;   (iii) means for providing laminar air flow between said temperature-controlled plate and said one or more specimen carriers when said temperature-controlled plate is in said second position;   (iv) temperature control means capable of adjusting the temperature of said temperature-controlled plate in response to control signals; and   (v) computer means connected to said temperature control means and to said means for providing laminar air flow, wherein when said temperature-controlled plate is in the first position, the temperature of the one or more specimen carriers may be warmed through thermal contact the temperature-controlled plate and when the temperature-controlled plate is in the second position, laminar air flow provided between the one or more specimen carriers and the temperature-controlled plate may be used to cool the one or more specimen carriers;     (c) performing temperature-changing steps in a predetermined sequence along with one or more specimen treatment steps said temperature-changing steps including: (i) changing the temperature of said one or more specimen carriers to a warmer desired temperature by moving said temperature-controlled plate to said first position utilizing said means for moving said temperature-controlled plate between said first and second positions with respect to said site and providing control signals to adjust the temperature to said warmer desired temperature and to maintain the temperature of the temperature-controlled plate at said warmer desired temperature for a first predetermined time period; and   (ii) moving said temperature-controlled plate to said second position utilizing said means for moving the said temperature-controlled plate between said first and second positions and one ore more specimen carriers utilizing said means for providing laminar air flow to adjust the temperature of said temperature-controlled plate to a cooler desired temperature; and maintaining said cooler desired temperature for a second predetermined time period.     
     
     
       11. A method of providing temperature control to one or more specimens in one or more specimen carriers in a specimen treatment process according to claim 10, and wherein the sequence of the predetermined sequence of temperature-changing steps and the specimen treatment steps comprises: (a) warming the one or more specimen carriers to a temperature just above a gelling temperature of a selected matrix material before adding liquid matrix material to the one or more specimen carriers;   (b) after adding liquid matrix material containing a specimen having DNA to said one or more specimen carriers, cooling the one or more specimen carriers to lower than said gelling temperature;   (c) adding a treatment solution to the one or more specimen carriers and adjusting the temperature of the one or more specimen carriers to allow a desired treatment to occur;   (d) heating the one or more specimen carriers to 85° C. and maintaining g the temperature at about 85° C. until the gelled matrix material is dehydrated;   (e) denaturing the specimen DNA by saturating the dehydrated matrix material with distilled water and heating to about 95° C. for about 3-5 minutes;   (f) heating the one or more specimen carriers to about 72° C. and maintaining at about 72° C. while adding amplification reagents;   (g) adding amplification reagents to the one or more specimen carriers;   (h) initiating amplification thermal cycling after addition of amplification reagents, adjusting temperature to about 72° C. and maintaining at about 72° C. for about 2 minutes;   (i) heating the one or more specimen carriers to about 95° C. and maintaining at about 95° C. for about 20 seconds;   (j) repeating steps (h) and (i) about 24 times;   (k) cooling the one or more specimen carriers to about 72° C. and maintaining at about 72° C. for about 10 minutes; and   (l) washing fluids through each specimen earlier to remove unwanted materials and incubating at an incubation temperature. .Iadd.   
     
     
       12.  An apparatus for providing temperature control to one or more specimens, comprising: (a) one or more specimen carriers, each of said specimen carriers comprising a compartment for holding a specimen and reaction fluids not thicker than 0.5 millimeters;   (b) an enclosure having an interior space and having a site for positioning said one or more specimen carriers;   (c) a temperature-controlled plate located in said enclosure, said temperature-controlled plate capable of bringing said one or more specimen carriers to a first predetermined reaction temperature;   (d) means for bringing said one or more specimen carriers to a second predetermined reaction temperature;   (e) temperature control means capable of controlling the temperature of said temperature-controlled plate and adjusting the temperature of said one or more specimen carriers, in response to control signals; and   (f) computer means connected to said temperature control means and to said means for bringing said one or more specimen carriers to the second predetermined temperature wherein the amount of time said one or more specimen carriers is at either predetermined reaction temperature is repeatedly regulated, and wherein the temperature-controlled plate has a thermal mass enabling uniform heat transfer to and away from the specimens and the specimen carriers, wherein regulation of reaction temperature is sufficiently rapid to allow an amplification product to be obtained..Iaddend..Iadd.13. The apparatus for providing temperature control to one or more specimens according to claim 12, wherein the specimen carriers comprise microscope slides and a carrier assembly for receiving the slides as a carrier bottom and providing carrier edges and top pieces..Iaddend..Iadd.14. An apparatus for providing temperature control to one or more specimens, comprising:   (a) one or more specimen carriers, each of said specimen carriers comprising a compartment for holding a specimen and reaction fluids not thicker than 0.5 millimeters;   (b) an enclosure having an interior space and having a site for positioning said one or more specimen carriers;   (c) a temperature-controlled plate located in said enclosure, said temperature-controlled plate capable of bringing said one or more specimen carriers to a first predetermined reaction temperature;   .Iadd.(d) means for providing laminar air flow to bring said one or more specimen carriers to a second predetermined reaction temperature lower than the first predetermined reaction temperature;   .Iadd.(e) temperature control means capable of controlling the temperature of said temperature-controlled plate and adjusting the temperature of said one or more specimen carriers, in response to control signals; and   .Iadd.(f) computer means connected to said temperature control means and to said means for providing laminar air flow to bring said one or more specimen carriers to a second predetermined temperature lower than the first predetermined temperature, wherein the amount of time said one or more specimen carriers is at either predetermined reaction temperature is repeatedly regulated, and wherein the temperature-controlled plate has a thermal mass enabling uniform heat transfer to and away from the specimens and the specimen carriers, wherein regulation of reaction temperature is sufficiently rapid to allow an amplification product to be obtained..Iaddend..Iadd.15. The apparatus for providing temperature control to one or more specimens according to claim 13, wherein the specimen carriers comprise microscope slides and a carrier assembly for receiving the slides as a carrier bottom and providing carrier edges and top pieces..Iaddend.