(12) United States Patent
`Brown et al.
`
`(S4) FLEXIBLE HEATING COVER ASSEMBLY
`FOR THERMAL CYCLING OF SAMPLES OF
`BIOLOGICAL MATERIAL
`
`(7S)
`
`Inventors: Larry R. Brown, Carlsbad, CA (US);
`William D. Brumley, Vista, CA (US);
`Kenneth J. Zajac, San Diego, CA (US)
`(73) Assignee: Stratagene, La Jolla, CA (US)
`( * ) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 3S
`U.S.C. 1S4(b) by 0 days.
`
`(21) Appl. No. : 10/262,994
`(22) Filed:
`(6S)
`
`Oct. 2, 2002
`
`Prior Publication Data
`
`(Sl)
`
`(S2)
`
`(S8)
`
`(S6)
`
`US 2004/0065655 Al Apr. 8, 2004
`
`Int. Cl.7 ........................... C12M 1/00; C12M 1/38;
`HOSE 3/20; HOSE 3/34; GOSD 23/20
`U.S. Cl. ....................... 219/428; 219/38S; 219/S21;
`219/S30; 219/S3S; 219/S49; 43S/286.1;
`43S/288.4; 43S/304.4
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219/38S, 428,
`Field of Search
`219/429, 430, 436, S21, S28, S30, S3S,
`S49; 43S/289.1, 288.4, 286.1, 30S.3, 30S.4
`
`References Cited
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`. . . . . . . . . . . . . . . . 165/80 B
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`. . . . . . . . . . . . . . . 435/290
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`. . . . . . . . . . . . . 364/500
`Moser et al.
`. . . . . . . . . . . . . . . 422/104
`
`I lllll llllllll Ill lllll lllll lllll lllll lllll 111111111111111111111111111111111
`US006730883B2
`US 6, 730,883 B2
`
`(10) Patent No.:
`(4S) Date of Patent:
`
`May 4, 2004
`
`5,656,493 A
`5,710,381 A
`5,779,981 A
`5,785,926 A
`
`. . . . . . . . . . . . 435/286.l
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`1/1998 Atwood et al.
`. . . . . . . . . . 73/864.91
`7/1998 Danssaert et al.
`. . . . . . . . . . . . 422/99
`7/1998 Seubert et al.
`. . . . . . . . . . . . . . 422/100
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`(List continued on next page.)
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`EP
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`438883
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`07-308183
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`WO 98/43740
`WO 00/32312
`
`7 /1991
`6/1992
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`7/1993
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`12/1997
`12/1989
`10/1998
`6/2000
`
`BOlD/1/30
`Cl 2Q/l/68
`
`. . . . . . . . . . . . Cl2M/l/00
`. . . . . . . . . . . . Cl2M/l/00
`. . . . . . . . . . . . Cl2M/l/00
`. . . . . . . . . . . . . BOlL/7/00
`. . . . . . . . . . . . . BOlL/7/00
`. . . . . . . . . . . . . BOlL/7/00
`
`OTHER PUBLICATIONS
`circulator," KR
`"Multi-functional
`temperature
`2001038297 A, abstract and single figure, May lS, 2001. *
`Primary Examiner-Joseph Pelham
`(74) Attorney, Agent, or Firm-Palmer & Dodge, LLP;
`Kathleen M. Williams; David J. Dykeman
`(S7)
`
`ABSTRACT
`
`A flexible heating cover assembly for an apparatus for
`heating samples of biological material with substantial tem­
`perature uniformity includes a housing having a plurality of
`engageable enclosure components; a resistive heater having
`a plurality of heater element areas; a heater backing plate
`providing stability to the resistive heater; a force distribution
`system that distributes a force over the heater backing plate;
`and a support plate providing stiffness for the force distri­
`bution system, wherein the arrangement of the resistive
`heater, the heater backing plate, the force distribution system
`and the support plate provide substantial temperature uni­
`formity among a plurality of sample tubes for receiving
`samples of biological material. The flexible heating cover
`assembly improves the uniformity, efficiency, quality, reli­
`ability and controllability of the thermal response during
`thermal cycling of the biological material.
`
`82 Claims, 28 Drawing Sheets
`
`308
`
`THERMO FISHER EX. 1022
`
`

`
`US 6, 730,883 B2
`Page 2
`
`U.S. PATENT DOCUMENTS
`
`5,813,233 A
`5,819,842 A
`5,849,208 A
`5,851,492 A
`5,928,907 A
`6,004,512 A
`6,054,263 A
`
`................. 62/3.7
`9/1998 Okuda et al.
`................ 165/206
`10/1998 Potter et al.
`12/1998 Hayes et al. .................. 216/94
`* 12/1998 Blattner ................... 435/288.4
`..... 435/91. 2
`7/1999 Woudenberg et al.
`12/1999 Titcomb et al. ............... 422/63
`. ... ... ... ... 435/4
`4/2000 Danssaert et al.
`
`6,093,370 A
`6,106,784 A
`6,153,426 A
`6,337,435 Bl
`6,638,761 B2 *
`6,657,169 B2 *
`
`7/2000
`8/2000
`* 11/2000
`1/2002
`10/2003
`12/2003
`
`............. 422/68.1
`Yasuda et al.
`................. 422/104
`Lund et al.
`................ 435/305.4
`Heimberg
`Chu et al. .. ... ... ... ... .. ... 136/242
`............... 435/288.4
`Shin et al.
`....................... 219/385
`Brown
`
`* cited by examiner
`
`THERMO FISHER EX. 1022
`
`

`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 1 of 28
`
`US 6, 730,883 B2
`
`200
`\
`
`FIG. 1
`
`THERMO FISHER EX. 1022
`
`

`
`u .s. Patent
`
`May 4, 20M
`
`Sheet 2 of 28
`
`US 6,730,883 B2
`
`200
`""'"
`
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`
`THERMO FISHER EX. 1022
`
`

`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 3 of 28
`
`US 6, 730,883 B2
`
`/'200
`
`128
`
`128
`
`FIG. 3
`
`THERMO FISHER EX. 1022
`
`

`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 4 of 28
`
`US 6, 730,883 B2
`
`-LL
`
`THERMO FISHER EX. 1022
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 5 of 28
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`US 6, 730,883 B2
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`U.S. Patent
`
`May 4, 2004
`
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`May 4, 2004
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`May 4, 2004
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`May 4, 2004
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`May 4, 2004
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`May 4, 2004
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`May 4, 2004
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`US 6, 730,883 B2
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`U.S. Patent
`
`May 4, 2004
`
`Sheet 15 of 28
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`US 6,730,883 B2
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`May 4, 2004
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 17 of 28
`
`US 6, 730,883 B2
`
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`308
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 18 of 28
`
`US 6, 730,883 B2
`
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 19 of 28
`
`US 6, 730,883 B2
`
`308
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`310
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 20 of 28
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`US 6,730,883 B2
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`U.S. Patent
`
`May 4, 2004
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`Sheet 21 of 28
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`US 6, 730,883 B2
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`U.S. Patent
`
`May 4, 2004
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`Sheet 22 of 28
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`US 6,730,883 B2
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`U.S. Patent
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`May 4, 2004
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`Sheet 23 of 28
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`US 6, 730,883 B2
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 24 of 28
`
`US 6, 730,883 B2
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 25 of 28
`
`US 6, 730,883 B2
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`U.S. Patent
`
`May 4, 2004
`
`Sheet 26 of 28
`
`US 6, 730,883 B2
`
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 27 of 28
`
`US 6, 730,883 B2
`
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`
`U.S. Patent
`
`May 4, 2004
`
`Sheet 28 of 28
`
`US 6, 730,883 B2
`
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`
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`
`

`
`US 6, 730,883 B2
`
`1
`FLEXIBLE HEATING COVER ASSEMBLY
`FOR THERMAL CYCLING OF SAMPLES OF
`BIOLOGICAL MATERIAL
`
`RELATED APPLICATIONS
`
`None.
`
`FIELD OF THE INVENTION
`
`The present invention relates to a heating cover assembly
`for an apparatus for heating samples of biological material,
`and more particularly to a flexible heating cover assembly
`that improves the uniformity, efficiency, quality, reliability
`and controllability of the thermal response during thermal 15
`cycling of DNA samples to accomplish a polymerase chain
`reaction, a quantitative polymerase chain reaction, a reverse
`transcription-polymerase chain reaction, or other nucleic
`acid amplification types of experiments.
`
`BACKGROUND OF THE INVENTION
`
`Techniques for thermal cycling of DNA samples are
`known in the art. By performing a polymerase chain reaction
`(PCR), DNA can be amplified. It is desirable to cycle a
`specially constituted liquid biological reaction mixture
`through a specific duration and range of temperatures in
`order to successfully amplify the DNA in the liquid reaction
`mixture. Thermal cycling is the process of melting DNA,
`annealing short primers to the resulting single strands, and
`extending those primers to make new copies of double
`stranded DNA The liquid reaction mixture is repeatedly put
`through this process of melting at high temperatures and
`annealing and extending at lower temperatures.
`In a typical thermal cycling apparatus, a biological reac­
`tion mixture including DNA will be provided in a large
`number of sample wells on a thermal block assembly. It is
`desirable that the samples of DNA have temperatures
`throughout the thermal cycling process that are as uniform
`as reasonably possible. Even small variations in the tern- 40
`perature between one sample well and another sample well
`can cause a failure or undesirable outcome of the experi­
`ment. For instance, in quantitative PCR, one objective is to
`perform PCR amplification as precisely as possible by
`increasing the amount of DNA that generally doubles on 45
`every cycle; otherwise there can be an undesirable degree of
`disparity between the amount of resultant mixtures in the
`sample wells. If sufficiently uniform temperatures are not
`obtained by the sample wells, the desired doubling at each
`cycle may not occur. Although the theoretical doubling of 50
`DNA rarely occurs in practice, it is desired that the ampli­
`fication occurs as efficiently as possible.
`In addition, temperature errors can cause the reactions to
`improperly occur. For example, if the samples are not
`controlled to have the proper annealing temperatures, certain
`forms of DNA may not extend properly. This can result in
`the primers in the mixture annealing to the wrong DNA or
`not annealing at all. Moreover, by ensuring that all samples
`are uniformly heated, the dwell times at any temperature can
`be shortened, thereby speeding up the total PCR cycle time.
`By shortening this dwell time at certain temperatures, the
`lifetime and amplification efficiency of the enzyme are
`increased. Therefore, undesirable temperature errors and
`variations between the sample well temperatures should be
`decreased.
`Prior art heating covers used in PCR heating equipment
`are simple, stiff, and relatively inexpensive. The prior art
`
`2
`designs have mainly involved a stiff metal plate, a simple
`resistive heater, and an insulating cover. Because quantita­
`tive data was not generated, the heating covers did not have
`to control condensation in the biological samples as pre-
`s cisely as the heating covers used in QPCR equipment. Also,
`because optical data was not collected, the prior art heating
`cover designs were not complicated with the need to provide
`a means to excite and collect the optical data through the
`heating cover. Prior art heating covers used in QPCR heating
`10 equipment are mainly derived from their earlier PCR coun­
`terparts that provide a means for optical signal transmission,
`but, prior art heating covers are still mainly stiff designs
`which do not provide a uniform force distribution about the
`sample containers.
`Prior art heating covers are difficult to use, expensive,
`complicated and do not provide uniform thermal contact or
`uniform force distribution about the sample wells. U.S. Pat.
`No. 5,475,610 discloses an instrument for performing PCR
`employing a cover which can be raised or lowered over a
`20 sample block. U.S. Pat. No. 5,475,610 does not disclose a
`cover assembly that is flexible to provide a more uniform
`thermal contact and force distribution on the sample tube
`caps. U.S. Pat. No. 5,928,907 discloses a system for carrying
`out real time fluorescence-based measurements of nucleic
`25 acid amplification products. U.S. Pat. No. 5,928,907 does
`not disclose a cover assembly that is flexible to provide a
`more uniform thermal contact and force distribution on the
`sample tube caps. The prior art does not disclose a cover
`assembly that is flexible to provide a more uniform thermal
`contact and force distribution on the sample tube caps.
`In light of the foregoing, there is a need in the art for a
`flexible heating cover assembly that enhances the thermal
`response uniformity, efficiency, quality, reliability and con­
`trollability of the DNA sample wells in the thermal cycling
`apparatus.
`
`30
`
`35
`
`SUMMARY OF THE INVENTION
`
`The present invention is a flexible heating cover assembly
`that improves the uniformity, efficiency, quality, reliability
`and controllability of the thermal response during thermal
`cycling of DNA samples to accomplish a polymerase chain
`reaction, a quantitative polymerase chain reaction, a reverse
`transcription-polymerase chain reaction, or other nucleic
`acid amplification types of experiments.
`The present invention is a flexible heating cover assembly
`for an apparatus for heating samples of biological material
`with substantial temperature uniformity including a housing
`having a plurality of engageable enclosure components; a
`resistive heater located within the housing, the resistive
`heater including a plurality of heater element areas; a heater
`backing plate engaging the resistive heater and providing
`protection and stability to the resistive heater; a force
`distribution system that engages the heater backing plate and
`ss distributes a force over the heater backing plate; and a
`support plate providing stiffness for the force distribution
`system, wherein the arrangement of the resistive heater, the
`heater backing plate, the force distribution system and the
`support plate provide substantial temperature uniformity
`60 among a plurality of sample tubes for receiving samples of
`biological material. The flexible heating cover assembly
`improves the uniformity, efficiency, quality, reliability and
`controllability of the thermal response during thermal
`cycling of DNA samples.
`In another aspect of the present invention, the resistive
`heater produces a non-uniform heat distribution along a
`surface exposed to the plurality of sample tubes. The resis-
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`3
`tive heater further comprises a plurality of heater element
`areas including at least one outer heater element area and at
`least one central heater element area.
`In another aspect of the present invention, the heater
`backing plate is thin to promote flexibility when the heater 5
`backing plate is connected to the resistive heater. The heater
`backing plate is composed of a thermally conductive mate­
`rial.
`In another aspect of the present invention, the force
`distribution system further comprises at least one spring 10
`strip and a spring retainer plate. The at least one spring strip
`has an elongated body and a plurality of spring extensions to
`distribute the force uniformly on the heater backing plate.
`In another aspect of the present invention, the support
`plate has sufficient stiffness to provide a reaction force for
`the force distribution system with minimal deflection of the
`support plate.
`In another aspect of the present invention, the resistive
`heater, the heater backing plate, and the support plate each
`comprise a plurality of aligned sample well openings, each
`sample well opening corresponding to a respective sample
`tube of the plurality of sample tubes.
`The present invention is a flexible heating cover assembly
`with enhanced functions including the flexibility of the 25
`cover assembly and the force distribution. In addition, the
`flexible heating cover assembly of the present invention
`enables the resistive heater to float in a vertical direction, so
`that the resistive heater has some freedom of movement
`vertically which leads to a more uniform thermal contact and 30
`force distribution and more accurate and consistent results.
`The flexible heating cover assembly of the present invention
`provides thermal insulation for the upper portion of the
`sample tubes and the sample caps.
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`4
`FIG. 10 is a perspective view of a heat sink of a thermal
`system base.
`FIG. 11 is a bottom view of a heat sink of a thermal
`system base.
`FIG. 12 is a top view of a solid state heater a heat sink of
`a thermal system base.
`FIG. 13 is a side view of a solid state heater a heat sink
`of a thermal system base.
`FIG. 14 is a perspective view of a solid state heater of a
`thermal system base.
`FIG. 15 is a top view of a spacer bracket with a solid state
`heater of a thermal system base.
`FIG. 16 is a top perspective view of a spacer bracket of
`15 a thermal system base.
`FIG. 17 is a bottom perspective view of a spacer bracket
`of a thermal system base.
`FIG. 18 is a top view of a heat sink, a bottom resistive
`20 heater, and a plurality of solid state heaters of a thermal
`system base.
`FIG. 19 is a bottom view of a thermal block plate and a
`plurality of solid state heaters of a thermal system base.
`FIG. 20 is a top exploded assembly view of a flexible
`heating cover assembly of the present invention showing
`how a stiff support plate, a spring strip, a spring retainer
`plate, a heater backing plate, a plurality of heater slides, a
`resistive heater, a cover assembly skirt interact with a
`plurality of biological sample tubes having sample caps.
`FIG. 21 is a bottom exploded assembly view of a flexible
`heating cover assembly of the present invention showing
`how a stiff support plate, a spring strip, a spring retainer
`plate, a heater backing plate, a plurality of heater slides, a
`resistive heater, a cover assembly skirt interact with a
`35 plurality of biological sample tubes having sample caps.
`FIG. 22 is a perspective view of a resistive heater of a
`flexible heating cover assembly of the present invention
`showing a layout of a plurality of heater element areas.
`FIG. 23 is a top perspective view of a resistive heater of
`a flexible heating cover assembly of the present invention
`showing a thermistor.
`FIG. 24 is a bottom perspective view of a resistive heater
`of a flexible heating cover assembly of the present invention
`showing a plurality of insulating pads.
`FIG. 25 is a top view of a resistive heater of a flexible
`heating cover assembly of the present invention showing a
`thermistor.
`FIG. 26 is a side view of a resistive heater of a flexible
`50 heating cover assembly of the present invention.
`FIG. 27 is a perspective view of a heater backing plate of
`a flexible heating cover assembly of the present invention.
`FIG. 28 is a top view of a heater backing plate of a flexible
`heating cover assembly of the present invention.
`FIG. 29 is a top perspective view of a resistive heater
`engaging a heater backing plate of a flexible heating cover
`assembly of the present invention.
`FIG. 30 is a bottom perspective view of a resistive heater
`engaging a heater backing plate of a flexible heating cover
`assembly of the present invention.
`FIG. 31 is a bottom view of a resistive heater engaging a
`heater backing plate of a flexible heating cover assembly of
`the present invention.
`FIG. 32 is a side view of a resistive heater engaging a
`heater backing plate of a flexible heating cover assembly of
`the present invention.
`
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`55
`
`The accompanying drawings, which are incorporated in
`and constitute a part of this specification, illustrate several
`embodiments of the invention and together with the
`description, serve to explain the principles of the invention. 40
`The present invention will be further explained with refer­
`ence to the attached drawings, wherein like structures are
`referred to by like numerals throughout the several views.
`The drawings shown are not necessarily to scale, with
`emphasis instead generally being placed upon illustrating 45
`the principles of the present invention.
`FIG. 1 is a top perspective view of a flexible heating cover
`assembly of the present invention.
`FIG. 2 is a bottom perspective view of a flexible heating
`cover assembly of the present invention.
`FIG. 3 is a perspective view of a flexible heating cover
`assembly of the present invention attached to an apparatus
`for thermally cycling samples of a biological material.
`FIG. 4 is a front sectional view of a flexible heating cover
`assembly of the present invention attached to an apparatus
`for thermally cycling samples of a biological material.
`FIG. 5 is a partial enlarged front sectional view of a
`flexible heating cover assembly of the present invention.
`FIG. 6 is a top view of a thermal block assembly of a 60
`thermal system base.
`FIG. 7 is a perspective view of a thermal block assembly
`of a thermal system base.
`FIG. 8 is a perspective sectional view of a sample well of
`a thermal system base.
`FIG. 9 is a perspective view of a sensor cup of a thermal
`system base.
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`5
`FIG. 33 is a perspective view of a spring strip of a flexible
`heating cover assembly of the present invention.
`FIG. 34 is a top view of a spring strip of a flexible heating
`cover assembly of the present invention.
`FIG. 35 is a side view of a spring strip of a flexible heating
`cover assembly of the present invention.
`FIG. 36 is a perspective view of a spring retainer plate of
`a flexible heating cover assembly of the present invention.
`FIG. 37 is a top view of a spring retainer plate of a flexible
`heating cover assembly of the present invention.
`FIG. 38 is a top perspective view of a stiff support plate
`of a flexible heating cover assembly of the present invention.
`FIG. 39 is a bottom perspective view of a stiff support
`plate of a flexible heating cover assembly of the present 15 requirements. For example, the sample wells could be
`
`6
`plate 22 is substantially rectangular and is of sufficient size
`to accommodate a plurality of sample wells 24 on the top
`surface, but could be of other shapes (i.e., circular, oval,
`square). In the embodiment shown in the drawings, the plate
`5 22 accommodates 96 sample wells 24 in a grid having eight
`columns and twelve rows. The sample wells 24 are in an 8
`by 12 grid with center-to-center spacing between adjacent
`sample wells 24 of about nine millimeters. In other embodi­
`ments of the present invention, there may be more or less
`10 than 96 sample wells, the sample well arrangement may
`vary, and the center-to-center measurement between adja­
`cent sample wells 24 may be more or less than nine
`millimeters. It is to be understood that the number of sample
`wells can be varied depending on the specific application
`
`invention.
`FIG. 40 is a perspective view of a heater slide of a flexible
`heating cover assembly of the present invention.
`FIG. 41 is a front view of a heater slide of a flexible
`heating cover assembly of the present invention showing the
`U-shape of the preferred heater slide.
`While the above-identified drawings set forth preferred
`embodiments of the present invention, other embodiments
`of the present invention are also contemplated, as noted in
`the discussion. This disclosure presents illustrative embodi­
`ments of the present invention by way of representation and
`not limitation. Numerous other modifications and embodi­
`ments can be devised by those skilled in the art which fall
`within the scope and spirit of the principles of the present
`invention.
`
`DETAILED DESCRIPTION
`
`A flexible heating cover assembly of the present invention
`is illustrated generally at 200 in FIGS. 1 and 2. As best
`shown in FIGS. 20 and 21, the flexible heating cover
`assembly 200 includes a cover assembly skirt 250, a resis­
`tive heater 300, a heater backing plate 350, a spring strip
`400, a spring retainer plate 450, a stiff support plate 500, and
`a plurality of heater slides 550. The flexible heating cover
`assembly 200 engages a plurality of biological sample tubes
`140 having sample caps 146.
`As shown in FIG. 3, the flexible heating cover assembly
`200 can be attached to an apparatus for thermally cycling
`samples of a biological material. The flexible heating cover
`assembly 200 can be attached to any apparatus for thermal
`cycling of DNA samples to accomplish a polymerase chain
`reaction, a quantitative polymerase chain reaction, a reverse
`transcription-polymerase chain reaction, or other nucleic
`acid amplification types of experiments. For example, the
`flexible heating cover assembly 200 can be attached to the
`apparatus for thermally cycling samples of a biological
`material disclosed in assignee's co-pending U.S. patent
`application Ser. No. 09/364,051, the entirety of which is
`hereby incorporated by reference. When combined with a
`thermal system base 15 (which contains a thermal block
`assembly 20 for accepting samples and means to heat and
`cool the thermal block assembly 20), the flexible heating
`cover assembly 200 improves the quality of the thermal
`response of the system for quantitative PCR.
`The thermal system base 15 includes a plurality of sample
`wells for receiving sample tubes of a biological reaction
`mixture. As shown in FIGS. 3-5, the thermal system base 15
`includes a thermal block assembly 20. Thermal block assem­
`bly 20 includes a fiat thermal block plate 22 and a plurality
`of sample wells 24 for receiving tubes with samples of
`DNA, as best shown in FIGS. 4, 6 and 7. Thermal block
`
`arranged to form a grid which is sixteen by twenty-four,
`thereby accommodating 384 sample wells. The sample wells
`24 are conical in shape, as shown in FIG. 8. The walls 25 of
`the tube are conical, and extend at an angle to the fiat plate
`20 22. The bottom 26 of the interior of the sample well is
`rounded. The bottom of each sample well 24 is attached to
`the thermal block plate 22. It should be understood that the
`sample wells 24 could have any shape (i.e., cylindrical,
`square or similar shapes), so that the inner surface of the
`25 sample wells 24 closely mates with the sample tube 140
`inserted inside.
`The sample wells 24 are designed so that sample tubes
`140 with DNA samples can be placed in the sample wells 24.
`FIG. 5 shows a partial cut-away cross section with sample
`30 tubes 140 placed in the sample wells 24. Each sample well
`24 is sized to fit the sample tube 140 exterior so that there
`will be substantial contact area between the sample tube 140
`and the interior portion of a sample well wall 25 to enhance
`the heat transfer to the DNA sample in the sample tube 140
`35 and reduce differences between the DNA mixture and
`sample well temperatures. The sample tube 140 includes a
`conical wall portion 142 which closely mates with the
`sample well wall 25.
`The sample tubes 140 are available in three common
`40 forms: (1) single tubes; (2) strips of eight tubes which are
`attached to one another; and (3) tube trays with 96 attached
`sample tubes. The present invention is preferably designed
`to be compatible with any of these three designs. The sample
`tubes 140 may be composed of a plastic, preferably molded
`45 polypropylene, however, other suitable materials are accept­
`able. A typical sample tube 140 has a fluid volume capacity
`of approximately 200 µl, however other sizes and configu­
`rations can be envisaged within the spirit and scope of the
`present invention. The fluid volume typically used in an
`50 experiment is substantially less than the 200 µl sample tube
`capacity.
`Although the preferred embodiment uses sample wells,
`other sample holding structures such as slides, partitions,
`beads, channels, reaction chambers, vessels, surfaces, or any
`55 other suitable device for holding a sample can be envisaged.
`Moreover, although the preferred embodiment uses the
`sample holding structure for biological reaction mixtures,
`the samples to be placed in the sample holding structure are
`not limited to biological reaction mixtures. Samples could
`60 include any type of product for which it is desired to heat
`and/or cool, such as cells, tissues, microorganisms or non­
`biological product.
`Alternatively, a thin film of clear or opaque material could
`be attached (to form a seal) to the tops of the sample
`65 containers in place of a series of caps. This type of sample
`container cover can reduce the labor associated with cap
`installation for some users. The flexible heating cover
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`7
`assembly of the present invention works with this type of
`sealed film container cover. Typically, these films are com­
`posed of a thin plastic with a layer of epoxy which can be
`cured using heat, pressure, heat and pressure, or UV light.
`As embodied herein and shown for example in FIG. 5,
`each sample tube 140 also has a corresponding sample tube
`cap 146 for maintaining the biological reaction mixture in
`the sample tube. The caps 146 are typically inserted inside
`a top cylindrical surface 144 of the sample tube 140. The
`caps 146 are relatively clear so that light can be transmitted
`through the cap 146. The sample tube caps 146 may be
`composed of a plastic, preferably molded polypropylene,
`however, other suitable materials are acceptable. Each cap
`146 has an optical window 148 on the top surface of the cap.
`The optical window 148 in the cap 146 is thin, fiat, com­
`posed of plastic, and allows radiation such as excitation light
`to be transmitted to the DNA samples and emitted fluores­
`cent light from the DNA to be transmitted back to an optical
`detection system during cycling.
`A biological probe can be placed in the DNA samples so
`that fluorescent light is transmitted in and emitted out as the
`strands replicate during each cycle. A suitable optical detec­
`tion system can detect the emission of radiation from the
`sample. The detection system can thus measure the amount
`of DNA which has been produced as a function of the
`emitted fluorescent light. Data can be provided from each
`