Assembly system for in situ per amplification

• US 5,675,700 A
• Filed: 06/05/1995
• Issued: 10/07/1997
• Est. Priority Date: 02/16/1993
• Status: Expired due to Term

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: The Perkin-Elmer Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Haff, Lawrence A., Atwood, John G.
• Primary Examiner(s): HOANG, TU BA
• Application Number:
  US08/462,926
• Time in Patent Office: 855 Days
• Field of Search: 392/382, 392/379, 219/541, 219/385, 219/200, 219/201, 219/521, 435/288.3-288.5, 435/809, 435/287.2, 422/99, 422/102, 422/104, 359/395, 359/391, 359/392, 359/396-398, 935/85-87, 34/202, 356/244, 211/41
• US Class Current: 392/382
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2300/041 Connecting closures to device or container
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 3/508 rigid containers not provided for above
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 9/52 Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
• CPC Class Codes: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2543/101 in situ amplification
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers

Abstract

A complete in situ PCR system for amplification of nucleic acids contained in a prepared cell or tissue sample. The containment system for the PCR sample comprises a glass microscope slide, a specimen sample containing the target nucleic acid sequence mounted on the slide, a flexible plastic cover over the sample, and a retaining assembly fastened to the slide and to the cover to retain and seal a reaction mixture against the sample during thermal cycling.

The retaining assembly includes a rigid ring on a rim portion of the cover, a cross beam having spaced parallel rails joined by opposite flat ends, and a pair of clips which are pressed over the ends and opposite sides of the slide to fasten the cross beam and cover to the slide.

Claims

1. An apparatus for assembly of an in situ PCR sample and reagent fluid containment system on a glass slide wherein said containment system comprises a cover member and a retaining assembly, said apparatus comprising:

a base;

a support platform mounted on said base, said support platform having a generally horizontal top surface for receiving and supporting components of said retaining assembly and said slide in a pre-determined spaced relation;

an elongate compression arm movably supported from said base at a position spaced from said platform, said arm being operative to compress said glass slide and said retaining assembly together in close contact; and

a retaining assembly installation mechanism positioned so as to install said retaining assembly onto said slide while said compression arm is compressing said glass slide and said retaining assembly together.

2. The apparatus of claim 1, wherein said mechanism comprises a pair of pliers having finger portions positioned adjacent opposite edges of said horizontal top surface of said support platform and moving portions operative to move said finger portions toward each other.

3. The apparatus of claim 2 wherein each of said pliers has a handle portion coupled to one of said finger portions.

4. The apparatus according to claim 1 further comprising a heating means operatively coupled to said apparatus for heating components of said retaining assembly and said slide.