# US 7,745,205 B2 · Container for carrying out and monitoring biological processes > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,745,205 B2 - Document ID: US7745205B2 - Status: Granted - Kind: B2 - Application number: 10/914,648 - Filed: 2004-08-09 - Published: 2005-03-24 - Granted: 2010-06-29 - Earliest claimed: 1996-06-04 - Assignees: University of Utah Research Foundation - Inventors: Carl T. Wittwer; Kirk M. Ririe - Web page: https://patentgrep.com/patent/US7745205B2 ## CPC - G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates} - B01L 3/5082: {Test tubes per se} - B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples} - C12Q 1/6818: involving interaction of two or more labels, e.g. resonant energy transfer - C12Q 1/6823: Release of bound markers - C12Q 1/686: Polymerase chain reaction [PCR] - G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)} - B01L 2300/0838: Capillaries - B01L 2300/1844: using fans - G01N 21/0303: {Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment (G01N21/031 - G01N21/15 take precedence)} - G01N 21/07: Centrifugal type cuvettes (G01N21/09 takes precedence) - G01N 30/02: Column chromatography - G01N 30/06: Preparation - G01N 30/466: {with separation columns in parallel} - G01N 35/025: {having a carousel or turntable for reaction cells or cuvettes} - G01N 2021/6482: {Sample cells, cuvettes} - G01N 2035/00237: {Handling microquantities of analyte, e.g. microvalves, capillary networks} - Y10S 435/808: Optical sensing apparatus - Y10S 435/809: Incubators or racks or holders for culture plates or containers - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS ## Abstract A thermal cycling method and device is disclosed. The device comprises a sample chamber whose temperature can be rapidly and accurately modulated over a range of temperatures needed to carry out a number of biological procedures, such as the DNA polymerase chain reaction. Biological samples are placed in containers each comprising a reservoir and a reaction portion, wherein the reaction portion has a small volume. The small volume reaction portion permits the rapid and accurate temperature modulation. With an optically transmissible reaction portion, DNA amplification may be monitored by fluorescence during PCR. ## Claims ### Claim 1 A system comprising a plurality of containers, said containers comprising walls composed of an optically transparent material and defining a volume having a first and second dimension, wherein the first dimension is less than the second dimension and the ratio of volume to external surface area of the container is less than 1 mm; a continuously rotatable carousel for holding said plurality of containers and sequentially moving the plurality of containers, one-by-one to a monitoring position; a source of forced air for heating and cooling said plurality of containers; and means for performing fluorescent detection on a sample contained within one of said plurality of containers and held at said monitoring position, while each sample is simultaneously subjected to rapid thermal cycling. ### Claim 2 (depends on 1) A system according to claim 1, further comprising a stepper motor and drive shaft for rotating said rotatable carousel, and wherein said fluorescent detection means comprise a fluorimeter assembly for illuminating and detecting fluorescence of a sample held within the container. ### Claim 3 (depends on 2) A system according to claim 2, wherein the containers comprise capillary tubes having a closed end, an open end and an inner diameter of less than 1 mm. ### Claim 4 (depends on 3) A system according to claim 3, wherein the fluorimeter assembly illuminates and detects fluorescence along an axis essentially parallel to the longitudinal axis of the container to establish an epifluorescent excitation and detection pathway. ### Claim 5 (depends on 4) A system according to claim 4, wherein the detection pathway is focused at the closed end tip of the capillary tube. ### Claim 6 (depends on 5) A system according to claim 5, wherein the tip of the capillary tube is positioned at the focal point of the detection pathway during monitoring. ### Claim 7 (depends on 3) The system of claim 3 further comprising a structure that seals the open end of the capillary tubes. ### Claim 8 (depends on 1) A system according to claim 1 wherein the container further comprises a biological sample, wherein the sample comprises nucleic acids, components for amplification of the nucleic acids using a polymerase chain reaction, and a fluorescent probe. ### Claim 9 (depends on 8) A system according to claim 8, wherein the fluorescent probe comprises a fluorophore bound to an oligonucleotide. ### Claim 10 (depends on 9) A system according to claim 9, wherein the fluorescent probe comprises a pair of oligonucleotide probes that hybridize to adjacent regions of said nucleic acids, wherein one of said pair of probes is labeled with an acceptor fluorophore and the other probe is labeled with a donor fluorophore of a fluorescent energy transfer pair. ### Claim 11 A system for conducting and monitoring rapid PCR on multiple samples, said system comprising a plurality of containers, wherein said containers comprise: a capillary tube portion which is closed at one end, open at the other end and comprises an optically transparent portion; and a funnel-shaped portion, having a sample receiving port and a sample transfer port, said funnel-shaped portion being joined to the open end of the capillary tube portion, so that the sample transfer port of the funnel-shaped portion and the open end of the vessel are in alignment, wherein said container holds less than 1 ml of the sample; a removable structure that seals the sample receiving port of the funnel-shaped portion; a continuously rotatable carousel for holding said plurality of containers and sequentially moving the plurality of containers, one-by-one to a monitoring position; and a fluorimeter assembly for illuminating and detecting fluorescence of a sample held within the container at said monitoring position while all samples are being simultaneously subjected to rapid thermal cycling, wherein hot and cool forced air is used as the thermal transfer agent to conduct the rapid thermal cycling. ### Claim 12 (depends on 11) A system according to claim 11, wherein the capillary tubes have an inner diameter of less than 1 mm. ### Claim 13 (depends on 11) The system of claim 11 wherein the capillary tube portion wall is about 0.1 mm thick, and the material comprising the capillary tube portion has a thermal conductivity of about 20 to about 35 in accordance with the formula: ( cal ⁢ ⁢ cm cm 2 ⁢ ⁢ s ⁢ ⁢ degree ⁢ ⁢ C . ) × 10 4 . ### Claim 14 (depends on 13) The system of claim 13, wherein the tip of the capillary tube is positioned at the focal point of the detection pathway during illumination and fluorescence detection of a sample held within the container. ### Claim 15 (depends on 11) The system of claim 11 wherein the structure is a stopper.