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US 2012/0058460 A1Published application
Slug control during thermal cycling
AssigneeCanon U.S. Life Sciences, Inc.
InventorsJohnathan S. Coursey, Kenton C. Hasson, Sami Kanderian, Gregory H. Owen, Hongye Liang, Scott Corey, Brian Bean
Life of this patent
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Abstract
The present invention, in one aspect, provides methods and systems for controlling slugs using temperature dependent fluorescent dyes. In some embodiments, the present invention uses one or more techniques to enhance the visibility of slugs, enhance a system's ability to differentiate between slugs, and enhance a system's ability to identify the positions of slugs.
Claims64
A method for controlling the position of a slug in a microfluidic device, comprising:
- exciting a fluorescent dye in said slug;
- acquiring an image of the fluorescence of said slug;
- processing data from a region of interest in said image;
- identifying a position of said slug within said region of interest from said processed image data;
- controlling the position of said slug based on the identified position of said slug.
2 more elements
A method for controlling the position of fluid slugs in a microfluidic device, comprising:
- providing a first slug in said microfluidic device, wherein said first slug comprises a first dye;
- providing a second slug in said microfluidic device, wherein said second slug comprises a second dye;
- acquiring an image of said first and second slugs in said microfluidic device, processing data from a region of interest in said image;
- identifying a boundary between said first slug and said second slug within said region of interest from said processed image data;
- controlling the position of said first slug and said second slug based on the identified boundary between said slugs.
2 more elements
A method for controlling the position of a slug in a microfluidic device, comprising:
- acquiring an image of a slug in a microfluidic device;
- processing data from a region of interest in said image;
- identifying a position of said slug within said region of interest from said processed image data;
- controlling the position of said slug based on the identified position of said slug.
1 more element
A system for controlling the position of a slug within a microfluidic device, comprising:
- (a) a microfluidic device comprising one or more fluid channels and one or more slugs in said one or more fluid channels, wherein said one or more slugs include one or more fluorescent dyes, and
- (b) a slug excitation device configured to excite said one or more fluorescent dyes within said one or more slugs;
- (c) a light-sensing unit configured to capture fluorescence from at least a region of interest of the microfluidic device, wherein said region of interest includes one or more of said slugs;
- (d) a processing unit configured to receive information from said light-sensing unit;
- (e) a position identification unit configured to identify the position of one of said one or more slugs within said region of interest;
- and
- (f) a slug control unit, configured to position the slug within the microfluidic device based upon the identified position.
4 more elements
A method for controlling the position of a sample slug in a microfluidic device, comprising:
- providing a first blanking slug comprising a fluorescent dye and a second blanking slug comprising a fluorescent dye, wherein said first and second blanking slugs surround said sample slug;
- exciting a fluorescent dye in said blanking slugs;
- acquiring an image of said blanking slugs and said sample slug, wherein the image captures the fluorescence of the blanking slugs;
- processing data from a region of interest in said image;
- identifying a position of said sample slug within said region of interest from said processed image data;
- controlling the position of said sample slug based on the identified position of said sample slug.
3 more elements
A method for controlling the position of a sample slug in a microfluidic device, comprising:
- providing a first blanking slug comprising a fluorescent dye and a sample slug, wherein said blanking slug has an interface with said sample slug;
- exciting a fluorescent dye in said blanking slug;
- acquiring an image of at least a portion of said blanking slug and at least a portion of said sample slug;
- processing data from a region of interest in said image;
- identifying a position of said sample slug within said region of interest from said processed image data;
- controlling the position of said sample slug based on the identified position of said sample slug.
3 more elements
A method for controlling the position of a fluid slug during polymerase chain reaction (PCR) amplification, wherein the PCR has a denaturation step, an annealing step and an extension step, comprising:
- exciting a fluorescent dye in said slug;
- acquiring images of the fluorescence of said slug;
- processing data in said images;
- identifying the position of said slug from said processed image data;
- controlling the position of said slug based on the detected position;
- wherein the denaturation step is of shorter duration than the period at which said images are acquired.
3 more elements
A method for controlling the position of a slug during a polymerase chain reaction (PCR) thermal cycle, comprising:
- a denaturation step comprising subjecting the slug to a temperature sufficiently high to allow denaturation,
- an annealing/extension step comprising subjecting the slug to a temperature lower than the denaturation temperature and which is sufficient to allow primer annealing and extension,
- exciting a fluorescent dye in said slug during said thermal cycle;
- acquiring images of the fluorescence of said slug;
- processing data in said images;
- identifying the position of said slug from said processed image data;
- and
- controlling the position of said slug based on the detected position.
5 more elements
A system for controlling the position of a slug within a microfluidic device, comprising:
- (a) a microfluidic device comprising one or more fluid channels, one or more thermal elements in communication with said one or more channels, wherein said one or more fluid channels are configured to receive one or more slugs and wherein said one or more slugs include one or more fluorescent dyes,
- (b) a slug excitation device configured to excite said one or more fluorescent dyes within said one or more slugs;
- (c) a light-sensing unit configured to capture fluorescence at an acquisition rate from at least a region of interest of the microfluidic device, wherein said region of interest is configured to include one or more of said slugs;
- (d) a temperature control unit configured to control the temperature of said slugs using said thermal elements of the microfluidic device, wherein said temperature control unit is configured to subject said slug to (i) a denaturation step at a first temperature for a first duration of time, (ii) an annealing step at a second temperature for a second duration of time, and (iii) an extension step at a third temperature for a third duration of time;
- (e) a processing unit configured to receive information from said light-sensing unit;
- (f) a position identification unit configured to identify the position of one of said one or more slugs within said region of interest;
- and
- (g) a slug control unit, configured to position the slug within the microfluidic device based upon the identified position, and
- wherein the duration of the denaturation step is of a shorter duration than the period between image acquisitions.
6 more elements
A method for controlling the position of a fluid slug during polymerase chain reaction (PCR) amplification, wherein the PCR has a denaturation step, an annealing step and an extension step, comprising:
- exciting a fluorescent dye in said slug;
- acquiring images of the fluorescence of said slug;
- processing data in said images;
- identifying the position of said slug;
- controlling the position of said slug based on the detected position;
- wherein the annealing step is of longer duration than the denaturation period, such that more than 30% of the images collected are obtained during the annealing step.
3 more elements
A method for controlling the position of a fluid slug during polymerase chain reaction (PCR) amplification, wherein the PCR has a denaturation step, an annealing step and an extension step, comprising:
- exciting a fluorescent dye in said slug;
- acquiring images of the fluorescence of said slug;
- processing data in said images;
- identifying the position of said slug;
- controlling the position of said slug based on the detected position;
- wherein the extension step is of longer duration than the denaturation period, such that more than 30% of the images collected are obtained during the extension step.
3 more elements
A method for controlling the position of a fluid slug during a polymerase chain reaction (PCR) amplification,
- wherein the PCR comprises:
- a first denaturation step comprising subjecting the slug to a temperature sufficiently high to allow denaturation, and
- a second annealing and extension step comprising subjecting the slug to a temperature lower than the denaturation temperature to allow primer annealing and extension,
- comprising:
- exciting a fluorescent dye in said slug;
- acquiring images of the fluorescence of said slug;
- processing data in said images;
- identifying the position of said slug;
- and
- controlling the position of said slug based on the detected position.
7 more elements
A system for controlling the position of a slug within a microfluidic device, comprising:
- (a) a microfluidic device comprising one or more fluid channels, one or more thermal elements in communication with said one or more channels, wherein said one or more fluid channels are configured to receive one or more slugs and wherein said one or more slugs include one or more fluorescent dyes,
- (b) a slug excitation device configured to excite said one or more fluorescent dyes within said one or more slugs;
- (c) a light-sensing unit configured to capture fluorescence at an acquisition rate from at least a region of interest of the microfluidic device, wherein said region of interest is configured to include one or more of said slugs;
- (d) a temperature control unit configured to control the temperature of said slugs using said thermal elements of the microfluidic device, wherein said temperature control unit is configured to subject said slug to (i) a denaturation step at a first temperature for a first duration of time, and (ii) an annealing/extension step at a second temperature for a second duration of time.
- (e) a processing unit configured to receive information from said light-sensing unit;
- (f) a position identification unit configured to identify the position of one of said one or more slugs within said region of interest;
- and
- (g) a slug control unit, configured to position the slug within the microfluidic device based upon the identified position.
5 more elements
Dependencies are inferred from the claim text.