Capillary array electrophoresis scanner
AssigneeAffymetrix, Inc.
InventorsRichard A. Mathies, James R. Scherer
Life of this patent
Estimating the term…
Abstract
An improved rotary confocal fluorescence scanner capable of detecting analytes separated on over a 1,000 capillaries simultaneously. This system uses a confocal microscope objective and mirror assembly that rotates inside a vertical ring of capillaries to provide rapid and efficient excitation and detection of fluorescently labeled fragments separated within a cylindrical capillary array. Use of automated procedures to load and run all capillaries permits one to read more than 350,000 base pairs of raw sequence data per hour.
Claims30
A rotary capillary array scanner, the scanner comprising:
- a source of radiant energy of a first wavelength;
- one or more cylinders having a common vertical axis;
- a plurality of capillaries disposed along the circumferences of the one or more cylinders such that a length of the capillaries is substantially parallel to the vertical axis;
- an objective lens housed within the one or more cylinders, capable of focusing radiant energy of the first wavelength onto the capillaries and capable of receiving radiant energy of a second wavelength emitted from the capillaries;
- a rotor which rotates the objective lens along the circumference of the one or more cylinders;
- a beam splitter capable of passing radiant energy of the second wavelength and rejecting radiant energy of the first wavelength;
- a detector capable of measuring the intensity of the radiant energy passed from the beam splitter.
A rotary capillary array scanner, the scanner comprising:
- a source of radiant energy of a given wavelength;
- one or more cylinders having a common vertical axis;
- a plurality of capillaries disposed along the circumferences of the one or more cylinders such that a length of the capillaries is substantially parallel to the vertical axis;
- an objective lens housed within the one or more cylinders, capable of focusing radiant energy of the given wavelength onto the capillaries and capable of receiving radiant energy of different wavelengths reflected from the capillaries;
- a rotor which rotates the objective lens along the circumference of the one or more cylinders;
- a plurality of beam splitters, each capable of passing the radiant energy emitted from the capillaries at wavelengths different from other beam splitters in the scanner and capable of rejecting all other wavelengths;
- a plurality of photodetectors, each capable of measuring the intensity of radiant energy passed to it by one of the beam splitters.
A method for detecting fluorescence from organic analytes electrophoretically separated in a plurality of capillaries, the method comprising the steps of
- positioning the capillaries around the circumference of at least one cylinder;
- providing a source of radiant energy of a first wavelength,
- focusing, through an objective lens, the radiant energy of the first wavelength onto a capillary so that the radiant energy illuminates analytes within the capillary;
- receiving, through the objective lens, radiant energy of a second wavelength from the analytes;
- passing radiant energy of the second wavelength from the objective lens through a dichromic beamsplitter, rejecting radiant energy of all other wavelengths thereby;
- passing radiant energy of the second wavelength from the dichroic beamsplitter to a photodetector;
- measuring the intensity of radiant energy of the second wavelength with the photodetector;
- rotating the objective lens around the circumference of the at least one cylinder to place another capillary within the path of radiant energy of the first wavelength;
- repeating the steps of focusing, receiving, and measuring the radiant energy and rotating the objective lens until all desired capillaries have been placed within the path of the radiant energy.
Dependencies are inferred from the claim text.