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US 11,112,591 B2Granted
Reduced dimensionality structured illumination microscopy with patterned arrays of nanowells
AssigneesIllumina, Inc., Illumina Cambridge Limited
InventorsGary Mark Skinner, Geraint Wyn Evans, Stanley S. Hong
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Abstract
Techniques are described for reducing the number of angles needed in structured illumination imaging of biological samples through the use of patterned flowcells, where nanowells of the patterned flowcells are arranged in, e.g., a square array, or an asymmetrical array. Accordingly, the number of images needed to resolve details of the biological samples is reduced. Techniques are also described for combining structured illumination imaging with line scanning using the patterned flowcells.
Claims18
A method of imaging a biological sample, comprising:
- projecting an optical pattern onto a biological sample and capturing a first image of the optical pattern overlaid on the biological sample, the biological sample being contained in an asymmetrically patterned flowcell comprising a plurality of elongated nanowells, wherein each of the elongated nanowells is elliptically shaped or rectangularly shaped;
- phase shifting the projected optical pattern relative to the biological sample and capturing at least a second image of the phase shifted optical pattern overlaid on the biological sample;
- and
- reconstructing a high resolution image representative of the biological sample based upon the first captured image and the at least second captured image.
1 more element
A method of imaging a biological sample, comprising:
- directing light through an optical diffraction grating in a first phase and angle orientation;
- projecting an optical diffraction grating pattern generated by the light being directed through the optical diffraction grating onto the biological sample and capturing a first image of the optical diffraction grating pattern overlaid on the biological sample, the biological sample being contained in an asymmetrically patterned flowcell comprising a plurality of elongated nanowells, wherein each of the elongated nanowells is elliptically shaped or rectangularly shaped;
- phase shifting the optical diffraction grating, projecting the optical diffraction grating pattern onto the biological sample and capturing at least a second image of the phase shifted optical diffraction grating pattern overlaid on the biological sample;
- reorienting the optical diffraction grating to a second angle orientation, projecting the optical diffraction grating pattern onto the biological sample, and capturing a third image of the optical diffraction grating pattern overlaid on the biological sample;
- phase shifting the optical diffraction grating, projecting the optical diffraction grating pattern onto the biological sample and capturing at least a fourth image of the phase shifted optical diffraction grating pattern overlaid on the biological sample;
- and
- reconstructing a high resolution image representative of the biological sample based upon the first, the at least second, the third, and the at least fourth captured images.
4 more elements
A system, comprising:
- a laser source emitting a light beam;
- an optical diffraction grating adapted to generate an optical diffraction grating pattern upon passage of the emitted light beam through the optical diffraction grating;
- a camera assembly adapted to capture a plurality of images of optical diffraction grating pattern overlaid on a biological sample, the plurality of images reflecting three phases of the optical diffraction grating relative to the biological sample, wherein the biological sample is located in a flowcell comprising a plurality of nanowells that are oriented in an asymmetrical array and wherein each of the plurality of nanowells are elliptically shaped or rectangularly shaped;
- and
- a processor adapted to reconstruct a high resolution image representative of the biological sample based on a combination of the plurality of images.
2 more elements
A system, comprising:
- a laser source emitting a light beam;
- an optical diffraction grating adapted to generate an optical diffraction grating pattern upon passage of the emitted light beam through the optical diffraction grating;
- a camera assembly adapted to capture a plurality of images of optical diffraction grating pattern overlaid on a biological sample, the plurality of images reflecting three phases of the optical diffraction grating relative to the biological sample, wherein the camera assembly comprises a time delay integration line scanning camera assembly;
- and
- a processor adapted to reconstruct a high resolution image representative of the biological sample based on a combination of the plurality of images.
2 more elements
A system, comprising:
- a laser source emitting a light beam;
- an optical diffraction grating adapted to generate an optical diffraction grating pattern upon passage of the emitted light beam through the optical diffraction grating;
- a camera assembly adapted to capture a plurality of images of optical diffraction grating pattern overlaid on a biological sample, the plurality of images reflecting three phases of the optical diffraction grating relative to the biological sample and two angular orientations of the optical diffraction grating relative to the biological sample, wherein the biological sample is located in a flowcell comprising a plurality of nanowells that are oriented in an asymmetrical array and wherein each of the plurality of nanowells are elliptically shaped or rectangularly shaped;
- and
- a processor adapted to reconstruct a high resolution image representative of the biological sample based on a combination of the plurality of images.
2 more elements
Dependencies are inferred from the claim text.