Amnis Corporation

Richard Bauer

William E Ortyn

David Abraham Basiji

Keith Frost

David Joseph Perry

Brian E Hall

James K Riley

Luchuan Liang

Rosalynde J Finch

David Perry

Rosalynde Finch

A high speed, high-resolution flow imaging system is modified to achieve extended depth of field imaging. An optical distortion element is introduced into the flow imaging system. Light from an object, such as a cell, is distorted by the distortion element, such that a point spread function (PSF) of the imaging system is invariant across an extended depth of field. The distorted light is spectrally dispersed, and the dispersed light is used to simultaneously generate a plurality of images. The images are detected, and image processing is used to enhance the detected images by compensating for the distortion, to achieve extended depth of field images of the object. The post image processing preferably involves de-convolution, and requires knowledge of the PSF of the imaging system, as modified by the optical distortion element.

Extended depth of field imaging for high speed object analysis

Combinatorially-synthesized deoxyribonucleic acid (DNA) oligonucleotides attached to encoded beads that are hybridized to amplified and labeled genomic DNA or ribonucleic acid (RNA) are analyzed using a flow imaging system. Oligonucleotides and corresponding reporters are bound to the surfaces of a plurality of small beads such that different beads bear different oligo sequences. Each bead bears a unique optical signature comprising a predefined number of unique reporters, where each reporter comprises a predefined combination of different fluorochromes. The composite spectral signature in turn identifies the unique nucleotide sequence of its attached oligo chains. This optical signature is rapidly decoded using an imaging system to discriminate the different reporters attached to each bead in a flow in regard to color and spatial position on the bead.

Method and apparatus for reading reporter labeled beads

Frequency domain velocity measurements and time domain velocity measurements are made using light from cells or other objects. An optical grating is used to modulate the light from an object so that it has a frequency proportional to the velocity of the object. Depending upon the embodiment, the pitch of the optical grating is uniform or varying. The modulated light is detected and various signal processing techniques, such as a Fast Fourier Transform function, are used to indicate the velocity of the object. Preferably, the velocity measured is applied in determining a timing signal employed for synchronization of an image of the object and an detector signal in an optical analysis system that uses a time delay integration detector to determine characteristics of the object in response to light from the object.

Measuring the velocity of small moving objects such as cells

Frequency domain velocity measurements and time domain velocity measurements are made using light from cells or other objects. An optical grating is used to modulate the light from an object so that it has a frequency proportional to the velocity of the object. Depending upon the embodiment, the pitch of the optical grating is uniform or varying. The modulated light is detected, producing an analog signal that is then digitally sampled. Time domain signal processing techniques are used to determine the velocity of the object from the digital samples. Preferably, the velocity measured is applied in determining a timing signal employed for synchronization of an image of the object and an detector signal in an optical analysis system that uses a time delay integration (TDI) detector to determine characteristics of the object in response to light from the object.

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Kirkland, WA, United States of America

Richard Bauer