Calico Life Sciences LLC

Andrew Gregory York

Sanjay Ramesh Varma

Maria Del Mar Ingaramo

Methods and apparatus for inferring protein binding based on rotational diffusion of a collection of fluorophores. One example of a method includes applying a first light pulse to excite a plurality of fluorophores in the collection of fluorophores to produce a plurality of excited fluorophores, the first light pulse having a first polarization and the plurality of excited fluorophores having a component of their orientation aligned with the first polarization, applying a second light pulse to stimulate emission by the plurality of excited fluorophores, the second light pulse having a second polarization orthogonal to the first polarization, after a time delay following application of the second light pulse, applying a third light pulse of the second polarization to further stimulate emission by the plurality of excited fluorophores, detecting polarized emissions from the plurality of excited fluorophores, and inferring the rate of rotational diffusion of the collection of fluorophores based on the detected polarized emissions.

System and method for inferring protein binding

Fluorescence imaging based on stimulated emission. In one example a system includes an excitation source () that excites fluorescent markers () in a sample () with an excitation beam (), a stimulation source () that stimulates the excited fluorescent markers () to produce stimulated emission (), and an objective lens () that receives the stimulation beam () and the stimulated emission (). The system further includes a phase shifting and attenuation apparatus () configured to receive the stimulation beam () and the stimulated emission () from the objective lens (), to attenuate the stimulation beam (), and to induce a relative phase shift between a reference beam (optionally being at least a portion of the stimulation beam) and the stimulated emission, thereby causing constructive interference between the reference beam and the stimulated emission. The system further includes an optical apparatus () configured to focus the stimulated emission onto an image plane, and a detector () positioned at the image plane and configured to image the stimulated emission.

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San Francisco, CA, United States of America

Andrew Gregory York