Methods and compositions for retrieving cellular structures based on spatiotemporal profiles

Inventors

St-Pierre, François • LEE, Jihwan • Liu, Zhuohe • Suzuki, Peter

Assignees

Baylor College of Medicine • William Marsh Rice University

Abstract

Provided herein are embodiments of methods and systems for screening cellular, subcellular, and multicellular structures. In one embodiment, a method for screening is provided comprising the steps of introducing a plurality of cellular, subcellular, or multicellular structures, or a combination thereof, to an imaging system, wherein one or more structures of the plurality comprise one or more taggable markers; imaging the plurality of structures using the imaging system; identifying one or more target structures among the plurality of structures based on one or more properties of the target structures; tagging the target structures to produce tagged target structures, wherein each target structure is selectively illuminated by an excitation light, thereby causing one or more taggable markers within the target structure to be phototransformed to produce one or more phototransformed taggable markers within the target structure; and isolating one or more tagged target structures from the plurality of structures.

Core Innovation

The invention provides an all-optical, microscopy-based screening platform for identifying target cellular, subcellular, or multicellular structures. A plurality of structures is introduced to an imaging system, and one or more structures of the plurality comprise one or more taggable markers. The plurality is imaged, and one or more target structures are identified based on one or more properties of the target structures.

The identified target structures are then selectively tagged by selectively illuminating each target structure with excitation light. This causes one or more taggable markers within the target structure to be phototransformed to produce one or more phototransformed taggable markers within the target structure. The phototransformed taggable markers are used to drive downstream recovery of the tagged targets.

After phototransformation, one or more tagged target structures are isolated from the plurality of structures using the phototransformed taggable markers. Where the plurality further comprises one or more selectable markers, the phototransformed taggable markers within the target structure cause activation of one or more selectable markers. Isolation is performed by culturing the plurality in an environment only amenable to structures comprising the one or more activated selectable markers to produce one or more isolated tagged target structures.

Claims Coverage

The document provides one independent claim covering an imaging-based screening workflow with selective phototransformation tagging and isolation. The claim includes inventive features spanning imaging and property-based target identification, selective phototransformation of taggable markers in illuminated target structures, and culture-based isolation enabled by activated selectable markers driven by the phototransformed taggable markers.

Across the independent claim, the inventive coverage centers on imaging-based screening with property-driven target identification, selective excitation-driven phototransformation tagging, and isolation of tagged structures using phototransformed taggable markers, optionally coupled to activation and culture-only selection based on selectable markers.

Stated Advantages

Documented Applications

No documented applications found