MRI: Development of Programmable Substrates for Quantitative Investigation of Mechanotransduction using Holographic Optical Tweezer (HOT) Arrays

Yale University, New Haven CT

Investigators

Abstract

This is an award to develop Holographic Optical Tweezers (HOT) into a instrument for quantitative investigation of mechanotransduction in cells. This instrument will be uniquely capable of quantifying the structure and dynamics of the cytoskeleton with fluorescent speckle microscopy (FSM) while biochemical signals and mechanical forces are simultaneously controlled with HOT. A spinning-disc confocal microscope will provide images of the cytoskeleton for FSM, while a programmable closed-loop HOT system will simultaneously manipulate up to about one hundred colloidal particles in 4-D. These particles will be functionalized with a number of different biochemical molecules and will be attached to the cell membrane in any desired pattern. Simultaneous trapping of multiple beads will enable the application of nanonewton level forces to single cells - an increase of up to two orders of magnitude over existing optical methods. The closed-loop control system will be capable of generating large forces and, with the help of novel low-level feature recognition and hologram calculation codes, it will also be capable of controlling and measuring forces at multiple locations in real time - with resolutions as fine as 10 fN at 30Hz. The investigators will work closely with academic and industrial partners to ensure that the proposed technology is widely adopted. This interdisciplinary project will bring together students and post-docs from engineering and biology. Through the course of the proposed research, they will develop a broad set of skills in colloid science, optics, molecular biology and cell biology. This interdisciplinary training will prepare them for innovative careers in nanoscience and nanotechnology at the intersection of the physical and biological sciences. Furthermore, the proposed instrument will be employed in an advanced course in optical microscopy for graduate students in the physical and biological sciences. The investigators will continue to encourage members of underrepresented groups to contribute to their research.