Abstract
We use a spatial light modulator (SLM) to diffract a single UV laser pulse to ablate multiple points on a Drosophila embryo. This system dynamically generates a phase hologram for ablating a user-defined pattern fast enough to be used with living, and thus moving, tissue. We demonstrate the ability of this single-pulse multi-point system to perform two experiments that are very difficult for conventional microsurgery—isolating single cells in vivo and measuring fast retractions from large incisions.
Highlights
Laser microsurgery is a well-established method for studying the cellular forces that drive morphogenesis [1,2,3,4,5]
The first pulse ablates unaltered tissue; all subsequent pulses ablate tissue that is already retracting or undergoing strain relaxation. This motion makes some potentially useful microsurgeries very challenging—e.g., cutting around a single cell or patch of tissue to mechanically isolate it from a surrounding epithelium—and strongly interferes with the quantitative measurement of retraction velocity after extended incisions [1]
We show how nanosecond UV lasers can be coupled with an spatial light modulator (SLM) to provide new and innovative means to probe cellular mechanics
Summary
Laser microsurgery is a well-established method for studying the cellular forces that drive morphogenesis [1,2,3,4,5]. Microsurgery creates extended incisions in a tissue by ablating discrete points one at a time—a serial multi-pulse procedure. Such incisions have proven useful in determining the relative morphogenetic roles of different tissue regions; the quantitative interpretation of such experiments is limited. The first pulse ablates unaltered tissue; all subsequent pulses ablate tissue that is already retracting or undergoing strain relaxation This motion makes some potentially useful microsurgeries very challenging—e.g., cutting around a single cell or patch of tissue to mechanically isolate it from a surrounding epithelium—and strongly interferes with the quantitative measurement of retraction velocity after extended incisions [1]. We do so by dynamically shaping the phase profile of a single laser pulse using a programmable spatial light modulator (SLM)
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