Abstract
Surgical procedures as a prelude to optical imaging are a rate-limiting step in experimental neuroscience. Towards automation of these procedures, we describe the use of nonlinear optical techniques to create a thinned skull window for transcranial imaging. Metrology by second harmonic generation was used to map the surfaces of the skull and define a cutting path. Plasma-mediated laser ablation was utilized to cut bone. Mice prepared with these techniques were used to image subsurface cortical vasculature and blood flow. The viability of the brain tissue was confirmed via histological analysis and supports the utility of solely optical techniques for osteotomy and potentially other surgical procedures.
Highlights
Optical imaging techniques provide a powerful and increasingly popular means to probe neuronal function in the mammalian central nervous system
The second harmonic light is generated in the forward direction, yet back-scattering by the bone and underlying soft tissue allows the second harmonic generation (SHG) signal to be detected by the incident beam optics and makes SHG a suitable technique for profiling the skull
We ask if an automated surgical procedure, which avoids the use of mechanical tools but rather utilizes SHG for metrology and plasma-mediated laser ablation for the controlled removal of bone, is a viable means for osteotomy [26]
Summary
Optical imaging techniques provide a powerful and increasingly popular means to probe neuronal function in the mammalian central nervous system. The second harmonic light is generated in the forward direction, yet back-scattering by the bone and underlying soft tissue allows the SHG signal to be detected by the incident beam optics and makes SHG a suitable technique for profiling the skull. Plasma-mediated laser ablation via ultrashort pulsed laser light [14,15,16,17,18,19] provides a way to remove material in highly localized volumes at the focus of the laser beam, a volume on the order of 0.1 to 10 μm3 [20]. We ask if an automated surgical procedure, which avoids the use of mechanical tools but rather utilizes SHG for metrology and plasma-mediated laser ablation for the controlled removal of bone, is a viable means for osteotomy [26]. Our test bed is the fabrication of a thinned skull transcranial window in mice [3], with the goal of utilizing such windows for in vivo imaging of labeled brain vasculature and functional imaging of the underlying blood flow
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