Sub-Surface, Micrometer-Scale Incisions Produced in Rodent Cortex using Tightly-Focused Femtosecond Laser Pulses

Background and Objective: Techniques that allow targeted, micrometer-scale disruption in the depths of biological tissue, without affecting overlying structures or causing significant collateral damage, could potentially lead to new surgical procedures. We describe an optical technique to make sub-surface incisions in in vivo rodent brain and characterize the relationship between the cut width and maximum depth of these optical transections as a function of laser energy. Materials and Methods: To produce cuts, high intensity, femtosecond laser pulses were tightly focused into and translated within the cortex, through a craniotomy, in anesthetized rodents. Imaging of stained brain slices was used to characterize cut width and maximum cutting depth. Results: Cut width decreased exponentially as a function of depth and increased as the cube root of laser energy, but showed about 50% variation at fixed depth and laser energy. For example, at a laser energy of 13 mu J, cut width decreased from 158 +/- 43.1 mu m (mean +/- standard deviation) to 56 +/- 33 mu m over depths of approximately 200-800 mu m, respectively. Maximal cut depth increased logarithmically with laser energy, with cut depths of up to 1 mm achieved with 13 mu J pulses. We further show-cased this technique by selectively cutting sub-surface cortical dendrites in a live, anesthetized transgenic mouse. Conclusions: Femtosecond laser pulses provide the novel capacity for precise, sub-surface, cellular-scale cuts for surgical applications in optically scattering tissues. Lasers Surg. Med. 43:382-391, 2011. (C) 2011 Wiley-Liss, Inc.