Immunohistochemical and electronmicroscopic effects of a new 2.1 microns Ho:YAG laser on the rat brain

Using an experimental animal model, the thermal single-pulse lesion derived from a mid-infrared 1.0 Joule 300 microns fibre-conducted Holmium: Yttrium-Aluminum-Garnet (Ho:YAG) laser was examined, with special emphasis on the orientation and depth of the tissue reaction. Performing biparietal craniotomy in Sprague-Dawley rats weighing 250-300 g, both hemispheres were targeted by different radiant exposures from 20 to 140 J/cm2 derived from a 600-800 microsecond single pulse. After survival periods of one to 30 days, the animals were sacrificed and both hemispheres were processed for light- and electronmicroscopic investigations. To resolve the depth and orientation of the tissue reaction regarding the localization of reactive astrocytes, we looked for the expression of glial proteins like glial fibrillary acidic protein (GFAP), Vimentin and S 100 with a three-step biotin-avidin immunoperoxidase method. Neuronal and secondary axonal damage was investigated by labelling Neurofilament and Synaptophysin. The tissue reaction beneath the ablated material, consisting of a vacuolation and coagulation zone resulting from heat diffusion, was further elucidated by localization of the heat shock protein (HSP 72 kilo Dalton). Revealing the extension of reactive astrocytes and the degree of the electronmicroscopically depicted glial oedema, the depth of the tissue damage was estimated to reach about 700 microns beneath laser excision. Since McKenzie predicted the depth of tissue damage beneath CO2 and YAG laser excisions in a theoretical mathematical model, the authors were able to develop a sensitive model for testing new laser systems and as a promising instrument for neurosurgery.