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A method of microperfusion with oxygenated saline as applied to an insect brain


Hengstenberg,  R
Department Human Perception, Cognition and Action, Max Planck Institute for Biological Cybernetics, Max Planck Society;

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Hengstenberg, R. (1982). A method of microperfusion with oxygenated saline as applied to an insect brain. Journal of Neuroscience Methods, 6(1-2), 169-171. doi:10.1016/0165-0270(82)90027-9.

For electrophysiological studies in the fly's optic lobe, a procedure was required to perfuse the brain at controllable rates with oxygenated solutions, while recording intracellularly from visual interneurons. The cornea of the compound eyes has to be left in air, in order not to disturb their optics, and to allow physiological stimulation. When the head capsule is opened to get acces to the brain, air sacs and major tracheal trunks tend to collapse by either of two causes: surface tension may flatten tracheae at too low fluid levels and hydrostatic pressure may flatten tracheae at too high levels. Only within a small range of saline levels ( < ± 30µm ) main tracheae and air sacs were found to remain inflated in the dissected head. Therefore the saline level had to be precisely controlled and stabilized within a few microns. With this procedure, it is possible to maintain the fly's brain (< 5 µl ) alive for more than 24 hrs in a perfused volume of only 15 µl, and to record intracellularly from fibres as small as 2 µm. The same procedure should be applicable wherever small, open volumes have to be precisely perfused, and mechanical or electrical artifacts cannot be tolerated.