title: Brain tissue stiffness is a sensitive marker for acidosis.
abstract: Carbon dioxide overdose is frequently used to cull rodents for tissue harvesting. However, this treatment may lead to respiratory acidosis, which potentially could change the properties of the investigated tissue.
  
  
  
  Mechanical tissue properties often change in pathological conditions and may thus offer a sensitive generic readout for changes in biological tissues with clinical relevance. In this study, we performed force-indentation measurements with an atomic force microscope on acute cerebellar slices from adult rats to test if brain tissue undergoes changes following overexposure to CO2 compared to other methods of euthanasia.
  
  
  
  The pH significantly decreased in brain tissue of animals exposed to CO2. Concomitant with the drop in pH, cerebellar grey matter significantly stiffened. Tissue stiffening was reproduced by incubation of acute cerebellar slices in acidic medium.
  
  
  
  Tissue stiffness provides an early, generic indicator for pathophysiological changes in the CNS. Atomic force microscopy offers unprecedented high spatial resolution to detect such changes.
  
  
  
  Our results indicate that the stiffness particularly of grey matter strongly correlates with changes of the pH in the cerebellum. Furthermore, the method of tissue harvesting and preparation may not only change tissue stiffness but very likely also other physiologically relevant parameters, highlighting the importance of appropriate sample preparation.
authors.affiliation: Department of Physics, University of Cambridge, J J Thomson Avenue, Cambridge CB3 0HE, United Kingdom.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Holtzmann
authors.affiliation: Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3DY, United Kingdom; Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, United Kingdom.
authors.forename: Hélène O B
authors.initials: HO
authors.lastname: Gautier
authors.affiliation: Department of Physics, University of Cambridge, J J Thomson Avenue, Cambridge CB3 0HE, United Kingdom.
authors.forename: Andreas F
authors.initials: AF
authors.lastname: Christ
authors.affiliation: Department of Physics, University of Cambridge, J J Thomson Avenue, Cambridge CB3 0HE, United Kingdom; Biotechnology Center, Technische Universität Dresden, Tatzberg 47/49, 01307 Dresden, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Guck
authors.affiliation: Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, United Kingdom.
authors.forename: Ragnhildur Thóra
authors.initials: RT
authors.lastname: Káradóttir
authors.affiliation: Department of Physics, University of Cambridge, J J Thomson Avenue, Cambridge CB3 0HE, United Kingdom; Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3DY, United Kingdom. Electronic address: kf284@cam.ac.uk.
authors.forename: Kristian
authors.initials: K
authors.lastname: Franze
mesh_terms: D000138:Acidosis / Q000175:diagnosis* / Q000503:physiopathology*; D000818:Animals; D001696:Biomechanical Phenomena; D001921:Brain / Q000503:physiopathology*; D002245:Carbon Dioxide / Q000633:toxicity; D004548:Elasticity; D006863:Hydrogen-Ion Concentration; D018625:Microscopy, Atomic Force / Q000379:methods; D051381:Rats; D046509:Tissue Culture Techniques
chemical_list: D002245:Carbon Dioxide
keywords: AFM; Acidosis; Carbon dioxide; Cerebellum; Mechanical properties; Tissue mechanics