Abstract
The efficacy of funnel-freezing of rat brain to inactivate metabolic processes and preserve in vivo tissue glucose concentration was validated by comparing the results obtained by funnel-freezing with those obtained with freeze-blowing of brain. The arterial plasma glucose level was clamped at 9 mM in halothane-anesthetized rats to produce identical glucose levels in brain tissue prior to freeze fixation. In funnel-frozen and freeze-blown brains, tissue glucose concentrations were 2.47 +/- 0.05 and 2.47 +/- 0.06 mumol/g (means +/- SEM), respectively. Lactate levels in funnel-frozen brains were slightly but significantly higher than those in freeze-blown brains, i.e., 1.56 +/- 0.05 mumol/g versus 1.30 +/- 0.05 mumol/g (means +/- SEM; p less than 0.05). Regional analysis in funnel-frozen brains revealed that glucose concentrations in superficial and basal brain areas remained approximately equal at 2.30 +/- 0.1 mumol/g and 2.31 +/- 0.09 mumol/g (means +/- SEM), respectively. Our findings indicate that in the anesthetized rat, funnel-freezing of brain is suitable for the measurement of regional in vivo glucose concentrations.
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