Light transmittance as an index of cell volume in hippocampal slices: optical differences of interfaced and submerged positions

Light transmittance (T) in the CA1 region of hippocampal slices was measured during exposure to media of various osmolarities to determine the utility of optical measurements as an index of changes in cell volume. In slices positioned at the gas-liquid interface, hypo-osmotic medium consistently produced a decrease in T and hyperosmotic medium produced an increase in T. The magnitude of deltaT was graded as a function of the strength of osmotic change. All changes in T were reversible upon return to isosmotic medium. In contrast, osmotically induced changes in T in submerged slices were consistently opposite in direction to those observed in slices at the interface. The magnitude and direction of deltaT could be altered by systematic variation of the level of the bathing medium within the same chamber, indicating that both extrinsic optical properties of various interfaces, such as refraction and reflection, and intrinsic optical properties of the tissue contribute to the observed T. Spectral measurements eliminated the possibility that osmotically induced deltaT was the result of changes in light absorbance by intrinsic chromophores such as cytochromes or hemoglobin. The results show that measurements of deltaT can be a useful index of changes in cell volume in brain slices, provided that the level of the bath remains constant.