The effect of paraformaldehyde fixation and PBS storage on the water content of the human lens

PURPOSE Fixation and phosphate buffered saline (PBS) storage are frequently used before studies of the morphological, biochemical, and optical properties of the human lens begin. It is assumed that this does not alter the properties being examined. The present study was undertaken to determine the effects of fixation and PBS storage on the human lens wet weight. METHODS Human donor lenses were incubated in a buffered paraformaldehyde (PF) solution or in PBS and their wet weights were monitored for up to 44 and 13 days, respectively. RESULTS PF fixation resulted in a large decrease in wet weight, averaging 25%+/-2.3% at 30 days for 14 human donor lenses, aged 49-80 years. The loss was essentially complete by 21 days. Out of the 10 lenses, aged 52-71 years, which were incubated in PBS alone, six of them increased in weight by an average of 38% over 13 days and four ruptured within four days. Comparison of literature data for a fixed eight-year-old lens with those for an unfixed seven-year-old lens indicated that the decrease in wet weight was due mainly to a loss of water from the cortex, which resulted in virtual disappearance of the water/protein gradient and the formation of a plateau containing 58% water in over 90% of the lens. CONCLUSIONS Fixation substantially alters the amount and distribution of water in the human lens. Caution should be exercised when interpreting data on water and protein distributions as well as cell dimensions obtained with lenses which have been fixed. In addition, prolonged storage of a lens in PBS will result in substantial water uptake, which may affect measurements of their dimensions and optical properties.