Developing formalin-based fixative agents for post mortem brain MRI at 9.4 T

Purpose To develop fixative agents for high-field MRI with suitable dielectric properties and measure MR properties in immersion-fixed brain tissue. Methods Dielectric properties of formalin-based agents were assessed (100 MHz-4.5 GHz), and four candidate fixatives with/without polyvinylpyrrolidone (PVP) and different salt concentrations were formulated. B-1 field and MR properties (T-1, R2*, R-2, R2 ', and magnetic susceptibility [QSM]) were observed in white and gray matter of pig brain samples during 0.5-35 days of immersion fixation. The kinetics were fitted using exponential functions. The immersion time required to reach maximum R2* values at different tissue depths was used to estimate the Medawar coefficient for fixative penetration. The effect of replacing the fixatives with Fluoroinert and phosphate-buffered saline as embedding media was also evaluated. Results The dielectric properties of formalin were nonlinearly modified by increasing amounts of additives. With 5% PVP and 0.04% NaCl, the dielectric properties and B-1 field reflected in vivo conditions. The highest B-1 values were found in white matter with PVP and varied significantly with tissue depth and embedding media, but not with immersion time. The MR properties depended on PVP yielding lower T-1, higher R2*, more paramagnetic QSM values, and a lower Medawar coefficient (0.9 mm/h; without PVP: 1.5). Regardless of fixative, switching to phosphate-buffered saline as embedder caused a paramagnetic shift in QSM and decreased R2* that progressed during 1 month of storage, whereas no differences were found with Fluorinert. Conclusion In vivo-like B-1 fields can be achieved in formalin fixatives using PVP and a low salt concentration, yielding lower T-1, higher R2*, and more paramagnetic QSM than without additives. The kinetics of R2* allowed estimation of fixative tissue penetration.

Automated summary

The study aimed to develop fixative agents suitable for high-field MRI and measure MR properties in immersion-fixed brain tissue. The results showed that using fixatives with polyvinylpyrrolidone and low salt concentration can achieve in vivo-like B-1 fields and higher R2* values, while replacing the embedding medium with phosphate-buffered saline leads to different effects.