Objectives: To investigate the penetration depth of formaldehyde-based fixatives into cardiac muscle samples over the course of fixation.
Background: Fixation is the essential step in anatomical studies. However, very little is known about penetration of most common fixatives into cardiac tissue.
Methods: A total of 40 heart samples were investigated. 4 study groups (n=10 in each case) were formed in such manner they differed only in concentration and type of fixative (1) - 2% formaldehyde phosphate-buffered solution (FPBS); (2) - 4% FPBS (formalin); (3) - 10% FPBS; (4) - alcoholic formalin. Samples were measured before fixation and in the following time points: 24 hours, 72 hours, 168 hours.
Results: The penetration depth differed significantly among studied fixatives (p<0.0001). 100% penetration occurred in all samples after 72 hours in alcoholic formalin solution and after 168 hours in 10% FPBS. After alcoholic formalin fixation, the tissue is more brittle and sub-epicardial blisters were observed in some cases.
Conclusions: Alcoholic formalin solution is the fastest fixative among the studied ones, however it has several adverse effects on tissue structure. It was found that 10% FPBS is the best and a relatively fast fixative for cardiac morphometric studies.
Keywords: alcoholic formalin; fixation; formaldehyde; formalin; heart anatomy; morphometry.