ATR-FTIR Spectroscopic Analysis of Novel Fixatives and their Histological Properties On Sheep Heart

Abstract

Objective: In this study, new fixative solutions that are odorless and non-toxic were tested on the hearts of sheep to preserve the cellular structure and tissue architecture in a life-like manner. Material and Method: The changes of new fixatives before and after contact with tissue were detected for the first time by the ATR-FTIR spectroscopic method. After a preliminary inspection, hearts were fixed with six different solutions. It was periodically evaluated for weight, size, color, and hardness. Samples taken from the left vertical of each heart and stained with Hematoxylin eosin were observed histologically and alterations at the cellular level were evaluated. Results: As a result of the measurements of heart weights, reduction in Solution 1 (Standard Formaldehyde Fixative), Solution 4 and Solution 5 were detected at 5.64%, 23.70% and 14.38%, respectively. In Solution 4 and Solution 5, it was observed that the integrity of the myocardium was protected in terms of morphology, the stiffness was suited for sectioning and the coronary vessels were well preserved according to Solution 1. Further, compared to solution 1, it was determined that the endocardium and myocardium layers were well preserved in the tissues fixed in Solution 4 and Solution 5. Typical transversal streaking appearance in cardiac muscle tissue existed in all three Solution 1, Solution 4 and Solution 5. Conclusion: Solution 4 and Solution 5 were found to have superior fixative properties than Solution 1 (Formaldehyde). As a result of ATR-FTIR studies, it was determined that formaldehyde was converted into toxic formic acid in Standard Solution 1, while very few and harmless changes were detected in Solution 4 and Solution 5.