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Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse
Published on: October 26, 2017
Influence of different fixation protocols on the preservation and dimensions of cardiac tissue
Mateusz K Hołda1, Wiesława Klimek-Piotrowska1, Mateusz Koziej1
1Department of Anatomy, Jagiellonian University Medical College, Cracow, Poland.
Insights
Formaldehyde fixation alters cardiac tissue dimensions, with 10% formaldehyde solution showing minimal changes. For accurate heart morphometric studies, measurements should be taken after one week of preservation.
Area of Science:
- Anatomy
- Histology
- Biomedical Engineering
Background:
- Cardiac morphometric studies are crucial for invasive cardiac procedures.
- Formaldehyde fixation is commonly used, but its effect on tissue dimensions is poorly understood.
- Understanding fixation-induced dimensional changes is vital for accurate anatomical measurements.
Purpose of the Study:
- To investigate the impact of different formaldehyde fixatives and concentrations on cardiac tissue dimensions.
- To determine the optimal fixation time for reliable morphometric measurements.
- To identify the best fixative for preserving cardiac tissue dimensions.
Main Methods:
- 40 pig hearts were used, with seven sites marked per heart.
- Samples were divided into four groups using 2%, 4% (formalin), 10% formaldehyde, and alcoholic formalin.
- Measurements were taken before fixation and at 24, 72, and 168 hours post-fixation.
Main Results:
- Different fixatives significantly affected various cardiac parameters.
- Most dimensions stabilized within 24 hours, with minimal changes thereafter.
- 10% formaldehyde showed the smallest dimensional changes, particularly after 168 hours.
Conclusions:
- 10% formaldehyde phosphate-buffered solution is the preferred fixative for cardiac morphometry due to minimal dimensional alterations.
- Measurements should ideally be performed after one week of tissue preservation for maximum stability.
- Accurate anatomical measurements require careful consideration of fixation methods and timing.
Abstract:
Recent extensive progress in invasive cardiac procedures has triggered a wave of dozens of heart morphometric anatomical studies that are carried out largely using autopsied samples fixed in formaldehyde solution prior to observations and measurements. In reality, very little is known about changes in heart tissue dimensions during fixation. The aim of this study was therefore to investigate how fixation affects the dimensions of cardiac tissue, and if different types and concentrations of reagents affect this phenomenon. A total of 40 pig heart samples were investigated, and seven different measuring sites were permanently marked in every heart prior to fixation. Four study groups (n = 10 each) were assembled that differed only in concentration and the type of fixative: (i) 2% formaldehyde solution; (ii) 4% formaldehyde solution (formalin); (iii) 10% formaldehyde solution; (iv) alcoholic formalin. The samples were measured before and after fixation at the following time points: 24 h, 72 h and 168 h. It was found that different fixatives significantly affected different parameters. Almost all of the heart dimensions that were measured stabilized after 24 h; later changes were statistically insignificant in the point-to-point comparison. Change in the length of the interatrial septum surface was not altered significantly in any of the fixatives after 24 h of preservation. It was found that 10% formaldehyde increased the thickness of muscular tissue only after 24 h; this thickening was reduced after 72 h and was insignificant at 168 h. Other heart parameters in this group do not present significant changes over the entire fixation time duration. In conclusion, the 10% formaldehyde phosphate-buffered solution appeared to be the best fixative among the fixatives that were studied for cardiac morphometric purposes; this solution caused the smallest changes in tissue dimensions. Measurements should be obtained at least after 1 week of preservation when most parameters exhibit the smallest changes compared with the non-preserved samples.
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