Introduction: Buffer Variants in SEM
In the domain of scanning electron microscopy (SEM), the choice of buffer profoundly influences tissue fixation and morphology. This study focuses on comparing the effects of two common buffers, Na-cacodylate and PIPES, on SEM outcomes in tissues fixed with 3% glutaraldehyde.
Experimental Design: Buffer Influence on Frog and Chicken Embryos
Frog and chicken embryos, chosen for their delicate nature and lipid-rich content, underwent fixation in 0.03 M and 0.1 M PIPES (pH 7.3) or 0.1 M Na-cacodylate (pH 7.3). The uniformity of embryo size ensured consistent fixation quality across samples.
Analytical Focus: Lipid Preservation and Buffer Efficacy
Post-fixation, lipid preservation was assessed through extraction and analysis. Remarkably, PIPES buffer demonstrated significantly reduced lipid losses compared to Na-cacodylate. This finding was substantiated by observed morphological disparities in both SEM and transmission electron microscopy (TEM).
Conclusion: Enhancing SEM Precision with PIPES Buffer
The study highlights the pivotal role of PIPES buffer in optimizing SEM results, particularly in preserving delicate tissues’ lipid content. By selecting appropriate buffers, researchers can enhance the accuracy and reliability of SEM, opening avenues for nuanced cellular studies and advancing the field of microscopy.