Different Sample Preparation Methods for Glomerular Morphological Evaluation: To Coat or not to Coat?

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Priscila Fernandes dos Santos; Diogo Benchimol de Souza; Eduardo José Lopes Torres; Waldemar Silva Costa; Francisco José Barcelos Sampaio & Bianca Martins Gregorio


The renal glomerulus is coated by fenestrated endothelial cells and externally covered by specialized epithelial cells, known as podocytes. Scanning electron microscopy becomes an important and effective tool for its studies. Normally, samples destined for scanning microscopy are covered with a thin metallic layer. However, this step can be dispensed for some analyzes. We aimed to compare coated and uncoated samples for evaluation of the glomerular morphology of the Wistar rat kidney. Cortical region of the kidney of the 5- month-old male Wistar rats were used. The fragments followed the routine procedure for scanning electron microscopy processing. Half of 10 fragments were coated with palladium gold and the remaining were not coated. Auriga Compact FIB - SEM scanning electron microscope was used to observe the samples. Different increases and voltages was evaluated. For the uncoated samples, when using voltages of 2 KV (or higher) a great charging was observed, impairing the use of such voltage. Thus, these samples were always observed under voltage of 0.5 KV. On the other hand, in the coated samples, the use of 2 KV was adequate. Almost as a consequence, in the coated samples, the podocyte structures were better characterized, generating better images. Inversely, in the uncoated samples, it was possible to visualize the desired structures and to detect the morphological characteristics of these. The results showed that it is possible to use kidney samples without previous coating to evaluate the glomerular morphology at the ultrastructural level, serving as a tool in the study of pathologies.

KEY WORDS: Glomerular morphology; Renal filtration barrier; Scanning electron microscopy.

How to cite this article

DOS SANTOS, P. F.; DE SOUZA, D. B.; TORRES, E. J. L.; COSTA, W. S.; SAMPAIO, F. J. B. & GREGORIO, B. M. Different sample preparation methods for glomerular morphological evaluation: To coat or not to coat? Int. J. Morphol., 37(2):477-480, 2019.