Study of the porosity of calcified chicken eggshell using atomic force microscopy and image processing

I Arzate-Vázquez; J V Méndez-Méndez; E A Flores-Johnson; J Nicolás-Bermúdez; J J Chanona-Pérez; E Santiago-Cortés

doi:10.1016/j.micron.2018.12.008

Study of the porosity of calcified chicken eggshell using atomic force microscopy and image processing

Micron. 2019 Mar:118:50-57. doi: 10.1016/j.micron.2018.12.008. Epub 2018 Dec 21.

Authors

I Arzate-Vázquez¹, J V Méndez-Méndez², E A Flores-Johnson³, J Nicolás-Bermúdez⁴, J J Chanona-Pérez⁴, E Santiago-Cortés⁴

Affiliations

¹ Centro de Nanociencias y Micro y Nanotecnologías, Instituto Politécnico Nacional, Luis Enrique Erro s/n, Zacatenco, 07738, Gustavo A. Madero, Ciudad de México, Mexico.
² Centro de Nanociencias y Micro y Nanotecnologías, Instituto Politécnico Nacional, Luis Enrique Erro s/n, Zacatenco, 07738, Gustavo A. Madero, Ciudad de México, Mexico. Electronic address: jmendezm@ipn.mx.
³ CONACYT - Unidad de Materiales, Centro de Investigación Científica de Yucatán, Calle 43, No. 130, Chuburná de Hidalgo, Mérida, 97205, Yucatán, Mexico.
⁴ Departamento de Ingeniería Bioquímica, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Av. Wilfrido Massieu Esq., Cda. Miguel Stampa s/n, 07738, Gustavo A. Madero, Ciudad de México, Mexico.

PMID: 30590254
DOI: 10.1016/j.micron.2018.12.008

Abstract

In this work, the porosity of the layers of calcified chicken eggshell (vertical crystal layer VCL, palisade layer PL and mammillary layer ML) was evaluated using atomic force microscopy (AFM) and image processing (IP). AFM topographic images were obtained from different locations for each layer and along the cross-section of calcified eggshell. Roughness parameters, surface area values, pore size and shape, surface porosity, area occupied by pores and pore density were obtained from AFM and IP. It was observed that the thickest layer (PL) exhibited the highest degree of porosity (surface porosity = 2.75 ± 1.68%, pore density = 162 ± 60 pores/μm²) when compared to the other two layers. In general, the pores located in all layers ("bubble pores") had circular shape and similar sizes. Measurements revealed a porosity gradient along the cross-section which varied with position, i.e., increasing surface porosity from the VCL towards the region of the PL closer to the ML, and decreasing surface porosity from this location towards the ML innermost surface. This suggests that the calcified eggshell has a sandwich-like structure where porosity may influence gas exchange and mechanical properties. The combination of AFM with IP presented here provides a simple and precise method to study porosity in calcified chicken eggshell, and this methodology could be used to examine other types of porous biological materials.

Keywords: Atomic force microscopy; Calcified chicken eggshell; Image processing; Porosity.

Publication types

Research Support, Non-U.S. Gov't