Environmental Scanning Electron Microscopy Gold Immunolabeling in Cell Biology

  • Francesco Rosso
  • Ferdinando Papale
  • Alfonso Barbarisi
Part of the Methods in Molecular Biology book series (MIMB, volume 931)


Immunogold labeling (IGL) technique has been utilized by many authors in combination with scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to obtain the identification/localization of receptors and antigens, both in cells and tissues. Environmental scanning electron microscopy (ESEM) represents an important tool in biomedical research, since it does not require any severe processing of the sample, lowering the risk of generating artifacts and interfere with the IGL procedure. The absence of metal coating could yield further advantages for our purpose as the labeling detection is based on the atomic number difference between nanogold spheres and the biological material. Using the gaseous secondary electron detector, compositional contrast is easily revealed by the backscattered electron component of the signal. In spite of this fact, only few published papers present a combination of ESEM and IGL. Hereby we present our method, optimized to improve the intensity and the specificity of the labeling signal, in order to obtain a semiquantitative evaluation of the labeling signal.

In particular, we used a combination of IGL and ESEM to detect the presence of a protein on the cell surface. To achieve this purpose, we chose as an experimental system 3T3 Swiss albino mouse fibroblasts and galectin-3.

Key words

Immunogold labeling Environmental scanning electron microscopy Surface protein detection Silver enhancement reaction 



This research has been funded through the Ministry of Instruction, University and Research (MIUR), number 200734RMKE: Validation of molecular markers and technology innovation in environmental scanning electron microscopy (ESEM) for the clinical and prognostic setting of clonal proliferative thyroid lesions.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Francesco Rosso
    • 1
  • Ferdinando Papale
    • 1
  • Alfonso Barbarisi
    • 1
  1. 1.IX Division of General Surgery and Applied Biotechnology, Department of Anasesthesological, Surgical and Emergency SciencesSecond University of NaplesNaplesItaly

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