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New applications of light and electron microscopic techniques for the study of microbiological inclusions in amber

Published online by Cambridge University Press:  20 May 2016

Carmen Ascaso ,
Jacek Wierzchos ,
J. Carmelo Corral ,
Rafael López  and
Jesús Alonso
Carmen Ascaso
Affiliation:
1Centro de Ciencias Medioambientales, CSIC, Serrano 115 bis, 28006 Madrid, Spain,
Jacek Wierzchos
Affiliation:
2Servicio de Microscopía Electrónica, Universidad de Lleida, Rovira Roure 44, 25198 Lleida, Spain,
J. Carmelo Corral
Affiliation:
3Museo de Ciencias Naturales de Álava. C/Siervas de Jesús, 24. 01001 Vitoria-Gasteiz, Spain,
Rafael López
Affiliation:
3Museo de Ciencias Naturales de Álava. C/Siervas de Jesús, 24. 01001 Vitoria-Gasteiz, Spain,
Jesús Alonso
Affiliation:
3Museo de Ciencias Naturales de Álava. C/Siervas de Jesús, 24. 01001 Vitoria-Gasteiz, Spain,
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Abstract

Amber is a superb medium for the fossilization of delicate organisms. Besides light microscopy techniques for the study of insects in amber, scanning electron microscopy (SEM) in backscattered electron mode (SEM-BSE), low temperature SEM (LTSEM) and also confocal laser scanning microscopy (CLSM) were used to examine microcenosis and particulate plant remains (microdebris). We applied these techniques to such inclusions in amber Álava, northern Spain (Allaian: Early Cretaceous). Confocal microscopy provides a 3D image of partial microcenosis showing bifurcate fungal hyphae. The huge potential of SEM-BSE yields high resolution images, in which the relationship between protozoa and fungal hyphae may be observed and the characterization of further ultrastructural details in flagellates. According to the SEM-BSE images, food and pulsatile vacuoles appear better preserved than mitochondria and lipids in amber-embedded protozoa. A process of protozoan mineralization has led to the deposition of S and Fe in peripheral areas, and the Fe is also present in the core of surrounding fungal hyphae. Application of LTSEM for the study of protozoan inclusions produces images of their exteriors showing many vacuoles. Plant tissues under SEM-BSE show mummified cell walls, while the cytoplasm exhibits a bright appearance and is very rich in Fe and S. SEM in secondary electron mode (SEM-SE) also reveals a microbiota trapped in gas bubbles.

Type
Research Article
Information
Journal of Paleontology , Volume 77 , Issue 6 , November 2003 , pp. 1182 - 1192
Copyright
Copyright © The Paleontological Society 

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