scholarly journals Microscopical and elemental FESEM and Phenom ProX-SEM-EDS analysis of osteocyte- and blood vessel-like microstructures obtained from fossil vertebrates of the Eocene Messel Pit, Germany

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1618 ◽  
Author(s):  
Edwin Cadena

The Eocene (∾48 Ma) Messel Pit in Germany is a UNESCO World Heritage Site because of its exceptionally preserved fossils, including vertebrates, invertebrates, and plants. Messel fossil vertebrates are typically characterized by their articulated state, and in some cases the skin, hair, feathers, scales and stomach contents are also preserved. Despite the exceptional macroscopic preservation of Messel fossil vertebrates, the microstructural aspect of these fossils has been poorly explored. In particular, soft tissue structures such as hair or feathers have not been chemically analyzed, nor have bone microstructures. I report here the preservation and recovery of osteocyte-like and blood vessel-like microstructures from the bone of Messel Pit specimens, including the turtlesAllaeochelys crassesculptaandNeochelys franzeni, the crocodileDiplocynodon darwini, and the pangolinEomanis krebsi. I used a Field Emission Scanning Electron Microscope (FESEM) and a Phenom ProX desktop scanning electron microscope (LOT-QuantumDesign) equipped with a thermionic CeB6 source and a high sensitivity multi-mode backscatter electron (BSE) for microscopical and elemental characterization of these bone microstructures. Osteocyte-like and blood vessel-like microstructures are constituted by a thin layer (∾50 nm thickness), external and internal mottled texture with slightly marked striations. Circular to linear marks are common on the external surface of the osteocyte-like microstructures and are interpreted as microbial troughs. Iron (Fe) is the most abundant element found in the osteocyte-like and blood vessel-like microstructures, but not in the bone matrix or collagen fibril-like microstructures. The occurrence of well-preserved soft-tissue elements (at least their physical form) establishes a promising background for future studies on preservation of biomolecules (proteins or DNA) in Messel Pit fossils.

Author(s):  
F. Ouyang ◽  
D. A. Ray ◽  
O. L. Krivanek

Electron backscattering Kikuchi diffraction patterns (BKDP) reveal useful information about the structure and orientation of crystals under study. With the well focused electron beam in a scanning electron microscope (SEM), one can use BKDP as a microanalysis tool. BKDPs have been recorded in SEMs using a phosphor screen coupled to an intensified TV camera through a lens system, and by photographic negatives. With the development of fiber-optically coupled slow scan CCD (SSC) cameras for electron beam imaging, one can take advantage of their high sensitivity and wide dynamic range for observing BKDP in SEM.We have used the Gatan 690 SSC camera to observe backscattering patterns in a JEOL JSM-840A SEM. The CCD sensor has an active area of 13.25 mm × 8.83 mm and 576 × 384 pixels. The camera head, which consists of a single crystal YAG scintillator fiber optically coupled to the CCD chip, is located inside the SEM specimen chamber. The whole camera head is cooled to about -30°C by a Peltier cooler, which permits long integration times (up to 100 seconds).


Author(s):  
H. Todokoro ◽  
S. Fukuhara ◽  
Y. Sakitani

A low acceleration scanning electron microscope ( LASEM ) offers several advantages: no charging effects low radiation damage high yield for secondary electron emission high sensitivity for surface topography The application of a low acceleration microscope, however, has been limited to special purposes because of its poor resolution. A high resolution LASEM has been developed and is shown in Fig.1. The microscope uses a new electron gun with a field emission cathode. The gun 1,000 times brighter than a conventional thermionic cathode. The relation between brightness and resolution of a SEM for 1 kV acceleration voltage is shown in Fig.2. The three regions in the figure correspond to tungsten thermionic, LaB6 thermionic and field emission guns. Resolution is approximately 1 μm in the case of the tungsten thermionic gun at 1 kV, while resolution is 200 A in the new microscope equipped with a field emission gun.


Author(s):  
R. E. Ferrell ◽  
G. G. Paulson

The pore spaces in sandstones are the result of the original depositional fabric and the degree of post-depositional alteration that the rock has experienced. The largest pore volumes are present in coarse-grained, well-sorted materials with high sphericity. The chief mechanisms which alter the shape and size of the pores are precipitation of cementing agents and the dissolution of soluble components. Each process may operate alone or in combination with the other, or there may be several generations of cementation and solution.The scanning electron microscope has ‘been used in this study to reveal the morphology of the pore spaces in a variety of moderate porosity, orthoquartzites.


Author(s):  
C. T. Nightingale ◽  
S. E. Summers ◽  
T. P. Turnbull

The ease of operation of the scanning electron microscope has insured its wide application in medicine and industry. The micrographs are pictorial representations of surface topography obtained directly from the specimen. The need to replicate is eliminated. The great depth of field and the high resolving power provide far more information than light microscopy.


Author(s):  
K. Shibatomi ◽  
T. Yamanoto ◽  
H. Koike

In the observation of a thick specimen by means of a transmission electron microscope, the intensity of electrons passing through the objective lens aperture is greatly reduced. So that the image is almost invisible. In addition to this fact, it have been reported that a chromatic aberration causes the deterioration of the image contrast rather than that of the resolution. The scanning electron microscope is, however, capable of electrically amplifying the signal of the decreasing intensity, and also free from a chromatic aberration so that the deterioration of the image contrast due to the aberration can be prevented. The electrical improvement of the image quality can be carried out by using the fascionating features of the SEM, that is, the amplification of a weak in-put signal forming the image and the descriminating action of the heigh level signal of the background. This paper reports some of the experimental results about the thickness dependence of the observability and quality of the image in the case of the transmission SEM.


Author(s):  
S. Takashima ◽  
H. Hashimoto ◽  
S. Kimoto

The resolution of a conventional transmission electron microscope (TEM) deteriorates as the specimen thickness increases, because chromatic aberration of the objective lens is caused by the energy loss of electrons). In the case of a scanning electron microscope (SEM), chromatic aberration does not exist as the restrictive factor for the resolution of the transmitted electron image, for the SEM has no imageforming lens. It is not sure, however, that the equal resolution to the probe diameter can be obtained in the case of a thick specimen. To study the relation between the specimen thickness and the resolution of the trans-mitted electron image obtained by the SEM, the following experiment was carried out.


Author(s):  
R. F. Schneidmiller ◽  
W. F. Thrower ◽  
C. Ang

Solid state materials in the form of thin films have found increasing structural and electronic applications. Among the multitude of thin film deposition techniques, the radio frequency induced plasma sputtering has gained considerable utilization in recent years through advances in equipment design and process improvement, as well as the discovery of the versatility of the process to control film properties. In our laboratory we have used the scanning electron microscope extensively in the direct and indirect characterization of sputtered films for correlation with their physical and electrical properties.Scanning electron microscopy is a powerful tool for the examination of surfaces of solids and for the failure analysis of structural components and microelectronic devices.


Author(s):  
S. Saito ◽  
H. Todokoro ◽  
S. Nomura ◽  
T. Komoda

Field emission scanning electron microscope (FESEM) features extremely high resolution images, and offers many valuable information. But, for a specimen which gives low contrast images, lateral stripes appear in images. These stripes are resulted from signal fluctuations caused by probe current noises. In order to obtain good images without stripes, the fluctuations should be less than 1%, especially for low contrast images. For this purpose, the authors realized a noise compensator, and applied this to the FESEM.Fig. 1 shows an outline of FESEM equipped with a noise compensator. Two apertures are provided gust under the field emission gun.


Author(s):  
Emil Bernstein

An interesting method for examining structures in g. pig skin has been developed. By modifying an existing technique for splitting skin into its two main components—epidermis and dermis—we can in effect create new surfaces which can be examined with the scanning electron microscope (SEM). Although this method is not offered as a complete substitute for sectioning, it provides the investigator with a means for examining certain structures such as hair follicles and glands intact. The great depth of field of the SEM complements the technique so that a very “realistic” picture of the organ is obtained.


Author(s):  
C.V.L. Powell

The overall fine structure of the eye in Placopecten is similar to that of other scallops. The optic tentacle consists of an outer columnar epithelium which is modified into a pigmented iris and a cornea (Fig. 1). This capsule encloses the cellular lens, retina, reflecting argentea and the pigmented tapetum. The retina is divided into two parts (Fig. 2). The distal retina functions in the detection of movement and the proximal retina monitors environmental light intensity. The purpose of the present study is to describe the ultrastructure of the retina as a preliminary observation on eye development. This is also the first known presentation of scanning electron microscope studies of the eye of the scallop.


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