Myofibrillogenesis in vitro as seen with the scanning electron microscope

The present study describes an experimental approach whereby myofibrillogenesis and the three-dimensional arrangement of myofibrils present within cultured skeletal muscle cells can be examined using the scanning electron microscope. This procedure uses cells that have been cultured on gold-coated coverslips, and treated with Triton X-100 to extract the cell membrane and the soluble cytoplasm. Subsequent electroconductive staining by treatment with thiocarbohydrazide and osmium allows the myofibrils to be visualized. The images of myofibrils in various states of development observed by this method generally accords to those previously reported by transmission electron microscopy. Cell elongation and adhesion to the substrate causes mechanical stress from different directions which meet at branchings of the cultured myotubes. Many myofibrils are observed to run in the direction of the inferred stress lines.     

Mammalian fertilization as seen with the scanning electron microscope

For several years we have been looking at mammalian gametes and their interactions with the scanning electron microscope (SEM). Examining the images produced by the SEM has given us a three-dimensional view of sperm, eggs, and egg investments. We are particularly impressed with the structural variation among gametes of different mammalian species. In this short report we examine the structure of mammalian spermatozoa, eggs, zonae pellucidae, and cumuli. Our observations and those of others have led us to believe that variation in gamete structure and function may have evolved as a mechanism for reproductive isolation of mammalian species.     

Blastoderm development in honey bee embryogenesis as seen in the scanning electron microscope

Summary

Based on observations in the scanning electron microscope, we describe the successive changes at the cell surface of fertilized honey bee eggs which, over a period of 20 h, lead to blastoderm formation. Each change starts in the differentiation center located in the anterior egg half and from there spreads as a wave towards both poles. However, one of the waves is heterogeneous: the protrusions or caps of oolemma which start enveloping the preblastoderm nuclei arise in different ways in the anterior third and in the more posterior regions of the egg cell. After the formation of these protrusions, three mitotic waves pass over the peripheral nuclei. The last of these blastemal mitoses results in a two-layered arrangement of peripheral nuclei, each surrounded by an outpocketing of the egg cell basally confluent with the central yolky part. This two-layered state after some time transforms into a single layer of columnar outpocketings which increase in height by extending their borders centripetally into the secondary periplasm (inner ‘Keimhautblastem’). The outpocketings disconnect from the central yolk mass as separate blastoderm cells only a short time before gastrulation. The events described here consume nearly 40% of the time period between oviposition and hatching; thus, and by comparison with most other insects, blastoderm formation in the honey bee is a very lengthy process.     

The surface of the ovine omasum as seen with the scanning electron microscope (Mammalia,Bovidae)

The omasal surface with a range of horny papillae and stratum corneum has been examined using scanning electron microscopy. Other features of interest described include pits or holes in the surface of these corneal cells and hair-like structures associated with the lamina.     

A morphological comparison of aortic elastin from five species as seen with the scanning electron microscope

The elastic laminae were extracted from thoracic aortas of adult animals including sheep, dogs, rabbits, cats and rats by treating them in hot alkaline solution (0.1 N NaOH at 75 degrees C) and observed with a scanning electron microscope. The elastic laminae are comprised of sheet-like internal elastic lamina, fibrous and membraneous elastin in tunica media, interlamellar fibers and hollow spaces which we presume were formerly filled with smooth muscle cells in the tunica media. These structures are the same in all five species except that the number of layers and the total thickness of the wall differs.     

Three-dimensional arrangement of sarcoplasmic reticulum in crayfish as seen by high voltage electron microscope

Summary Sections several m thick of single fibres from the crayfish muscle were examined by means of a high voltage electron microscope to find out the geometry of the sarcoplasmic reticulum (SR). A sufficient contrast of the SR was achieved by lead acetate histochemical method for calcium. Longitudinally oriented files of SR vesicles at the level of A bands and interruptions in otherwise continuous SR net are the most conspicuous features.     

Isolation and electron microscopic observations of intracytoplasmic inclusions containing Chlamydia psittaci.

Intracytoplasmic inclusions containing Chlamydia psittaci were isolated by a newly established method. Infected L-cells at 20 h after infection were suspended in 0.25 M sucrose-tris(hydroxymethyl)aminomethane buffer containing ethylene-diaminetetraacetic acid, homogenized in a Dounce tissue grinder, and filtered through a 2,000-mesh screen. Isolated inclusions were stabilized in 5% bovine serum albumin in 10 mM tris(hydroxymethyl)aminomethane buffer. Electron microscopic observations revealed the presence of surface projections on the vegetative, reticulate bodies and a direct connection between the reticulate bodies and the inclusion membrane by means of projections.