ABSTRACT
Several new features of mitochondrial nucleoid and its surroundings in mammalian cells were described previously (Prachar, 2016). Very small details were observed using the improved transmission electron microscopy method, as described in the article. In the meantime, the method has again been improved to 2 Å resolutions in the cell section. The method described in detail in the present work is documented on the same records that were published in lower resolution in the work Prachar (2016), enabling comparison of the achieved resolution with the previous one. New records are also presented, showing extremely high resolution and thus implying the importance of the method. Potential use of this method in different fields is suggested.
Subject(s)
Cells/ultrastructure , Microscopy, Electron, Transmission/methods , Radiation Exposure/prevention & control , Animals , Cell Line, Tumor , Cells/radiation effects , Electrons , Endoplasmic Reticulum/radiation effects , Endoplasmic Reticulum/ultrastructure , Epoxy Resins , Histological Techniques , Leukemia L1210 , Mice , Mitochondria/radiation effects , Mitochondria/ultrastructure , Plastic Embedding/standards , Ribosomes/radiation effects , Ribosomes/ultrastructure , Time FactorsABSTRACT
To assess bone pathologies and bone regeneration immunohistochemistry may provide additional information compared to conventional histology. However, the effectiveness of this technique is limited due to tissue fixation, preparation and embedding. For bone tissue the standard immunohistological procedure includes formalin fixation, followed by decalcification and paraffin embedding. This may lead to a badly preserved trabecular bone structure but allows antibody application. Alternatively, methyl-methacrylate (MMA) resin may be used for embedding, thus circumventing the decalcification procedure. In this study immunohistology of typical bone markers was compared using human bone samples fixed either with alcohol or formalin and further decalcified and embedded in paraffin and decalcified or non decalcified samples embedded in Technovit 9100 New(R). On semi-thin sections immunohistochemistry with bone markers osteocalcin, osteonectin, osteopontin, collagen type I and the cellular markers CD34 and CD68 was performed. Independent of the fixative used, Technovit 9100 New embedded non-decalcified bone yielded a stronger immunostaining for all markers when compared to decalcified bone embedded either in methyl-methacrylate or paraffin. In addition there was a better preservation of the trabecular bone morphology. The immunohistochemical results demonstrate that Technovit 9100 New as a low-temperature acrylic resin embedding method can be favoured over paraffin embedding.
