• Applied Microscopy
• /
• v.47 no.2
• /
• pp.63-69
• /
• 2017

Golgi staining has been modified and developed since Camillo Golgi introduced the black reaction in 1873. This study focuses on the commonly used Golgi staining methods and presents comprehensive data regarding three Golgi staining methods along with their strong and weak points. The Golgi-Cox method uses mercuric chloride for brain tissue impregnation and is a reliable technique for analyzing the complete dendritic tree of cortical neurons. However, specimens tend to shrink during the staining steps. Recent combination of the Golgi-Cox method and immunofluorescence provides additional options for neuroscientists. Rapid Golgi staining requires osmium tetroxide for the post-fixation process. It homogenously stains whole structures of neurons and provides their detailed anatomical morphology. This staining is influenced by the age of the specimen, temperature of the laboratory, and duration of each procedure. The Golgi-Kopsch method uses formaldehyde and glutaraldehyde instead of osmium tetroxide and can be used regardless of the age of the specimen and the duration after fixation. This method is suitable for research using human brain fixed for a long time or for specimens obtained from old-aged animals. Selecting a Golgi staining protocol that is appropriate for the specimen type and research purpose is important to achieve best results.

• Journal of Korea Technical Association of The Pulp and Paper Industry
• /
• v.44 no.4
• /
• pp.16-24
• /
• 2012

To characterize the coating structure, diverse methods such as mercury intrusion, nitrogen adsorption and oil absorption methods have been developed and widely employed. These indirect techniques, however, have some limitation to explain the actual coating structure. Recently microscopic observation methods have been tried for analyzing structural characteristics of coating layers. Preparation of the undamaged cross section of a coating layer is essential for obtaining high quality image for analysis. In this study, distortion-free cross-section of the coating layer was prepared using a grinding and polishing technique. The coated paper was embedded in epoxy resin and cured. After curing the resin block it was ground with abrasive papers and then polished with diamond particle suspension and nylon cloth. Polished coating layer was sufficient enough to obtain undamaged cross sectional images with scanning electron microscope under backscattered electron image mode. In addition, the SEM images allowed distinction of the coating layer components. Also S/B latex film formed between pigment particles was visualized by osmium tetroxide staining. Pore size distribution and pore orientation were evaluated by image analysis from SEM cross-sectional images.

• Applied Microscopy
• /
• v.30 no.2
• /
• pp.113-119
• /
• 2000

Lipophilic glandular trichomes form secretory vesicles that accumulate in a distended noncellular secretory cavity . Imidazole-buffered osmium tetroxide solution was used to visualize unsaturated lipids in glands of Cannabis sativa. This method of staining revealed two kinds of secretory vesicles in the cavity of glands. Some smaller and rounded vesicles in the secretory cavity and secretory cells were positively stained with the imidazole, and they appeared electron-dense. The other type of vesicles which have bigger sizes more or less were not reacted, however, they appeared transparent. A high contrast of the cuticles which cover the gland was also strongly reacted with that processing. Those result suggest that the dark vesicles in the cavity may contribute to enlarge the subcuticular wall and cuticle when contents of these vesicles are dispersed into wall.

• Applied Microscopy
• /
• v.24 no.4
• /
• pp.61-77
• /
• 1994

The calcium-binding proteins (CaBP), parvalbumin (PV) and calbindin-D 28K (calbindin) are particularly abundant and specific in their distribution, and present in different subsets of neurons in many brain regions. Although their physiological roles in the neurons have not been elucidated, they are valuable markers of neuronal subpopulations for anatomical and developmental studies. This study is designed to characterize dorsal lateral geniculate nucleus (dLGN) neurons and axon terminals in terms of differential expression of immunoreactivity (IR) for two well-known CaBPs, PV and calbindin. The experiments were carried out on 6 adult monkeys. Monkeys were perfused under deep Nembutal anesthesia with 2% paraformaldehyde and 0.2% glutaraldehyde in 0.1M phosphate buffer. After removal, the brains were postfixed for 6-8 hr in 2% paraformaldehyde at $4^{\circ}C$ and infiltrated with 30% sucrose at $4^{\circ}C$. Thereafter, they were frozen in dry ice. Serial sections of the thalamus, at $20{\mu}m$, were made in the frontal plane with a sliding microtome. The sections were stained for PV and calbindin with indirect immunocytochemical methods. For electron microscopy, after infiltration with 30% sucrose the blocks of thalamus were serially sectioned at $50{\mu}m$ with a Vibratome in the coronal plane and stained immediately by indirect ABC methods without Triton X-100 in incubation medium. Stained sections were postfixed in 0.2% osmium tetroxide, dehydrated and flat-embedded in Spurr resin. The block was then trimmed to contain only a selected lamina or interlaminar space. The dLGN proper showed strong PV IR in fibers in all laminae and interlaminar zones. Particularly dense staining was noted in layers 1 and 2 that contain many stained fibers from optic tract. Neuronal cell body stained with PV was concentrated only in the laminae. In these laminae staining was moderate in cell bodies of all large and medium-sized neurons, and was strong in cell bodies of some small neurons together with their processes. Calbindin IR was marked in the neuronal cell body and neuropil in the S layers and interlaminar zones whereas moderate in the neuropil throughout the nucleus. Regional difference in distribution of PV and calbindin IR cell is distinct; the former is only in the laminae and the latter in both the S layer and interlaminar space. The CaBP-IR elements were confined to about $10{\mu}m$ in depth of Vibratome section. The IR product for CaBP was mainly associated with synaptic vesicle, pre- and post-synaptic membrane, and outer mitochondrial membrane and along microtubule. PV-IR was noted in various neuronal elements such as neuronal soma, dendrite, RLP, F, PSD and some myelinated or unmyelinated axons, and was not seen in the RSD and glial cells. Only a few neuronal components in dLGN was IR for calbindin and its reaction product was less dense than that of PV, and scattered throughout cytoplasm of soma of some relay neurons, and was also persent in some dendrite, myelinated axons and RLP. The RSD, F, PSD and glial elements were always non-IR for calbindin. Calbindin labelled RLP were presynaptic to unlabeled dendrite or dendritic spine and PSD. Calbindin-labeled dendrite of various sizes were always postsynaptic to unlabeled RSD, RLP or F. From this study it is suggested that dLGN cells of different functional systems and their differential projection to the visual cortex can be distinguished by differential expression of PV and calbindin.