Cytochemical Staining Reactions for Enzymes in Cytoplasmic Organelles

Biomembranes ◽  
1971 ◽  
pp. 33-39 ◽  
Author(s):  
Alex B. Novikoff ◽  
Phyllis M. Novikoff
Keyword(s):  
Cytochemical Staining ◽  
Cytoplasmic Organelles

Spermatogenesis in the Dragonfly with Particular Reference to the Cytoplasmic Organelles

1972 ◽  
Vol 30 ◽  
pp. 110-111
Author(s):  
Sant S. Sekhon
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Mature Sperm ◽  
Electron Microscopic ◽  
Cytoplasmic Organelles ◽  
Microscopic Studies ◽  
Insect Sperm ◽  
Large Round

Although there have been numerous studies concerning the morphogenetic changes accompanying the maturation of insect sperm, only a few deal with the sperm differentiation in the dragonflies. In two recent electron microscopic studies Kessel, has comprehensively treated the erlationship of microtubules to the nucleus and mid-piece structures during spermiogenesis in the dragonfly. The purpose of this study is to follow the sequential nuclear and cytoplasmic changes which accompany the differentiation of spermatogonium into a mature sperm during spermatogenesis in the dragonfly (Aeschna sp.).The dragonfly spermatogonia are characterized by large round nuclei. Loosely organized chromatin is usually unevenly distributed within the spermatogonial nuclei. The scant cytoplasm surrounding the nucleus contains mitochondria, the Golgi apparatus, elements of endoplasmic reticulum and numerous ribosomes (Fig. 1).

Improved immunolabelling of ultrathin cryosections using antibody conjugated with 1nm gold particles

1990 ◽  
Vol 48 (3) ◽  
pp. 928-929
Author(s):  
Morten H. Nielsen ◽  
Lone Bastholm
Keyword(s):  
Pituitary Gland ◽  
Liquid Nitrogen ◽  
Electron Density ◽  
Small Diameter ◽  
Cytochemical Staining ◽  
Gold Particles ◽  
Labelling Density ◽  
Ultrathin Cryosections ◽  
Mouse Pituitary ◽  
Electron Microscopical

During the last 5 years the diameter of the gold probes used for immuno-cytochemical staining at the electron microscopical (EM) level has been decreased. The advantage of small diameter gold probes is an overall increased labelling density. The disadvantage is a lower detectability due to the low electron density of smaller gold particles consequently an inconvenient high primary magnification needed for EM examination. Since 1 nm gold particles are barely visible by conventional EM examination the need for enlargement by silverenhancement of the gold particles has increased.In the present study of ultrathin cryosectioned material the results of immunostaining using 5 nm gold conjugated antibody and 1 nm gold conjugated antibodies are compared after silverenhancement of the 1 nm gold particles.Slices of freshly isolated mouse pituitary gland were immersion fixed for 20 min in 2 % glutaraldehyde /2 % paraformaldehyde. Blocks cryoprotected with 2.3 M sucrose were frozen in liquid nitrogen and ultra-cryosectioned on a RMC cryoultra-microtome.

The ultrastructure of oogonia and oocytes in the foetal and neonatal rat

1962 ◽  
Vol 157 (966) ◽  
pp. 99-114 ◽  
Keyword(s):  
Meiotic Prophase ◽  
Developmental Stages ◽  
Post Partum ◽  
Chromatin Condensation ◽  
Complex Form ◽  
Neonatal Rat ◽  
Mitotic Chromosomes ◽  
Thin Sections ◽  
Cytoplasmic Organelles ◽  
Spindle Fibres

Ovaries from eighty foetal and neonatal rats (aged 16·0 days post coitum to 4 days post partum ) were examined under the electron microscope. All the normal developmental stages (oogonia and oocytes in the leptotene, zygotene, pachytene, diplotene and dictyate phases of meiotic prophase) were identified. Patterns of degeneration (‘atretic divisions’, ‘ Z ’ cells and atresia at the diplotene phase), whose histological appearance and incidence have been recorded by Beaumont & Mandl (1962), were also recognized. The nuclei of oocytes at the leptotene phase contain single electron dense threads which become aligned in parallel pairs at the following phase (zygotene). A third finer fibril half-way between them appears at pachytene (tripartite ribbon). The longitudinal segments of threads, visible in ultra-thin sections, become shorter, presumably due to coiling. Nuclei at the diplotene phase contain single threads essentially similar to those seen at leptotene. As the oocyte enters the dictyate or resting phase, electron dense threads disappear from the nucleus. These observations suggest that the threads represent chromosomal ‘cores’. Nucleolus-like components persist throughout meiotic prophase, and at the diplotene phase regain the more complex form typical of oogonia. The cytoplasmic organelles become more numerous and complex as the oocyte approaches the dictyate phase. ‘Atretic divisions’ in oogonia are characterized by the persistence of long segments of nuclear membrane and the appearance of vesicles enveloped by a double membrane resembling the nuclear envelope. The dense masses of ‘chromatin’ (mitotic chromosomes) are more rounded than in normal cells at metaphase, and tend to coalesce. Spindle fibres have not been observed. Cytoplasmic organelles tend to increase in number and complexity. ‘ Z ’ cells (cells degenerating largely at the pachytene phase) show heavy ‘chromatin’ condensation around the tripartite ribbons. The major cytoplasmic changes consist in swelling of the endoplasmic reticulum, vacuolation of mitochondria and increase in incidence of multilamellar bodies. Atretic oocytes at the diplotene phase differ markedly from ‘ Z ’ cells in that ‘chromatin’ condensation around electron dense threads (single) is markedly less prominent. Cytoplasmic changes are similar to those of ‘ Z ’ cells, but also include a rise in the incidence of multivesicular and other complex bodies. All three types of degenerating cells are removed from the ovary by the phagocytic activity of neighbouring somatic cells.

scholarly journals The pattern of appearance of enzymic activity during the development of the Golgi apparatus in amoebae

1978 ◽  
Vol 34 (1) ◽  
pp. 53-63
Author(s):  
C.J. Flickinger
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Rough Endoplasmic Reticulum ◽  
Enzymic Activity ◽  
Nuclear Transplantation ◽  
Similar Distribution ◽  
Cytochemical Staining ◽  
Light Reaction ◽  
Golgi Bodies ◽  
Golgi Membranes

The appearance of enzymic activity during the development of the Golgi apparatus was studied by cytochemical staining of renucleated amoebae. In cells enucleated for 4 days, there was a great decline in size and number of Golgi bodies, or dictyosomes. Subsequent renucleation by nuclear transplantation resulted in a regeneration of Golgi bodies. Samples of amoebae were fixed and incubated for cytochemical staining at intervals of 1, 6, or 24 h after renucleation. Enzymes selected for study were guanosine diphosphatase (GDPase), esterase, and thiamine pyrophosphatase (TPPase). All three were found in the Golgi apparatus of normal amoebae but they differed in their overall intracellular distribution. GDPase was normally present at the convex pole of the Golgi apparatus, in rough endoplasmic reticulum, and in the nuclear envelope. In amoebae renucleated for 1 h, light reaction product for GDPase was present throughout the small stacks of cisternae that represented the forming Golgi apparatus. By 6 h following the operation GDPase reaction product was concentrated at the convex pole of the Golgi apparatus. Esterase, which was distributed throughout the stacks of normal Golgi cisternae, displayed a similar distribution in the forming Golgi bodies as soon as they were visible. TPPase was normally present in the Golgi apparatus but was not found in the endoplasmic reticulum. In contrast to the other enzymes, TPPase reaction product was absent from the forming Golgi apparatus 1 and 6 h after renucleation, and did not appear in the Golgi apparatus until 24 h after operation. Thus, enzymes held in common between the rough endoplasmic reticulum and the Golgi apparatus were present in the forming Golgi apparatus as soon as it was detectable, but an enzyme cytochemically localized to the Golgi apparatus only appeared later in development of the organelle. It is suggested that Golgi membranes might be derived from the endoplasmic reticulum and thus immediately contain endoplasmic reticulum enzymes, while Golgi-specific enzymes are added later in development.

scholarly journals Hematological and cytochemical characteristics of peripheral blood cells in the argus snakehead (Ophiocephalus argus Cantor)

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11234
Author(s):  
Xue Wang ◽  
Zhengjie Wu ◽  
Shengmei Wu ◽  
Xianxian Chen ◽  
Misbah Hanif ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Blood Cells ◽  
Large Scale ◽  
Periodic Acid ◽  
Economic Value ◽  
Chloroacetic Acid ◽  
Peripheral Blood Cells ◽  
Cytochemical Staining ◽  
Periodic Acid Schiff ◽  
Natural Defense

Background The argus snakehead (Ophiocephalus argus Cantor) is a highly nutritious, freshwater, cultured bony fish with a high economic value. The health of the fish is closely related to its blood cells, which are critical for oxygen transport, natural defense, and immunity. We investigated the morphometry, microstructure, and cytochemical characteristics of the peripheral blood cells of O. argus. Our results may provide the basic reference values needed to monitor the health of this fish for large-scale cultivation. Methods The number of blood cells in O. argus were counted on a hemocytometer and their size was measured using a micrometer under light microscope. The morphology and classification of the blood cells were studied using Wright’s staining and the cytochemical characteristics were studied using seven chemical stains including peroxidase (POX), Sudan black B (SBB), periodic acid-Schiff (PAS), acid phosphatase (ACP), alkaline phosphatase (ALP), chloroacetic acid AS-D naphthol esterase (AS-D), and α-naphthol acetate esterase (α-NAE). Results The peripheral blood cells in O. argus can be classified as erythrocytes, leukocytes, and thrombocytes; of which, females had 2.9597 million/mm3, 88,400/mm3, and 43,600/mm3, respectively, and males had 3.0105 million/mm3, 105,500/mm3, and 34,000/mm3, respectively. Leukocytes consisted of neutrophils, monocytes, large lymphocytes, and small lymphocytes. Eosinophils and basophils were not found. Monocytes were the most numerous leukocytes identified, followed by neutrophils and small lymphocytes, while large lymphocytes were the least frequently identified. Cytochemical staining showed that erythrocytes were only positive for PAS staining. Neutrophils were strongly positive for POX, SBB, and ACP, and positive for all the other cytochemical stains. Monocytes were positive for PAS and α-NAE and were weakly positive for ACP and AS-D staining. Large lymphocytes were positive for PAS and were weakly positive for ALP, AS-D, and α-NAE staining. Small lymphocytes were positive for PAS and weakly positive for AS-D and α-NAE staining. Thrombocytes were positive for PAS and were weakly positive for ACP and AS-D, but negative for the remaining cytochemical stains. The morphology of peripheral blood cells in O. argus was generally similar to that of other fish species, while the cytochemical staining patterns showed clear species specificity.

scholarly journals Intracellular localization of Rickettsia tsutsugamushi in polymorphonuclear leukocytes.

1979 ◽  
Vol 150 (3) ◽  
pp. 703-708 ◽  
Author(s):  
Y Rikihisa ◽  
S Ito
Keyword(s):  
Guinea Pig ◽  
Cell Cultures ◽  
Polymorphonuclear Leukocyte ◽  
Polymorphonuclear Leukocytes ◽  
Intracellular Localization ◽  
The Other ◽  
Cytoplasmic Organelles

Rickettsia tsutsugamushi (Gilliam strain) was serially propagated in BHK-21 cell cultures and incubated with guinea pig peritoneal polymorphonuclear leukocytes to study the ultrastructural features of rickettsial uptake and entry into the leukocytes. Significant numbers of rickettsiae were phagocytized selectively by these leukocytes within 30 min. About one-half of these rickettsiae remained sequestered in phagosomes but the other one-half were free from the phagosome and localized directly in the polymorphonuclear leukocyte cytoplasm. Various stages of rickettsial release from the phagosomes were observed. Once free within the polymorphonuclear leukocyte cytoplasm, the rickettsiae were preferentially localized in the glycogen-packed areas which are devoid of lysosomes and other cytoplasmic organelles. This study indicates that rickettsiae phagocytized by polymorphonuclear leukocytes can escape from the phagosome into the cytoplasm.

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