Ultrastructural Features of the Oral Apparatus of Stentor

1976 ◽  
Vol 34 ◽  
pp. 142-143
Author(s):  
Elizabeth F. Howell
Keyword(s):  
Plasma Membrane ◽  
Osmium Tetroxide ◽  
Buccal Cavity ◽  
Fiber Bundles ◽  
Microtubular Root ◽  
Stentor Coeruleus ◽  
Root Fiber

The ultrastructure of the normal oral apparatus of Stentor has not been extensively studied. I report here on the ultrastructure of the buccal cavity of Stentor coeruleus.Stentor coeruleus was fixed in either a buffered mixture of osmium tetroxide and glutaraldehyde, or in buffered glutaraldehyde alone. Cells were then dehydrated and embedded in a mixture of Epon and Araldite.An extensive adoral zone of membranelles surrounds the anterior of the cell, and each membranelle consists of 2 parallel rows of cilia. These extend down into the buccal cavity. Two microtubular root fibers, or nemadesmata (Figs. 2 and 5), extend deeply into the cytoplasm from the base of each ciliary kinetosome. Mitochondria are usually closely associated with the root fiber bundles, and small vesicles are present between the nemadesmata of adjacent kinetosomes (Fig. 5). In the cytopharyngeal, non-ciliated areas of the buccal cavity, microtubular ribbons which extend into the cytoplasm are aligned perpendicular to the plasma membrane of the buccal cavity (Figs. 1 and 2).

scholarly journals The fine structure produced in cells by primary fixatives 2. Potassium dichromate

1965 ◽  
Vol s3-106 (73) ◽  
pp. 15-21
Author(s):  
JOHN R. BAKER
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Fine Structure ◽  
Golgi Apparatus ◽  
Nuclear Membrane ◽  
Osmium Tetroxide ◽  
Potassium Dichromate ◽  
Potassium Dichromate Solution ◽  
Zymogen Granules ◽  
Mouse Pancreas

The exocrine cells of the mouse pancreas were fixed in potassium dichromate solution, embedded in araldite or other suitable medium, and examined by electron microscopy. Almost every part of these cells is seriously distorted or destroyed by this fixative. The ergastoplasm is generally unrecognizable, the mitochondria and zymogen granules are seldom visible, and no sign of the plasma membrane, microvilli, or Golgi apparatus is seen. The contents of the nucleus are profoundly rearranged. It is seen to contain a large, dark, irregularly shaped, finely granular object; the evidence suggests that this consists of coagulated histone. The sole constituent of the cell that is well fixed is the inner nuclear membrane. The destructive properties of potassium dichromate are much mitigated when it is mixed in suitable proportions with osmium tetroxide or formaldehyde.

THE ORGANIZATION OF CYTOPLASMIC COMPONENTS DURING THE PHASE OF CELL WALL THICKENING IN DIFFERENTIATING CAMBIAL DERIVATIVES OF ACER RUBRUM

1965 ◽  
Vol 43 (11) ◽  
pp. 1401-1407 ◽  
Author(s):  
James Cronshaw
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Osmium Tetroxide ◽  
Acer Rubrum ◽  
Middle Layer ◽  
Wall Thickening ◽  
Cytoplasmic Microtubules ◽  
Derivatives Of ◽  
Peripheral Cytoplasm

Cambial derivatives of Acer rubrum have been examined at stages of their differentiation following fixation in 3% or 6% glutaraldehyde with a post fixation in osmium tetroxide. At early stages of development numerous free ribosomes are present in the cytoplasm, and elements of the endoplasmic reticulum tend to align themselves parallel to the cell surfaces. The plasma membrane is closely applied to the cell walls. During differentiation a complex system of cytoplasmic microtubules develops in the peripheral cytoplasm. These microtubules are oriented, mirroring the orientation of the most recently deposited microfibrils of the cell wall. The microtubules form a steep helix in the peripheral cytoplasm at the time of deposition of the middle layer of the secondary wall. During differentiation the free ribosomes disappear from the cytoplasm and numerous elements of rough endoplasmic reticulum with associated polyribosomes become more evident. In many cases the endoplasmic reticulum is associated with the cell surface. During the later stages of differentiation there are numerous inclusions between the cell wall and the plasma membrane.

scholarly journals Ultrastructural localization of alkaline phosphatase in rat eosinophil leucocytes.

1978 ◽  
Vol 26 (10) ◽  
pp. 862-864 ◽  
Author(s):  
D M Williams ◽  
J E Linder ◽  
M W Hill ◽  
R Gillett
Keyword(s):  
Alkaline Phosphatase ◽  
Plasma Membrane ◽  
Electron Microscope ◽  
Outer Surface ◽  
Human Neutrophil ◽  
Osmium Tetroxide ◽  
Diazonium Salt ◽  
Ultrastructural Localization ◽  
Microscope Examination ◽  
Membrane Location

The ultrastructural localization of alkaline phosphatase in eosinophil leucocytes, obtained from experimentally-induced peritoneal exudates in rats, has been studied using an osmiophilic technique with 2-naphthylthiolphosphoryl dichloride as substrate, fast Blue BBN as diazonium salt and postosmication with 1% aqueous osmium tetroxide. With this method identical incubation procedures could be used for both light and electron microscope examination. Eosinophils were the only cells which contained alkaline phosphatase. The enzyme was predominantly associated with the outer surface of the plasma membrane, being present in much lower concentrations in cytoplasmic cisternae. Eosinophil granules only rarely showed reaction product. The plasma membrane location of alkaline phosphatase in eosinophil leucocytes is identical to that recently demonstrated in the human neutrophil.

scholarly journals ARTIFACTUAL LOCALIZATION OF FERRITIN IN THE CILIARY EPITHELIUM IN VITRO

1965 ◽  
Vol 25 (2) ◽  
pp. 1-7 ◽  
Author(s):  
John Mcd. Tormey
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Basement Membrane ◽  
Osmium Tetroxide ◽  
Colloidal Particles ◽  
Ciliary Epithelium ◽  
Albino Rabbit ◽  
Cytoplasmic Matrix ◽  
Tracer Particles

The accumulation of ferritin by the ciliary epithelium of the adult albino rabbit has been studied by electron microscopy. The experiments have been carried out under in vitro conditions, such that any uptake observed should be the result of passive diffusion of the tracerparticles rather than the product of active metabolic processes. The cells were fixed in osmium tetroxide and embedded in Araldite. Ferritin was found localized in three areas: in rows of apparent vesicles, free in the cytoplasmic matrix, and in the basement membrane. Some of the conclusions reached are as follows. The appearance of tracer in rows of vesicles is not in itself an adequate demonstration of pinocytosis. The permeability of the plasma membrane is drastically increased by osmium tetroxide fixation, so that tracer particles are free to diffuse across the membrane and wander through the cytoplasm. These results indicate the serious danger of being misled by artifacts when colloidal particles are used as tracers.

Lomasome biogenesis and function in armillaria mellea

1978 ◽  
Vol 36 (2) ◽  
pp. 402-403
Author(s):  
B. Ch. Behboodi
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Osmium Tetroxide ◽  
Higher Plants ◽  
Cell Wall Formation ◽  
Extensive Investigation ◽  
Sodium Cacodylate ◽  
Armillaria Mellea ◽  
Synthetic Media ◽  
And Function

IntroductionBorder bodies or lomasomes are the aggregation of membranes and vesicles located between the plasma membrane and the cell wall of many fungi, algae, and higher plants. Despite extensive investigation, the biogenesis as well as function of these structures is not yet known. The purpose of this investigation was to describe the biogenesis of lomasomes in Armillaria mellea and to provide some observations on their function related to cell wall formation.Materials and MethodsVarious thalli of fungi as non-aggregated hyphae, pseudosclerotes, rhizomorphs and carpophores were grown either on orange or synthetic media as described previously. The thalli were fixed in 4% glutaraldehyde buffered with 0.1 M sodium cacodylate (pH 7.4), and 0.15 M sucrose for 4 h at 4°. They were postfixed with 1% osmium tetroxide in the same buffer for 2 h at 4° and embedded in Epon according to the Luft procedure. Cytochemical studies using thiocarbohydrazide-silver proteinate were performed according the Thiéry.

scholarly journals Subcellular localization of cellulases in auxin-treated pea.

1976 ◽  
Vol 69 (1) ◽  
pp. 97-105 ◽  
Author(s):  
A K Bal ◽  
D P Verma ◽  
H Byrne ◽  
G A Maclachlan
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Major Part ◽  
Osmium Tetroxide ◽  
Close Association ◽  
Cellulase Activity ◽  
Cytochemical Localization ◽  
Tissue Sections ◽  
Inner Surface

Two forms of cellulase, buffer soluble (BS) and buffer insoluble (BI), are induced as a result of auxin treatment of dark-grown pea epicotyls. These two cellulases have been purified to homogeneity. Antibodies raised against the purified cellulases were conjugated with ferritin and were used to localize the two cellulases. Tissue sections were fixed in cold paraformaldehyde-glutaraldehyde and incubated for 1 h in the ferritin conjugates. The sections were washed with continuous shaking for 18 h and subsequently postfixed in osmium tetroxide. Tissue incubated in unconjugated ferritin was used as a control. A major part of BI cellulase is localized at the inner surface of the cell wall in close association with microfibrils. BS cellulase is localized mainly within the distended endoplasmic reticulum. Gogli complex and plasma membrane appear to be completely devoid of any cellulase activity. These observations are consistent with cytochemical localization and biochemical data on the distribution of these two cellulases among various cell and membrane fractions.

scholarly journals Microvascular pericytes contain muscle and nonmuscle actins.

1985 ◽  
Vol 101 (1) ◽  
pp. 43-52 ◽  
Author(s):  
I M Herman ◽  
P A D'Amore
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Plasma Membrane ◽  
Fluorescence Microscopy ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Stress Fibers ◽  
Fiber Bundles

We have affinity-fractionated rabbit antiactin immunoglobulins (IgG) into classes that bind preferentially to either muscle or nonmuscle actins. The pools of muscle- and nonmuscle-specific actin antibodies were used in conjunction with fluorescence microscopy to characterize the actin in vascular pericytes, endothelial cells (EC), and smooth muscle cells (SMC) in vitro and in situ. Nonmuscle-specific antiactin IgG stained the stress fibers of cultured EC and pericytes but did not stain the stress fibers of cultured SMC, although the cortical cytoplasm associated with the plasma membrane of SMC did react with nonmuscle-specific antiactin. Whereas the muscle-specific antiactin IgG failed to stain EC stress fibers and only faintly stained their cortical cytoplasm, these antibodies reacted strongly with the fiber bundles of cultured SMC and pericytes. Similar results were obtained in situ. The muscle-specific antiactin reacted strongly with the vascular SMC of arteries and arterioles as well as with the perivascular cells (pericytes) associated with capillaries and post-capillary venules. The non-muscle-specific antiactin stained the endothelium and the pericytes but did not react with SMC. These findings indicate that pericytes in culture and in situ possess both muscle and nonmuscle isoactins and support the hypothesis that the pericyte may represent the capillary and venular correlate of the SMC.

scholarly journals ROLE OF THE GAMETE MEMBRANES IN FERTILIZATION IN SACCOGLOSSUS KOWALEVSKII (ENTEROPNEUSTA)

1963 ◽  
Vol 19 (3) ◽  
pp. 477-500 ◽  
Author(s):  
Arthur L. Colwin ◽  
Laura Hunter Colwin
Keyword(s):  
Plasma Membrane ◽  
Osmium Tetroxide ◽  
Plasma Membranes ◽  
Previous Evidence ◽  
Egg Envelope ◽  
Saccoglossus Kowalevskii ◽  
Acrosomal Membrane ◽  
Peripheral Ring ◽  
Sperm Plasma Membrane

Previous electron microscope studies of sperm-egg association in the annelid Hydroides revealed novel aspects with respect to the acrosomal region. To determine whether these aspects were unique, a comparable study was made of a species belonging to a widely separated phylum, Hemichordata. Osmium tetroxide-fixed polyspermic material of the enteropneust, Saccoglossus, was used. The acrosomal region includes the membrane-bounded acrosome, with its large acrosomal granule and shallow adnuclear invagination, and the periacrosomal material which surrounds the acrosome except at the apex; here, the acrosomal membrane lies very close to the enclosing sperm plasma membrane. After reaching the egg envelope, the spermatozoon is activated and undergoes a series of changes: the apex dehisces and around the resulting orifice the acrosomal and sperm plasma membranes form a continuous mosaic membrane. The acrosomal granule disappears. Within 7 seconds the invagination becomes the acrosomal tubule, spans the egg envelopes, and meets the egg plasma membrane. The rest of the acrosomal vesicle everts. The periacrosomal mass changes profoundly: part becomes a fibrous core (possibly equivalent to a perforatorium); part remains as a peripheral ring. The basic pattern of structure and sperm-egg association in Saccoglossus is the same as in Hydroides. Previous evidence from four other phyla as interpreted here also indicates conformity to this pattern. The major role of the acrosome is apparently to deliver the sperm plasma membrane to the egg plasma membrane.

Evaluation of blocking agents in protein A-gold immunocytochemistry

1988 ◽  
Vol 46 ◽  
pp. 386-387
Author(s):  
L. G. Komuves ◽  
E. J. King
Keyword(s):  
Plasma Membrane ◽  
Wheat Germ ◽  
Osmium Tetroxide ◽  
Protein A ◽  
L929 Cell ◽  
Blocking Agent ◽  
Gold Particles ◽  
Tissue Sections ◽  
Cacodylate Buffer ◽  
Low Levels

Various blocking agent solutions are used in protein A-gold immunocytochemistry to prevent or reduce nonspecific attachment of antibodies and/or protein A-gold particles to tissue sections. In our study of intracellular antigens present at low levels, the inadequacy of some commonly used blocking solutions became evident. We, therefore, compared several blocking solutions by exposing L929 cell cultures, at 4 °C, to a hapten-labeled lectin (dinitro-phenol-labeled wheat germ agglutinin, DNP-WGA) that binds specifically to the plasma membrane. We then used the protein A-gold method to locate the bound DNP-WGA. Since endocytosis had been prevented, gold particles not associated with the plasma membrane were regarded as nonspecific.The cells were incubated with DNP-WGA for 2 h at 4 °C, and were then fixed with 2% glutaraldehyde and 2% paraformaldehyde in 0.1 M cacodylate buffer, pH 7.2. Some samples were also postfixed with osmium tetroxide.

scholarly journals STRUCTURAL FEATURES OF MESOSOMES (CHONDRIOIDS) OF BACILLUS SUBTILIS AFTER FREEZE-ETCHING

1968 ◽  
Vol 39 (2) ◽  
pp. 251-263 ◽  
Author(s):  
N. Nanninga
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Plasma Membrane ◽  
Outer Surface ◽  
Osmium Tetroxide ◽  
Close Association ◽  
Membrane Surface ◽  
Bacterial Membranes ◽  
Inner Surface ◽  
Freeze Etching

Freeze-etched cells of Bacillus subtilis have been studied with the electron microscope. The outer surface of the plasma membrane, i.e. the side facing the cell wall, is covered with numerous granules and short strands, each measuring approximately 50 A in diameter. These strands are occasionally seen to enter the cell wall. The inner surface of the plasma membrane, i.e. the side facing the cytoplasm, appears to be sparsely dotted with small particles measuring about 50 A. The envelope of mesosomes differs from the plasma membrane. Blunt protrusions arise from its outer surface; the inner surface appears smooth. Stalked particles, as described by other investigators after negative staining with phosphotungstic acid, were not observed on any membrane surface in our material. Preparations were also made of specimens prefixed in osmium tetroxide prior to freeze-etching. Under these conditions the bacterial membranes appeared to be surprisingly well preserved. In contrast to directly frozen, unfixed cells, some osmium tetroxide-fixed preparations showed a differentiation in cytoplasm and nucleoplasm, which made it possible to observe the close association of the mesosome with the latter.

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