Über die intrazelluläre Formation von Bakterien - DNS

1961 ◽  
Vol 16 (11) ◽  
pp. 730-739 ◽  
Author(s):  
A. Kleinschmidt. ◽  
D. Lang ◽  
C. Plescher ◽  
W. Hellmann ◽  
J. Haass ◽  
...  
Keyword(s):  
Constant Width ◽  
Inverse Function ◽  
Peripheral Zone ◽  
Section Thickness ◽  
Thin Sections ◽  
Air Interface ◽  
Optimal Section ◽  
Macromolecular System ◽  
Embedding Medium ◽  
Dna Pool

1. Electron micrographs of ultra-thin sections of Staphylococcus aureus and Micrococcus lysodeikticus in Vestopal as embedding medium disclose a multiplicity of DNA containing threads with varying interparticular distances.2. The diameter of these threads is about one tenth of the average optimal section thickness.3. This section thickness inevitably is implicated in the visualization of the internal distances between the threads as well as in some common trends in the DNA pool, a fact that has to be accounted for in the analysis of the macromolecules.4. By spreading lysozyme protoplasts of M. lysodeikticus on a water-air interface in a Langmuir trough and by transferring this surface layer to carbon supported Formvar films, two-dimensional systems can be demonstrated which as a thread of constant width comprise the total DNA content of one microorganism each.5. Such a macromolecular system shows equally shaped, coiled loops in a peripheral zone and many crossings towards the center. Branching of threads never has been observed so far. From this evidence we conlude:a) Intracellular DNA in these bacteria seems to exist in one pool as a “woolen ball” which is centered in the cytoplasm as a more or less dense object.b) This “woolen ball“ embodies the total amount of DNA most probably as one single threadlike unit.6. Partial destruction of the thread system of protoplasts will result upon changing optimal spreading conditions.7. The same kind of destruction is shown upon isolation of the DNA from protoplasts, the length of the threads being an inverse function of the number of precipitation steps showing purification.

Cell Fine Structure as Revealed in Dessicator-Dried Resinless Sections

1981 ◽  
Vol 39 ◽  
pp. 622-623
Author(s):  
R. P. Becker ◽  
J. J. Wolosewick ◽  
J. Ross-Stanton
Keyword(s):  
Fine Structure ◽  
Tannic Acid ◽  
Three Dimensional ◽  
Albino Rats ◽  
Cell Organelles ◽  
Vascular Perfusion ◽  
Thin Sections ◽  
Carbon Coated ◽  
Embedding Medium ◽  
Polyethylene Glycol 6000

Methodology has been introduced recently which allows transmission and scanning electron microscopy of cell fine structure in semi-thin sections unencumbered by an embedding medium. Images obtained from these “resinless” sections show a three-dimensional lattice of microtrabeculfee contiguous with cytoskeletal structures and membrane-bounded cell organelles. Visualization of these structures, especially of the matiiDra-nous components, can be facilitated by employing tannic acid in the fixation step and dessicator drying, as reported here.Albino rats were fixed by vascular perfusion with 2% glutaraldehyde or 1.5% depolymerized paraformaldehyde plus 2.5% glutaraldehyde in 0.1M sodium cacodylate (pH 7.4). Tissues were removed and minced in the fixative and stored overnight in fixative containing 4% tannic acid. The tissues were rinsed in buffer (0.2M cacodylate), exposed to 1% buffered osmium tetroxide, dehydrated in ethyl alcohol, and embedded in pure polyethylene glycol-6000 (PEG). Sections were cut on glass knives with a Sorvall MT-1 microtome and mounted onto poly-L-lysine, formvar-carbon coated grids while submerged in a solution of 95% ethanol containing 5% PEG.

scholarly journals CORRECTION OF POLYRIBOSOME DISTRIBUTIONS AS OBSERVED IN CELL SECTIONS BY ELECTRON MICROSCOPY

1964 ◽  
Vol 22 (3) ◽  
pp. 613-621 ◽  
Author(s):  
William Perl
Keyword(s):  
Electron Microscopy ◽  
Probability Method ◽  
Section Thickness ◽  
Thin Sections ◽  
Comparison Of The Results ◽  
Good Agreement ◽  
Rabbit Reticulocytes

Clusters of ribosomes observed by electron microscopy in thin sections of rabbit reticulocytes are of the same order of size as the section thickness of 600 A. Many of the observed clusters must therefore have been transected by the section surfaces and observed as clusters containing fewer ribosomes. A probability method of correcting for this effect is given. Comparison of the results with grid observations of ribosome distributions indicates sufficiently good agreement for application to cell section observations.

scholarly journals THE SARCOPLASMIC RETICULUM IN MUSCLE CELLS OF AMBLYSTOMA LARVAE

1956 ◽  
Vol 2 (4) ◽  
pp. 163-170 ◽  
Author(s):  
Keith R. Porter
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Sarcoplasmic Reticulum ◽  
Constant Width ◽  
Muscle Cells ◽  
Cell Types ◽  
Thin Sections ◽  
System A ◽  
Structural Relationships ◽  
Complex Membrane

Electron microscopy of thin sections of muscle fibers in myotomes of Amblystoma larvae has revealed the presence of a complex, membrane-limited system of canaliculi and vesicles which form a lace-like reticulum around and among the myofibrils. This seems to correspond to the sarcoplasmic reticulum of the earlier light microscopists and the endoplasmic reticulum of other cell types. The elements constituting the reticulum are disposed in a pattern which bears a constant relation to the bands of the adjacent myofibrils and is therefore repeated in each sarcomere. At the H band the system is transversely continuous but not so at other levels. Longitudinally continuity is interrupted at the Z bands where large vesicles belonging to adjacent sarcomere segments of the system face off on opposite sides of the band. The opposing faces of these vesicles are flat and separated by a space of more or less constant width, in which are located small, finger-shaped vesicles. In view of these and other close structural relationships with the myofibrils it seems appropriate to assign to the system a role in the conduction of the excitatory impulse.

scholarly journals Rapid embedding of tissues in Lowicryl K4M for immunoelectron microscopy.

1984 ◽  
Vol 32 (11) ◽  
pp. 1217-1223 ◽  
Author(s):  
L G Altman ◽  
B G Schneider ◽  
D S Papermaster
Keyword(s):  
Room Temperature ◽  
Dimethyl Formamide ◽  
Point Counting ◽  
Rabbit Antibody ◽  
Electron Microscope Level ◽  
Thin Sections ◽  
Kidney Microsomes ◽  
Embedding Medium ◽  
Lowicryl K4m ◽  
Accelerated Protocol

Lowicryl K4M (K4M) was recently introduced as an embedding medium for immunocytochemistry at the electron microscope level (BL Armbruster, E Carlemalm, R Chiovetti, RM Garavito, JA Hobot, E Kellenberger, W Villiger (1982):J Microsc 126:77 and E Carlemalm, M Garavito, W Villiger (1982):J Microsc 126:123). While earlier protocols of fixation and embedding required 4-6 days, the present method has reduced the processing time by accelerating both dehydration of tissues and polymerization of K4M so that tissues can be prepared for sectioning within 4 hr. The immunocytochemical labeling density was quantitated in order to determine relative antigen preservation in tissues embedded by the accelerated protocol as compared to slower K4M embedding techniques and to tissues embedded in glutaraldehyde-cross-linked bovine serum albumin (BSA). Thin sections of Bufo marinus kidney were labeled with rabbit antibody to Na+,K+ATPase alpha chain catalytic subunit isolated from B. marinus kidney microsomes (M Girardet, K Geering, JM Frantes, D Geser, BC Rossier, JP Kraehenbuhl, C Bron (1981):Biochemistry 20:6684). B. marinus retinas were labeled with rabbit anti-opsin. After fixation in paraformaldehyde(3%)-glutaraldehyde(3%), tissues were washed in buffer, dehydrated in 50, 75, and 90% dimethyl-formamide (DMF, 10 min each); K4M:DMF, 1:2 (15 min); K4M:DMF, 1:1, (20 min); K4M (25 min); K4M (30 min) at room temperature and transferred in fresh K4M to BEEM capsules for exposure to ultraviolet light (GE 15 watt, Black-lite, 10 cm, 45 min or less) at 4 degrees C. Thin sections were labeled successively with antibody, biotinylated sheep anti-rabbit F(ab')2 and avidin-ferritin. Ferritin labeling densities were determined by point counting. High labeling densities were observed with both antibodies, equaling or exceeding levels of labeling by slower protocols or embedment in BSA.

scholarly journals Improved Sectioning of Polymers Using an Oscillating Diamond Knife for Transmission Electron Microscopy

2006 ◽  
Vol 14 (5) ◽  
pp. 20-21 ◽  
Author(s):  
J.D. Harris ◽  
J.S. Vastenhout
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Wedge Angle ◽  
Cutting Speed ◽  
Sample Surface ◽  
Section Thickness ◽  
Thin Sections ◽  
Transmission Electron ◽  
Surface Section ◽  
Diamond Knife

Polymers are viscoelastic materials that can often deform during microtome sectioning. Similar to plastic embedded biological materials, many methods have been developed over the years to not only improve the image contrast of these materials but also to harden the material for improved sectioning during microtomy. Even with these improvements, a common artifact, compression, during the sectioning of this class of materials remains problematic.Compression is caused by several factors: hardness of the sample, embedding media, wedge angle of the knife, interaction between the diamond and sample surface, section thickness and cutting speed. It has been found that reducing the knife angle from 45º to 35° leads to a reduction in compression. Recent efforts to further reduce the compression of ultra-thin sections have led to the invention of an oscillating diamond knife.

scholarly journals An Effective Method of Preparing Sections of Bacillus polymyxa Sporangia and Spores for Electron Microscopy

1960 ◽  
Vol 7 (2) ◽  
pp. 373-376 ◽  
Author(s):  
Pauline E. Holbert
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Electron Micrographs ◽  
Epoxy Resins ◽  
Bacillus Polymyxa ◽  
Thin Sections ◽  
Similar Images ◽  
Embedding Medium ◽  
First Time ◽  
Diamond Knife

Bacillus polymyxa sporangia and spores were prepared for examination in the electron microscope by methods whose critical features were apparently: judicious use of vacuum, to encourage complete penetration of the embedding medium; the use of epoxy resins as embedding media; and cutting of the thin sections with a diamond knife. Electron micrographs of material prepared in this manner exhibit undeformed sporangial sections. Some of the structures revealed have been shown before, though perhaps less distinctly; other structures are revealed here for the first time. While this single study does not pretend to elucidate all the complexities of sporulation in bacteria, these and similar images should make this possible, and some mention of the preparatory techniques that lead to them seems advisable at this time.

scholarly journals Application of the avidin-biotin system for post-embedding cytochemical demonstration of a biotin-labeled IgG tracer.

1986 ◽  
Vol 34 (4) ◽  
pp. 547-549 ◽  
Author(s):  
R J Seitz ◽  
K Heininger ◽  
G Schwendemann ◽  
K V Toyka
Keyword(s):  
Skeletal Muscle ◽  
Sciatic Nerve ◽  
Blood Vessels ◽  
Tracer Studies ◽  
Thin Sections ◽  
Mouse Skeletal Muscle ◽  
Cytochemical Demonstration ◽  
Mode Of Application ◽  
Embedding Medium

The capacity of the avidin-biotin system for post-embedding cytochemical detection of a biotin-labeled proteinaceous tracer was investigated. By testing modifications of the fixatives, the epoxy embedding medium, and various etching solutions, a procedure was developed to localize specifically the biotinylated tracer on 1-micron thick and thin sections. By use of this technique, a systemically administered IgG tracer was demonstrated after 24 hr throughout the endomysium of mouse skeletal muscle. In the adjoining sciatic nerve, the tracer IgG occurred at the perineurium and within endoneurial blood vessels, although the endoneurium itself was spared because of the presence of the blood-nerve barrier. Because of the small size of the biotin ligand and the non-denaturing method of labeling proteins, our mode of application of the avidin-biotin system appears suitable for tracer studies.

scholarly journals MEASUREMENT OF THICKNESS WITHIN SECTIONS BY QUANTITATIVE ELECTRON MICROSCOPY

1969 ◽  
Vol 40 (3) ◽  
pp. 768-772 ◽  
Author(s):  
Lloyd Silverman ◽  
Berit Schreiner ◽  
David Glick
Keyword(s):  
Electron Microscopy ◽  
Phosphotungstic Acid ◽  
Exchange Resin ◽  
Interference Microscopy ◽  
Quantitative Electron Microscopy ◽  
Thin Sections ◽  
Calculated Mass ◽  
Thickness Standard ◽  
Embedding Medium ◽  
Measurement Of Thickness

To apply the method of quantitative electron microscopy to the measurement of mass in thin sections, the thickness of the section at or very near the structure to be studied must be known. Dowex anion exchange resin AG 1 x 2, stained with phosphotungstic acid (PTA) at pH 6.4, was used as a thickness standard which could be embedded and sectioned. The sectioned PTA-Dowex appeared uniformly stained and exhibited suitable electron opacity. The stoichiometry of the reaction between PTA and the Dowex resin was measured by three independent methods based on gravimetric, colorimetric, and nitrogen determinations whose results showed close agreement. From the PTA uptake, the density of the stained spheres was calculated. Mass of a defined area of PTA-Dowex was measured by quantitative electron microscopy, and from this mass and density, the volume and then the thickness were calculated. The values for thickness were compared to those obtained by interference microscopy on the embedding medium alone in the same sections.

scholarly journals THE FINE STRUCTURE OF THE NUCLEOLUS DURING MITOSIS IN THE GRASSHOPPER NEUROBLAST CELL

1965 ◽  
Vol 24 (3) ◽  
pp. 349-368 ◽  
Author(s):  
Barbara J. Stevens
Keyword(s):  
Electron Microscopy ◽  
Fine Structure ◽  
Light Microscope ◽  
Light And Electron Microscopy ◽  
Peripheral Zone ◽  
Structural Components ◽  
Thin Sections ◽  
Nucleolar Material ◽  
Central Regions ◽  
Homogeneous Background

The behavior of the nucleolus during mitosis was studied by electron microscopy in neuroblast cells of the grasshopper embryo, Chortophaga viridifasciata. Living neuroblast cells were observed in the light microscope, and their mitotic stages were identified and recorded. The cells were fixed and embedded; alternate thick and thin sections were made for light and electron microscopy. The interphase nucleolus consists of two fine structural components arranged in separate zones. Concentrations of 150 A granules form a dense peripheral zone, while the central regions are composed of a homogeneous background substance. Observations show that nucleolar dissolution in prophase occurs in two steps with a preliminary loss of the background substance followed by a dispersal of the granules. Nucleolar material reappears at anaphase as small clumps or layers at the chromosome surfaces. These later form into definite bodies, which disappear as the nucleolus grows in telophase. Evidence suggests both a collecting and a synthesizing role for the nucleolus-associated chromatin. The final, mature nucleolar form is produced by a rearrangement of the fine structural components and an increase in their mass.

Research on Reservoir Characteristics in San Zhao Peripheral Zone

2014 ◽  
Vol 986-987 ◽  
pp. 722-724
Author(s):  
Sun Fei ◽  
Qiu Han Qin ◽  
Sheng Hao Zhou ◽  
Li Peng
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Seismic Data ◽  
Peripheral Zone ◽  
Sem Analysis ◽  
Oil Reservoir ◽  
Thin Sections ◽  
Analysis Techniques ◽  
Core Logging ◽  
Scanning Electron

Using a large number of core, logging and seismic data, on the basis of sequence stratigraphy, reservoir geology, sedimentology, such as theory, the San Zhao peripheral zone were studied. FuYu reservoir is studied and Yang DaChengZi reservoir structure of reservoir. Based on coring well cores observation and rock thin section, casting thin sections and scanning electron microscopy (sem) analysis techniques, such as the SQ3–Y1 a sedimentary period main sedimentary, main lithological types, and different oil reservoir is studied.

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