Petri Dish-ELISA, a Simple and Economic Technique for Detecting Plant Viruses

2001 ◽  
Vol 36 (1-2) ◽  
pp. 1-8 ◽  
Author(s):  
M. E. Abdalla ◽  
S. E. Albrechtsen
Keyword(s):  
Petri Dish ◽  
Plant Viruses

The Fine Structure of Rat Granulosa Cell Cultures Correlated with Progestin Secretion

1973 ◽  
Vol 31 ◽  
pp. 552-553
Author(s):  
T. M. Crisp ◽  
F.R. Denys
Keyword(s):  
Fine Structure ◽  
Granulosa Cell ◽  
Cell Cultures ◽  
Single Injection ◽  
Surrounding Medium ◽  
Petri Dish ◽  
Female Rats ◽  
Vaginal Smear ◽  
Immature Female ◽  
Serum Gonadotropin

The purpose of this paper is to present observations on the fine structure of rat granulosa cell cultures grown in the presence of an adenohypophyseal explant and to correlate the morphology of these cells with progestin secretion. Twenty-six day old immature female rats were given a single injection of 5 IU pregnant mares serum gonadotropin (PMS) in order to obtain ovaries with large vesicular follicles. At 66 hrs. post-PMS administration (estrus indicated by vaginal smear cytology), the ovaries were removed and placed in a petri dish containing medium 199 and 100 U penicillin/streptomycin (P/S)/ml. Under a 20X magnification dissecting microscope, some 5-8 vesicular follicles/ovary were punctured and the granulosa cells were expressed into the surrounding medium. The cells were transferred to centrifuge tubes and spun down at 1000 rpm for 5 mins.

Ectodesmata as Revealed By Freeze-Etch Preparations of Plant Epidermal Cell Walls

1973 ◽  
Vol 31 ◽  
pp. 468-469
Author(s):  
N.C. Lyon ◽  
W. C. Mueller
Keyword(s):  
Electron Microscopy ◽  
Acetic Acid ◽  
Nitric Acid ◽  
Oxalic Acid ◽  
Light Microscopy ◽  
Epidermal Cell ◽  
Cell Walls ◽  
Plant Viruses ◽  
Plant Leaves ◽  
Thin Sections

Schumacher and Halbsguth first demonstrated ectodesmata as pores or channels in the epidermal cell walls in haustoria of Cuscuta odorata L. by light microscopy in tissues fixed in a sublimate fixative (30% ethyl alcohol, 30 ml:glacial acetic acid, 10 ml: 65% nitric acid, 1 ml: 40% formaldehyde, 5 ml: oxalic acid, 2 g: mecuric chloride to saturation 2-3 g). Other workers have published electron micrographs of structures transversing the outer epidermal cell in thin sections of plant leaves that have been interpreted as ectodesmata. Such structures are evident following treatment with Hg++ or Ag+ salts and are only rarely observed by electron microscopy. If ectodesmata exist without such treatment, and are not artefacts, they would afford natural pathways of entry for applied foliar solutions and plant viruses.

Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

1984 ◽  
Vol 42 ◽  
pp. 226-227 ◽  
Author(s):  
K. Pegg-Feige ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Immunoelectron Microscopy ◽  
Detection Method ◽  
Solid Phase ◽  
Protein A ◽  
Plant Viruses ◽  
Rapid Diagnosis ◽  
Virus Diagnosis ◽  
Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

Rotary Beveling for In Situ Thin-Section Microscopy of Cell Cultures

1981 ◽  
Vol 39 ◽  
pp. 550-551
Author(s):  
Dean A. Handley ◽  
Jack T. Alexander ◽  
Shu Chien
Keyword(s):  
Cell Growth ◽  
Cell Cultures ◽  
Molecular Interactions ◽  
Convenient Method ◽  
Petri Dish ◽  
Simple Method ◽  
Thin Section Microscopy ◽  
Thin Sectioning ◽  
Organic Fluids

In situ preparation of cell cultures for ultrastructural investigations is a convenient method by which fixation, dehydration and embedment are carried out in the culture petri dish. The in situ method offers the advantage of preserving the native orientation of cell-cell interactions, junctional regions and overlapping configurations. In order to section after embedment, the petri dish is usually separated from the polymerized resin by either differential cryo-contraction or solvation in organic fluids. The remaining resin block must be re-embedded before sectioning. Although removal of the petri dish may not disrupt the native cellular geometry, it does sacrifice what is now recognized as an important characteristic of cell growth: cell-substratum molecular interactions. To preserve the topographic cell-substratum relationship, we developed a simple method of tapered rotary beveling to reduce the petri dish thickness to a dimension suitable for direct thin sectioning.

Unlabeled Antibody Immunocytochemistry Applied to Murine Mammary Tumor Cells

1975 ◽  
Vol 33 ◽  
pp. 390-391
Author(s):  
Wm. J. Arnold ◽  
J. Russo ◽  
H. D. Soule ◽  
M. A. Rich
Keyword(s):  
Horseradish Peroxidase ◽  
Mammary Tumor ◽  
Covalent Binding ◽  
Petri Dish ◽  
Gamma Chain ◽  
Mammary Tumor Virus ◽  
Mammary Tumor Cells ◽  
Murine Mammary Tumor ◽  
Tumor Virus ◽  
Unlabeled Antibody

Our studies of mammary tumor virus have included the application of the unlabeled antibody enzyme method of Sternberger to mammary tumor derived mouse cells in culture and observation with an electron microscope. The method avoids the extravagance of covalent binding of indicator molecules (horseradish peroxidase) with precious antibody locator molecules by relying instead upon specific antibody-antigen linkages. Our reagents included: Primary Antibody, rabbit anti-murine mammary tumor virus (MuMTV) which was antiserum 113 AV-2; Secondary Antibody, goat anti-rabbit IgG gamma chain (Cappel Laboratories); andthe Indicator, rabbit anti-horseradish peroxidase - horseradish peroxidase complex (PAP) (Cappel Labs.). Dilutions and washes were made in 0.05 M Tris 0.15 M saline buffered to pH 7.4. Cell monolayers, after light fixation in glutaraldehyde, were incubated in place by a protocol adapted from Sternberger and Graham and Karnovsky, then embedded by our usual method for monolayers. Reagents were confined to specific areas by neoprene 0-rings (Parker Seal Co.) reducing the amount of reagent needed to 50 microliters, 1/6th of that required to wet a 35 mm petri dish.

A freeze-fracture-etched study of the physarum polycephalum plasma membrane during early formation of the macroplasmodial stage

1993 ◽  
Vol 51 ◽  
pp. 346-347
Author(s):  
Randolph W. Taylor ◽  
Henrie Treadwell
Keyword(s):  
Plasma Membrane ◽  
Life Cycle ◽  
Growth Phase ◽  
Filter Paper ◽  
Physarum Polycephalum ◽  
Freeze Fracture ◽  
Petri Dish ◽  
Wire Screen ◽  
Wide Mouth ◽  
Sterile Filter

The plasma membrane of the Slime Mold, Physarum polycephalum, process unique morphological distinctions at different stages of the life cycle. Investigations of the plasma membrane of P. polycephalum, particularly, the arrangements of the intramembranous particles has provided useful information concerning possible changes occurring in higher organisms. In this report Freeze-fracture-etched techniques were used to investigate 3 hours post-fusion of the macroplasmodia stage of the P. polycephalum plasma membrane.Microplasmodia of Physarum polycephalum (M3C), axenically maintained, were collected in mid-expotential growth phase by centrifugation. Aliquots of microplasmodia were spread in 3 cm circles with a wide mouth pipette onto sterile filter paper which was supported on a wire screen contained in a petri dish. The cells were starved for 2 hrs at 24°C. After starvation, the cells were feed semidefined medium supplemented with hemin and incubated at 24°C. Three hours after incubation, samples were collected randomly from the petri plates, placed in plancettes and frozen with a propane-nitrogen jet freezer.

A Sensitive On-Grid Immunoelectron Microscopic Procedure for Diagnosis of Rotavirus Infection

1980 ◽  
Vol 38 ◽  
pp. 506-507
Author(s):  
M. F. Miller ◽  
A. R. Rubenstein
Keyword(s):  
Electron Microscopy ◽  
Specific Antibody ◽  
Immune Complexes ◽  
Acute Gastroenteritis ◽  
Plant Viruses ◽  
Aggregation Process ◽  
Microscopic Technique ◽  
Fecal Material ◽  
Present Communication ◽  
Number Of Particles

Studies of rotavirus particles in humans, monkeys and various non-primates with acute gastroenteritis have involved detection of virus in fecal material by electron microscopy. The EM techniques most commonly employed have been the conventional negative staining (Fig. 1) and immune aggregation (Fig. 2) procedures. Both methods are somewhat insensitive and can most reliably be applied to samples containing large quantities of virus either naturaLly or as a result of concentration by ultracentrifugation. The formation of immune complexes by specific antibody in the immune aggregation procedures confirms the rotavirus diagnosis, but the number of particles per given microscope field is effectively reduced by the aggregation process. In the present communication, we describe use of an on-grid immunoelectron microscopic technique in which rotavirus particles are mounted onto microscope grids that were pre-coated with specific antibody. The technique is a modification of a method originalLy introduced by Derrick (1) for studies of plant viruses.

The influence of different types of milking units on milk quality and safety

2019 ◽  
pp. 118-125
Author(s):  
A. Vovkohon ◽  
V. Nadtochiy ◽  
G. Kalinina ◽  
O. Hrebelnyk ◽  
N. Fedoruk ◽  
...  
Keyword(s):  
Mass Fraction ◽  
Dry Matter ◽  
Freezing Point ◽  
Titratable Acidity ◽  
Petri Dish ◽  
Milk Quality ◽  
Quality And Safety ◽  
Protein Mass ◽  
Seeding Method ◽  
Conductivity Increase

The article highlights comparative research results of milk quality indices obtained from the milking in specialized milking halls with such milking units as «Parallel», «Carousel» or in stalls with the milking unit «Molokoprovid». The fat and protein mass fraction, dry matter and fat-free dry matter, density, titratable and active acidity, heat resistance and freezing point have been determined according to the accepted techniques. The electrical conductivity of milk has been determined by using the analytical device MD-20 MAS-D-TEC. The total amount of milk bacteria has been determined by reductase reduction test and by seeding method in Petri dish. The milk quality has been investigated by the fermentation and rennet fermentation tests. The higher indices of the fat mass fraction, the protein mass fraction and the dry substance concentration of milk, obtained in specialized milking halls, have been established. This is not statistically significant. Milk, obtained from the milking unit «Molokoprovid», has higher index of titratable acidity, lower thermal stability in comparison with milk, obtained from specialized milking halls with milking units «Parallel» and «Carousel». It has been determined that there is the bacteria insemination increase in milk received from milking cows in stalls in comparison with milk, obtained from milking in specialized halls. Milk, obtained from the milking unit «Carousel», indicates the subclinical form of mastitis in cows or «Carousel» operation violationif there is in 1,8 mS/cm conductivity increase above average index 4,6 mS/cm. Key words: technology, quality and safety of milk, milking, milking unit, milking hall, bacterial insemination.

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