An in situ immunoperoxidase staining procedure for human cell colonies grown in semi-solid agar culture

1986 ◽  
Vol 87 (2) ◽  
pp. 145-148 ◽  
Author(s):  
Rosemary L. Sparrow ◽  
Neil Williams
Keyword(s):  
Human Cell ◽  
Staining Procedure ◽  
Immunoperoxidase Staining ◽  
Agar Culture ◽  
Solid Agar ◽  
Semi Solid

scholarly journals Comparison of Agar and an Agitated, Thin-film, Liquid System for Micropropagation of Ornamental Elephant Ears

HortScience ◽  
2004 ◽  
Vol 39 (5) ◽  
pp. 1088-1092 ◽  
Author(s):  
Jeffrey Adelberg ◽  
Joe Toler
Keyword(s):  
Thin Film ◽  
Growth Medium ◽  
Agar Medium ◽  
Dry Weight ◽  
Liquid System ◽  
Multiplication Rate ◽  
Film Culture ◽  
Agar Culture ◽  
Solid Agar ◽  
Semi Solid

Micropropagation of black-stemmed elephant ear (C. esculenta (L.) Schott `Fontanesii')' and upright elephant ear (A. macrorrhizos G. Don) were compared in semi-solid agar media and agitated, liquid thin-film bioreactor vessels at four explant densities (33, 100, 165, and 330 explants/L of media) using two growth regulator combinations: 1) 1 μm benzylaminopurine (BA)—growth medium, and 2) 3 μm BA plus 3 μm ancymidol—multiplication medium. The thin-film liquid system outperformed agar culture for most measured responses. Some exceptions were relative dry weights at higher explant densities and multiplication rate of Colocasia. When the thin-film liquid system was compared to agar culture, Alocasia explants produced their greatest biomass and had the least residual sugar at the highest explant density. Alocasia explants multiplied most rapidly and had the greatest relative dry weight on liquid media at the low explant densities. Alocasia plants were larger in growth medium than multiplication medium and larger in liquid medium than agar medium. When compared to agar, Colocasia in the thin-film liquid system produced the greatest biomass at the highest explant density in growth medium, had the greatest relative dry weight at the lowest explant density, and used the most sugar at the highest explant density. Alocasia and Colocasia would likely produce greater fresh and dry weight at the highest explant density if additional sugar were supplied during thin-film culture. Greater growth in thin-film culture of Alocasia and Colocasia is due in part, to greater availability of sugar in liquid compared to agar medium.

scholarly journals In Vitro Colony Formation of Carp Granulocytes in a Semi-Solid Agar Culture System.

1993 ◽  
Vol 28 (2) ◽  
pp. 59-63 ◽  
Author(s):  
Tadaaki Moritomo ◽  
Akihide Itou ◽  
Itaru Yamaguchi ◽  
Tasuku Watanabe
Keyword(s):  
Colony Formation ◽  
Culture System ◽  
Agar Culture ◽  
Solid Agar ◽  
Semi Solid

Immunohistochemical identification of lung cells responsive to beta-stimulation with a rise in cAMP

1987 ◽  
Vol 63 (1) ◽  
pp. 434-439 ◽  
Author(s):  
T. B. Casale ◽  
D. Wood ◽  
S. Wescott ◽  
M. Kaliner
Keyword(s):  
Airway Epithelial Cells ◽  
Beta Agonist ◽  
Staining Procedure ◽  
Immunoperoxidase Staining ◽  
Airway Smooth Muscle Cells ◽  
Lung Cells ◽  
Type I ◽  
Airway Epithelial ◽  
Capillary Endothelial Cells

To identify specific lung cells possessing functional beta-adrenergic receptors, we developed an immunoperoxidase-staining procedure capable of in situ localization of cells responding to beta-agonist stimulation with a rise in adenosine 3′,5′-cyclic monophosphate (cAMP). Isoproterenol was instilled into the airways of excised intact guinea pig lungs for 5 min and resulted in a six to eightfold rise in cAMP. Immediately thereafter, the lungs were washed in and fixed with 10% buffered Formalin. Sections were then stained using immunoperoxidase techniques and monoclonal antibodies directed against cAMP. We found that isoproterenol-stimulated lungs had widespread increased staining for immunoreactive cAMP. The specific cells consistently demonstrating marked increases in staining were airway epithelial cells, airway smooth muscle cells, alveolar and parenchymal macrophages, and alveolar lining cells, including both type I and type II cells, and capillary endothelial cells. Of all tissues, the airway epithelium was the most intensely stained area for beta-agonist-induced immunoreactive cAMP. The techniques employed herein should make possible the in situ localization of cells responding to any stimuli capable of increasing cAMP, thereby allowing the specific identification of cells possessing functional adenylate cyclase-linked receptors.

The Effects of Drugs that Cause Neutropenia upon Colony Formation by Bone Marrow Cells in Semi-Solid Agar

1974 ◽  
Vol 46 (5) ◽  
pp. 619-628
Author(s):  
A. Howell ◽  
Susan Chinn ◽  
T. M. Andrews ◽  
R. W. E. Watts
Keyword(s):  
Bone Marrow ◽  
Colony Formation ◽  
Bone Marrow Cells ◽  
Mouse Bone Marrow ◽  
Sodium Aurothiomalate ◽  
Response Curves ◽  
Agar Culture ◽  
Drug Concentrations ◽  
Solid Agar ◽  
Semi Solid

1. The effect of drugs that cause neutropenia has been tested upon normal bone marrow colony formation in semi-solid agar. 2. Thiouracil, methimazole and phenylbutazone do not cause inhibition whereas sodium aurothiomalate, 6-mercaptopurine and chloramphenicol cause dose-related inhibition of colony formation at or below drug concentrations found in serum during therapy. Chloramphenicol also inhibits mouse bone marrow, L cells and Hela cells in agar culture. 3. Dose—response curves have been analysed by the probit method and found to be reproducible.

scholarly journals Routes for salmonella contamination of poultry meat: epidemiological study from hatchery to slaughterhouse

2002 ◽  
Vol 129 (2) ◽  
pp. 253-265 ◽  
Author(s):  
M. HEYNDRICKX ◽  
D. VANDEKERCHOVE ◽  
L. HERMAN ◽  
I. ROLLIER ◽  
K. GRIJSPEERDT ◽  
...  
Keyword(s):  
Horizontal Transmission ◽  
Poultry Meat ◽  
Contamination Level ◽  
Solid Agar ◽  
Determining Factors ◽  
Production Stages ◽  
Animal Material ◽  
Semi Solid ◽  
Therapeutic Doses ◽  
Broiler House

Data were collected on the prevalence of salmonella at different stages during the life cycle of 18 broiler flocks on different farms as well as during slaughter in different poultry slaughterhouses. For the isolation of salmonella, the highest sensitivity (93.9%) was obtained by enrichment in the semi-solid agar Diasalm. The ‘overshoe method’ utilizing several pairs of overshoes provided the highest sensitivity for determining the salmonella status of the broilers during rearing. A clear decrease of the relative importance of the first production stages was demonstrated for the salmonella contamination of the end product, whereas horizontal transmission of salmonella to broilers during rearing and to broiler carcasses in the slaughterhouse was shown to be the main determinative factor. Ten of the 18 flocks received a salmonella positive status with the highest shedding occurring during the first 2 weeks of rearing. The shedding of the animals was significantly negatively influenced by the use of subtherapeutic or therapeutic doses of antibiotics. The intake of portable material in the broiler house was identified as the most important risk factor for horizontal transmission. Significant associations were found between the contamination level of a flock and hygiene of the broiler house, feed and water in the broiler house and both animal and non-animal material sampled in the environment. No correlation was found between contamination during the rearing period and contamination found after slaughtering. The presence of faecal material in the transport crates and predominantly the identity of the slaughterhouse seemed to be the determining factors for carcass quality. Improved hygiene management during transport of broilers and in some slaughterhouses could significantly reduce the risk of salmonella contamination of poultry meat.

scholarly journals Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Robert J. Francis ◽  
Gillian Robb ◽  
Lee McCann ◽  
Bhagwati Khatri ◽  
James Keeble ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Three Dimensional ◽  
Large Field ◽  
Staining Procedure ◽  
3D Analysis ◽  
Mycobacterium Tuberculosis Infection ◽  
In Vivo Models ◽  
Novel Approach

AbstractTuberculosis (TB) preclinical testing relies on in vivo models including the mouse aerosol challenge model. The only method of determining colony morphometrics of TB infection in a tissue in situ is two-dimensional (2D) histopathology. 2D measurements consider heterogeneity within a single observable section but not above and below, which could contain critical information. Here we describe a novel approach, using optical clearing and a novel staining procedure with confocal microscopy and mesoscopy, for three-dimensional (3D) measurement of TB infection within lesions at sub-cellular resolution over a large field of view. We show TB morphometrics can be determined within lesion pathology, and differences in infection with different strains of Mycobacterium tuberculosis. Mesoscopy combined with the novel CUBIC Acid-Fast (CAF) staining procedure enables a quantitative approach to measure TB infection and allows 3D analysis of infection, providing a framework which could be used in the analysis of TB infection in situ.

scholarly journals The influence of physical state on shikimic acid ozonolysis: a case for in situ microspectroscopy

2014 ◽  
Vol 14 (19) ◽  
pp. 10761-10772 ◽  
Author(s):  
S. S. Steimer ◽  
M. Lampimäki ◽  
E. Coz ◽  
G. Grzinic ◽  
M. Ammann
Keyword(s):  
Degradation Rate ◽  
Shikimic Acid ◽  
Bulk Material ◽  
Water Mixture ◽  
Gas Uptake ◽  
X Ray ◽  
Scanning Transmission ◽  
Dry Conditions ◽  
Semi Solid

Abstract. Atmospheric soluble organic aerosol material can become solid or semi-solid. Due to increasing viscosity and decreasing diffusivity, this can impact important processes such as gas uptake and reactivity within aerosols containing such substances. This work explores the dependence of shikimic acid ozonolysis on humidity and thereby viscosity. Shikimic acid, a proxy for oxygenated reactive organic material, reacts with O3 in a Criegee-type reaction. We used an environmental microreactor embedded in a scanning transmission X-ray microscope (STXM) to probe this oxidation process. This technique facilitates in situ measurements with single micron-sized particles and allows to obtain near-edge X-ray absorption fine structure (NEXAFS) spectra with high spatial resolution. Thus, the chemical evolution of the interior of the particles can be followed under reaction conditions. The experiments show that the overall degradation rate of shikimic acid is depending on the relative humidity in a way that is controlled by the decreasing diffusivity of ozone with decreasing humidity. This decreasing diffusivity is most likely linked to the increasing viscosity of the shikimic acid–water mixture. The degradation rate was also depending on particle size, most congruent with a reacto-diffusion limited kinetic case where the reaction progresses only in a shallow layer within the bulk. No gradient in the shikimic acid concentration was observed within the bulk material at any humidity indicating that the diffusivity of shikimic acid is still high enough to allow its equilibration throughout the particles on the timescale of hours at higher humidity and that the thickness of the oxidized layer under dry conditions, where the particles are solid, is beyond the resolution of STXM.

Sign in / Sign up

Export Citation Format

Share Document