Indirect Immunofluorescent Labeling of Viable Cells

2003 ◽  
pp. 261-264
Author(s):  
Robert E. Cunningham
Keyword(s):  
Viable Cells ◽  
Indirect Immunofluorescent

Permeability of boar and bull spermatozoa to the nucleic acid stains propidium iodide or Hoechst 33258, or to eosin: accuracy in the assessment of cell viability

Reproduction ◽  
2000 ◽  
pp. 145-152 ◽  
Author(s):  
B Pintado ◽  
J de la Fuente ◽  
ER Roldan
Keyword(s):  
Nucleic Acid ◽  
Propidium Iodide ◽  
Hoechst 33258 ◽  
Experimental Conditions ◽  
Functional Studies ◽  
Viable Cells ◽  
Direct Microscopic Examination ◽  
Acrosomal Exocytosis ◽  
Viable Sperm ◽  
Boar Spermatozoa

This study was designed to assess whether nucleic acid stains such as propidium iodide and Hoechst 33258 and the cytosolic stain eosin identified equivalent proportions of non-viable cells. Sub-samples of boar spermatozoa stored for up to 72 h, and frozen bull spermatozoa stored in straws and thawed before staining, were exposed to either propidium iodide or Hoechst 33258 alone or in combination. Additional sub-samples were stained with eosin-nigrosin and subsequently with Giemsa. The proportion of non-viable cells identified by propidium iodide alone was equivalent to that observed when it was used in combination with the other fluorescent probe. Similar results were observed for Hoechst 33258. However, direct microscopic examination of sub-samples exposed to both stains revealed that a proportion of spermatozoa stained with propidium iodide did not incorporate Hoechst 33258. This was found consistently in boar and bull spermatozoa under the different experimental conditions used. Quantification showed that the proportion of propidium iodide-positive cells was significantly higher than Hoechst 33258-positive cells. Furthermore, the proportion of propidium iodide-positive cells was higher than cells stained with eosin, but no differences were found between the number of cells stained with Hoechst 33258 or eosin. The proportion of cells stained with propidium iodide was positively correlated with the proportion stained with either Hoechst 33258 or eosin, despite the observation that more cells incorporated propidium iodide. Taken together, these results indicate that there are differences in the ability of fluorescent probes to identify non-viable sperm cells and that this should be considered when staining protocols are used to analyse sperm viability, or when viability is used as a discriminating factor in functional studies, such as those related to acrosomal exocytosis.

Endogenous Viable Cells in Lyopreserved Amnion Retain Differentiation Potential and Anti-Fibrotic Activity In Vitro

2019 ◽  
Author(s):  
Yong Mao ◽  
Tyler Hoffman ◽  
Sandeep Dhall ◽  
Amit Singal ◽  
Malathi Sathyamoorthy ◽  
...  
Keyword(s):  
Differentiation Potential ◽  
Viable Cells

Polylactide-based stent coatings: biodegradable polymeric coatings capable of maintaining sustained release of the thrombolytic enzyme streptokinase

2020 ◽  
Vol 92 (8) ◽  
pp. 1329-1340
Author(s):  
A. G. Kolmakov ◽  
A. S. Baikin ◽  
S. V. Gudkov ◽  
K. N. Belosludtsev ◽  
E. O. Nasakina ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Polymer Films ◽  
De Novo ◽  
Membrane Surface ◽  
Aqueous Media ◽  
Polymeric Coatings ◽  
Short Term ◽  
Thrombolytic Activity ◽  
Viable Cells ◽  
Nitinol Stents

AbstractThe paper describes synthesis and testing of novel biodegradable polylactide-based polymer membranes with desired mechanical properties, which are capable of sustained and directed release of biomacromolecules with high molecular weight (in particular, streptokinase; m.w. 47 kDa). Streptokinase is a pharmaceutical agent, possessing a pronounced thrombolytic activity. The membranes synthesized had a percentage elongation of 2–11% and tensile strength of 25–85 MPa. They were biodegradable – yet being stored in aqueous media in the absence of biological objects, would be dissolved by no more than 10% in 6 months. The synthesized membranes were capable of controlled release of streptokinase into the intercellular space, with the enzyme retaining more than 90% of its initial activity. The rate of streptokinase release from the membranes varied from 0.01 to 0.04 mg/day per cm2 of membrane surface. The membrane samples tested in the work did not have any short-term toxic effects on the cells growing de novo on the membrane surface. The mitotic index of those cells was approximately 1.5%, and the number of non-viable cells on the surface of the polymer films did not exceed 3–4% of their total amount. The implantation of the synthesized polymers – as both individual films and coatings of nitinol stents – was not accompanied by any postoperative complications. The subsequent histological examination revealed no abnormalities. Two months after the implantation of polymer films, only traces of polylactide were found in the implant-surrounding tissues. The implantation of stents coated with streptokinase-containing polymers resulted in the formation of a mature and thick connective-tissue capsules. Thus, the polylactide membranes synthesized and tested in this work are biodegradable, possess the necessary mechanical properties and are capable of sustained and directed release of streptokinase macromolecules.

scholarly journals Bactericidal activity of three different antiseptic ophthalmic preparations as surgical prophylaxis

2021 ◽  
Author(s):  
Daniele Tognetto ◽  
Marco R. Pastore ◽  
Gian Marco Guerin ◽  
Giuliana Decorti ◽  
Martina Franzin ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Povidone Iodine ◽  
Bacterial Load ◽  
Viable Cells ◽  
Polyethylene Glycol 1000 ◽  
Oil Concentration ◽  
Short Time ◽  
First Time ◽  
Antiseptic Solutions

Abstract Purpose In the era of antibiotic resistance, there is an increased interest in antiseptic solutions that might represent a reliable option for ocular surface disinfection. The objective of this study is to compare for the first time three different antiseptic ophthalmic preparations to assess their in vitro antimicrobial activity. Methods The antiseptic activity of three commercial ophthalmic solutions, IODIM (povidone-iodine 0.6% in hyaluronic acid vehicle—Medivis, Catania, Italy), OZODROP (nanoemulsion with ozonated oil—concentration not specified—FBVision, Ophthalmic Pharmaceuticals, Rome, Italy), and DROPSEPT (chlorhexidine 0.02% and vitamin E 0.5% Tocopherol Polyethylene Glycol 1000 Succinate—TPGS, Sooft Italia, Montegiorgio, Italy), was tested in vitro on six reference strains by time-killing assays. Viable cells were evaluated after 1, 15, 30 min; 2, 6, and 24 h exposure by seeding 100 µl of the suspension (or appropriate dilutions) on LB agar or Sabouraud-dextrose agar. All plates were incubated at 37 °C for 24 h and evaluated by manually counting the colonies. Results IODIM solution showed a very rapid microbicidal activity: the number of viable cells for all the tested strains was under the detection limit (less than 10 CFU/ml) already after 1 min exposure, and this result was maintained at every incubation time. The rapid antimicrobial activity of povidone-iodine was not replicated when testing the other two antiseptics. Conclusions The study reports the great efficacy in reducing bacterial load in a very short time of povidone-iodine 0.6% compared with other antiseptic preparations.

CONTINUOUS CULTURE OF BRUCELLA ABORTUS S.19

1961 ◽  
Vol 7 (4) ◽  
pp. 491-505 ◽  
Author(s):  
Andreas H. W. Hauschild ◽  
Hilliard Pivnick
Keyword(s):  
Growth Rate ◽  
Continuous Culture ◽  
Dilution Rate ◽  
Brucella Abortus ◽  
Specific Growth ◽  
Limiting Factor ◽  
Continuous Growth ◽  
Viable Cells ◽  
Metabolic Products ◽  
Growth Of Bacteria

An apparatus is described for the continuous growth of bacteria. Brucella abortus S.19 has been grown in continuous culture for periods up to 3 weeks with populations up to 2 × 1011viable cells per ml and without the establishment of nonsmooth variants.Concentrations between 3 × 109and 2 × 1011cells per ml could be maintained as a function of the dilution rate without the requirement of a known limiting factor in the medium. In a series of steady-state conditions, the specific growth rate increased steadily up to 0.28 hour−1with decreasing population levels.Incidence of mutants was governed by the dilution rate and could also be reduced by various chelating substances.In continuous growth combined with continuous dialysis, population levels were approximately twice those obtained in continuous growth without dialysis. The effect of dialysis appears to be the continuous removal of growth-limiting metabolic products.

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