“Sporotan” a new fluorescent stain for identifying cryptic spores of Rhodobacter johrii

2020 ◽  
Vol 177 ◽  
pp. 106019
Author(s):  
Ashif Ali ◽  
Kumar Gaurav ◽  
N. Senthilnathan ◽  
T.P. Radhakrishnan ◽  
Sasikala Ch ◽  
...  
Keyword(s):  
Fluorescent Stain

scholarly journals A microscopic approach to investigate bacteria under in situ conditions in sea-ice samples

2001 ◽  
Vol 33 ◽  
pp. 304-310 ◽  
Author(s):  
Karen Junge ◽  
Christopher Krembs ◽  
Jody Deming ◽  
Aaron Stierle ◽  
Hajo Eicken
Keyword(s):  
Sea Ice ◽  
Pore Space ◽  
Three Dimensional ◽  
Epifluorescence Microscopy ◽  
Microbial Populations ◽  
Microbial Life ◽  
Dimensional Network ◽  
In Situ Conditions ◽  
Fluorescent Stain

AbstractMicrobial populations and activity within sea ice have been well described based on bulk measurements from melted sea-ice samples. However, melting destroys the micro-environments within the ice matrix and does not allow for examination of microbial populations at a spatial scale relevant to the organism. Here, we describe the development of a new method allowing for microscopic observations of bacteria localized within the three-dimensional network of brine inclusions in sea ice under in situ conditions. Conventional bacterial staining procedures, using the DNA-specific fluorescent stain DAPI, epifluorescence microscopy and image analysis, were adapted to examine bacteria and their associations with various surfaces within microtomed sections of sea ice at temperatures from −2° to −15°C. The utility and sensitivity of the method were demonstrated by analyzing artificial sea-ice preparations of decimal dilutions of a known bacterial culture. When applied to natural, particle-rich sea ice, the method allowed distinction between bacteria and particles at high magnification. At lower magnifications, observations of bacteria could be combined with those of other organisms and with morphology and particle content of the pore space. The method described here may ultimately aid in discerning constraints on microbial life at extremely low temperatures.

scholarly journals Glucosylceramide Plays a Role in Fungal Germination, Lipid Raft Organization and Biofilm Adhesion of the Pathogenic Fungus Scedosporium aurantiacum

2020 ◽  
Vol 6 (4) ◽  
pp. 345
Author(s):  
Victor Pereira Rochetti ◽  
Rodrigo Rollin-Pinheiro ◽  
Evely Bertulino de Oliveira ◽  
Mariana Ingrid Dutra da Silva Xisto ◽  
Eliana Barreto-Bergter
Keyword(s):  
Cell Surface ◽  
Pathogenic Fungus ◽  
Clinical Manifestations ◽  
Tube Length ◽  
Total Biomass ◽  
Fungal Cell ◽  
Germ Tube Length ◽  
Wide Range ◽  
Fluorescent Stain ◽  
Altered Lipid

Infections caused by Scedosporium species present a wide range of clinical manifestations, from superficial to disseminated, especially in immunocompromised patients. Glucosylceramides (GlcCer) are glycosphingolipids found on the fungal cell surface and play an important role in growth and pathogenicity processes in different fungi. The present study aimed to evaluate the structure of GlcCer and its role during growth in two S. aurantiacum isolates. Purified GlcCer from both isolates were obtained and its chemical structure identified by mass spectrometry. Using ELISA and immunofluorescence techniques it was observed that germination and NaOH-treatment of conidia favor GlcCer exposure. Monoclonal anti-GlcCer antibody reduced germination when cultivated with the inhibitor of melanin synthesis tricyclazole and also reduced germ tube length of conidia, both cultivated or not with tricyclazole. It was also demonstrated that anti-GlcCer altered lipid rafts organization, as shown by using the fluorescent stain filipin, but did not affect the susceptibility of the cell surface to damaging agents. Anti-GlcCer reduced total biomass and viability in biofilms formed on polystyrene plates. In the presence of anti-GlcCer, germinated S. aurantiacum conidia and biofilms could not adhere to polystyrene with the same efficacy as control cells. These results highlight the relevance of GlcCer in growth processes of S. aurantiacum.

scholarly journals Tuberculous mycobacteria bacilli fluorescence and compare with Ziehl- Neelsen stain in fine-needle aspiration cytology of tubercular lymphnode

2016 ◽  
Vol 2 (2) ◽  
pp. 66
Author(s):  
Mani Krishna ◽  
Adesh Kumar
Keyword(s):  
Infectious Disease ◽  
Mycobacterium Tuberculosis ◽  
Fine Needle Aspiration ◽  
Fine Needle Aspiration Cytology ◽  
Needle Aspiration ◽  
Aspiration Cytology ◽  
Fine Needle ◽  
Fluorescent Method ◽  
Fluorescent Stain ◽  
Needle Aspiration Cytology

<p class="abstract"><strong>Background:</strong> Tuberculosis is infectious disease caused by <em>Mycobacterium tuberculosis</em>. There are various methods for diagnosis of tuberculosis such as direct clinical material examination of tubercular bacilli by Ziehl – Neelsen (ZN) staining, demonstration of tubercular bacilli by auramine – rhodamine (AR) staining and autofluorescence (AF).</p><p class="abstract"><strong>Methods:</strong> Present study was done clinically suspected tubercular patients. All received samples Zn stain, fluorescent stain and PAP stain were applied.</p><p class="abstract"><strong>Results:</strong> Among the clinically suspected patients 88 was diagnosed with tuberculosis. Female preponderance was noted accounting for 60.23% (53/88) of cases. Of the 88 aspirates, the smear positivity for acid fast bacilli (AFB) on the ZN method was 37.5% (33/88) while the positivity increased to 81.82% (72/88) on the AR fluorescent method and 86.36% (76/88) on AF.</p><strong>Conclusions:</strong> AF staining is more sensitive than the auramine – rhodamine fluorescent and ZN staining in demonstration of mycobacterium bacilli in fine needle aspiration cytology of tubercular lymphnode.

Fine structure of the kidneys of osmotically stressed Mytilus, Mercenari, and Anodonta

1986 ◽  
Vol 64 (12) ◽  
pp. 2779-2787 ◽  
Author(s):  
Hamidur R. Khan ◽  
Mary Lou Ashton ◽  
A. S. M. Saleuddin
Keyword(s):  
Basal Membrane ◽  
Ultrastructural Changes ◽  
Freshwater Bivalve ◽  
Septate Junctions ◽  
Intramembrane Particle ◽  
Marine Bivalves ◽  
Fluorescent Stain ◽  
Cytoskeletal Elements ◽  
Lateral Cell ◽  
Membrane Infoldings

Osmotically induced ultrastructural changes in the kidneys of the freshwater bivalve Anodonta and the marine bivalves Mytilus and Mercenaria were studied. Osmotic stresses were given to Anodonta by keeping them in distilled water or in 6% seawater, and to Mytilus and Mercenaria by keeping them in 50% seawater for various periods. In all of these bivalves, the convoluted, single cell layered kidney epithelia displayed wide lateral intercellular spaces as well as extracellular spaces in the basal membrane infoldings during hyposmotic stress. These spaces were greatly reduced when the animals were kept in isosmotic media (i.e., isosmotic to their respective hemolymphs). The kidney cells contained abundant cytoskeletal elements and microfilaments were often observed in bundles in the basal membrane infoldings. Actin was observed in the basal membrane infoldings using the specific fluorescent stain nitrobenzoxadiazole-phallacidin. The cell contacts of the kidney epthelia were studied in platinum replicas of freeze-fractured tissues. The lateral cell membrane and basal membrane infoldings contained many gap junctions. Many rows of dense intramembrane particles of septate junctions were observed in the kidneys of animals from isosmotic media. The septate junctions in the kidneys of aminals from hyposmotic media contained either fewer intramembrane particle rows or many sinuous intramembrane particle rows. The site of prourine formation in mollusks are discussed.

301 THE EFFECT OF EQUINE FOLICULLAR FLUID ON IN VITRO INDUCTION OF THE ACROSOME REACTION IN FROZEN - THAWED STALLION SPERMATOZOA

2010 ◽  
Vol 22 (1) ◽  
pp. 307
Author(s):  
D. S. Silva ◽  
P. Rodriguez ◽  
N. S. Arruda ◽  
R. Rodrigues ◽  
J. L. Rodrigues
Keyword(s):  
Contact Area ◽  
Follicular Fluid ◽  
Acrosome Reaction ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Heparin Group ◽  
Fluorescent Stain ◽  
In Vitro Induction

The capacitation process occurs in vivo upon exposure of the spermatozoa through the female reproductive tract, but can be induced in vitro in the presence of several compounds. This study was conducted to assess the effect of heparin or equine follicular fluid on hyperactivated motility and in vitro induction acrosome reaction swim-up method with frozen-thawed stallion semen. Two hundred microliters of frozen-thawed equine semen was placed in a tube (45°C) to increase contact area and incubated at 37°C for 1 h. After incubation 800 μL of the supernatant was collected by centrifugation (500 × g, 10 min) to collect spermatozoa. The resulting pellet was resuspended in capacitation medium Fert-TALP supplemented with 5.0 μg mL-1 heparin or 100% follicular fluid and incubated for different times (1, 2, 3, 4, and 5 h) at 37°C. After incubation the hyperactivated motility and acrosome-reacted spermatozoa were evaluated. Hoechst stain was used to differentiate live and dead spermatozoa, and chlortetracycline (CTC) fluorescent stain was used to assess the capacitation response of sperm; data were analyzed by ANOVA. The effect of equine follicular fluid resulted in improved percentage of spermatozoa with acrosome reaction at all times of incubation (60, 63, 57, 52, and 58%) but immediately after 3 h of incubation, the hyperactivated motility decreased in heparin group and follicular fluid (42 and 30%, respectively).

Effects of chronic exposure to 2G/3G cell phone radiationon in vitro maturation of bovine oocytes

2016 ◽  
Author(s):  
Yusuf Ziya Guzey ◽  
Ali Galip Onal
Keyword(s):  
Treatment Group ◽  
Mobile Phone ◽  
Cell Phone ◽  
In Vitro Maturation ◽  
Bovine Oocyte ◽  
Maturation Period ◽  
Mobile Phone Use ◽  
Fluorescent Stain ◽  
Bovine Oocytes

Mobile phone use has arisen and rapidly been increased during last decades and therefore electromagnetic radiation or heat sourcing from mobile phones and its possible damages has become focus of interest. The aim of the present study was to investigate the effects of mobile phone signals on in vitro maturation of bovine oocytes. A total of 224 bovine cumulus oocyte complexes were matured in vitro and separated into two groups, and only treatment group received mobile phone signals for 100 min by calling 5 min/h. Following maturation period, oocytes were stained by fluorescent stain to assess maturation rates. The results showed that oocytes reaching Metaphase I and Metaphase II stage were 35.2±2.24 % and 48.2±2.21 % for treatment group whereas 11.9±1.45 % and 73.8±4.60 % for controls, respectively (P<0.01). In conclusion, bovine oocyte maturation has been significantly reduced by mobile phone signals.

An evaluation of rhodamine-labeled lysozyme as a fluorescent stain for in situ soil bacteria

1972 ◽  
Vol 18 (6) ◽  
pp. 933-936 ◽  
Author(s):  
L. J. McElroy ◽  
L. E. Casida Jr.
Keyword(s):  
Fluorescence Microscopy ◽  
Soil Bacteria ◽  
Soil Microorganisms ◽  
Fluorescein Isothiocyanate ◽  
Soil Microflora ◽  
Light Diffraction ◽  
Enzyme Conjugate ◽  
Fluorescent Stain ◽  
Diffraction Microscopy

Rhodamine-labeled lysozyme, in conjunction with fluorescein isothiocyanate conjugated to gelatin as a counterstain, was evaluated as a fluorescent stain for resident soil microorganisms. Preparations were observed by light-diffraction microscopy, transmitted incandescent-light microscopy, and reflected ultraviolet-fluorescence microscopy using a modified light-diffraction microscope, and by conventional transmitted incandescent and ultraviolet-fluorescence microscopy. Laboratory-grown microbial cultures superimposed on the resident soil microflora, as well as the non-dormant resident soil microorganisms, usually fluoresced properly when the stain and counterstain were applied to soil, and the blocking test with non-conjugated lysozyme was effective. What were assumed to be semidormant cells fluoresced to a lesser degree, and the blocking test was only partially effective. In contrast, the dormant resident soil microorganisms usually did not become stained by the methods used. A possible explanation is that a peripheral structural component surrounding many of the resident dormant cells in soil may have rendered their cell-wall substrate inaccessible to the enzyme conjugate.

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