scholarly journals Differentiation of Methanosaeta concilii andMethanosarcina barkeri in Anaerobic Mesophilic Granular Sludge by Fluorescent In Situ Hybridization and Confocal Scanning Laser Microscopy

1999 ◽  
Vol 65 (5) ◽  
pp. 2222-2229 ◽  
Author(s):  
Sylvie Rocheleau ◽  
Charles W. Greer ◽  
John R. Lawrence ◽  
Christiane Cantin ◽  
Louise Laramée ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Outer Layer ◽  
Fluorescent In Situ Hybridization ◽  
Methanosarcina Barkeri ◽  
Confocal Scanning Laser Microscopy ◽  
Oligonucleotide Probes ◽  
Scanning Laser Microscopy ◽  
Confocal Scanning ◽  
Confocal Scanning Laser

ABSTRACT Oligonucleotide probes, designed from genes coding for 16S rRNA, were developed to differentiate Methanosaeta concilii, Methanosarcina barkeri, and mesophilic methanogens. All M. concilii oligonucleotide probes (designated MS1, MS2, and MS5) hybridized specifically with the target DNA, but MS5 was the most specific M. concilii oligonucleotide probe.Methanosarcina barkeri oligonucleotide probes (designated MB1, MB3, and MB4) hybridized with different Methanosarcinaspecies. The MB4 probe specifically detected Methanosarcina barkeri, and the MB3 probe detected the presence of all mesophilic Methanosarcina species. These new oligonucleotide probes facilitated the identification, localization, and quantification of the specific relative abundance of M. concilii and Methanosarcina barkeri, which play important roles in methanogenesis. The combined use of fluorescent in situ hybridization with confocal scanning laser microscopy demonstrated that anaerobic granule topography depends on granule origin and feeding. Protein-fed granules showed no layered structure with a random distribution of M. concilii. In contrast, a layered structure developed in methanol-enriched granules, where M. barkeri growth was induced in an outer layer. This outer layer was followed by a layer composed of M. concilii, with an inner core of M. concilii and other bacteria.

scholarly journals Direct In Situ Viability Assessment of Bacteria in Probiotic Dairy Products Using Viability Staining in Conjunction with Confocal Scanning Laser Microscopy

2001 ◽  
Vol 67 (1) ◽  
pp. 420-425 ◽  
Author(s):  
M. A. E. Auty ◽  
G. E. Gardiner ◽  
S. J. McBrearty ◽  
E. O. O'Sullivan ◽  
D. M. Mulvihill ◽  
...  
Keyword(s):  
Milk Powder ◽  
Scanning Laser ◽  
Confocal Scanning Laser Microscopy ◽  
Laser Microscopy ◽  
Viability Staining ◽  
Scanning Laser Microscopy ◽  
Confocal Scanning ◽  
Confocal Scanning Laser ◽  
Light Staining

ABSTRACT The viability of the human probiotic strains Lactobacillus paracasei NFBC 338 and Bifidobacterium sp. strain UCC 35612 in reconstituted skim milk was assessed by confocal scanning laser microscopy using the LIVE/DEAD BacLight viability stain. The technique was rapid (<30 min) and clearly differentiated live from heat-killed bacteria. The microscopic enumeration of various proportions of viable to heat-killed bacteria was then compared with conventional plating on nutrient agar. Direct microscopic enumeration of bacteria indicated that plate counting led to an underestimation of bacterial numbers, which was most likely related to clumping. Similarly, LIVE/DEAD BacLight staining yielded bacterial counts that were higher than cell numbers obtained by plate counting (CFU) in milk and fermented milk. These results indicate the value of the microscopic approach for rapid viability testing of such probiotic products. In contrast, the numbers obtained by direct microscopic counting for Cheddar cheese and spray-dried probiotic milk powder were lower than those obtained by plate counting. These results highlight the limitations of LIVE/DEAD BacLight staining and the need to optimize the technique for different strain-product combinations. The minimum detection limit for in situ viability staining in conjunction with confocal scanning laser microscopy enumeration was ∼108 bacteria/ml (equivalent to ∼107 CFU/ml), based on Bifidobacterium sp. strain UCC 35612 counts in maximum-recovery diluent.

In Situ Observation of Inclusions Behavior in Molten State and Solidifying Interface of Tire Cord Steel

2014 ◽  
Vol 881-883 ◽  
pp. 1584-1587 ◽  
Author(s):  
Zhi Liang Zhu ◽  
Guo Jun Ma ◽  
Chen Fan Yu
Keyword(s):  
Solidification Front ◽  
Confocal Scanning Laser Microscopy ◽  
Molten State ◽  
Attraction Force ◽  
Tire Cord ◽  
Scanning Laser Microscopy ◽  
Tire Cord Steel ◽  
Confocal Scanning ◽  
Confocal Scanning Laser

In this study, a confocal scanning laser microscopy was used to in situ observe the collision and aggregation behavior of inclusions in molten state and the pushing/engulfing behavior in the solidifying interface of tire cord steel. The results show that the inclusion particles will be captured into the grains in the solidification front for the inclusions with a diameter are larger than 53.4μm at a cooling rate of 1 K/s. Moreover, the attraction force of inclusion particles in molten state is in the order of 10-16N~10-14 N in tire cord steel.

In Situ Localization of Escherichia coli O157:H7 in Food by Confocal Scanning Laser Microscopy

2005 ◽  
Vol 68 (3) ◽  
pp. 482-486 ◽  
Author(s):  
MARK AUTY ◽  
GERALDINE DUFFY ◽  
DAVID O'BEIRNE ◽  
AOIFE MCGOVERN ◽  
ELAINE GLEESON ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Scanning Laser ◽  
Confocal Scanning Laser Microscopy ◽  
Escherichia Coli O157 ◽  
Laser Microscopy ◽  
E Coli ◽  
Scanning Laser Microscopy ◽  
Confocal Scanning ◽  
Confocal Scanning Laser

The aim of this study was to use confocal scanning laser microscopy to examine the in situ localization of Escherichia coli O157:H7 on beef (knuckle or brisket) and carrots and in semisoft cheese made from pasteurized milk. Using a combination of specific immunolabeling and dual-excitation confocal scanning laser microscopy, it was possible to clearly demonstrate the localization of E. coli O157:H7 within various food types. In carrots, bacteria were found mainly at cell junctions and in intracellular spaces up to 50 μm deep. In beef, bacteria were located primarily between muscle fibers and within connective tissue (at a depth of 25 μm), whereas in cheese the bacteria occurred singly or in small clumps of up to 10 cells and were observed within the protein matrix of the cheese. These results revealed how E. coli O157:H7 can penetrate beef and carrot surfaces where it is protected from decontamination processes.

Computer-Assisted Analysis of Multi-Color Confocal Microscopic Datasets: Automated Light Microscopic Discrimination of Synaptic Relationships

1996 ◽  
Vol 54 ◽  
pp. 284-285
Author(s):  
M Wessendorf ◽  
A Beuning ◽  
D Cameron ◽  
J Williams ◽  
C Knox
Keyword(s):  
Nerve Fibers ◽  
Confocal Scanning Laser Microscopy ◽  
Computer Assisted ◽  
Nerve Terminals ◽  
Neuronal Somata ◽  
Scanning Laser Microscopy ◽  
Confocal Scanning ◽  
Entire Cell ◽  
Confocal Scanning Laser ◽  
Assisted Analysis

Multi-color confocal scanning-laser microscopy (CSLM) allows examination of the relationships between neuronal somata and the nerve fibers surrounding them at sub-micron resolution in x,y, and z. Given these properties, it should be possible to use multi-color CSLM to identify relationships that might be synapses and eliminate those that are clearly too distant to be synapses. In previous studies of this type, pairs of images (e.g., red and green images for tissue stained with rhodamine and fluorescein) have been merged and examined for nerve terminals that appose a stained cell (see, for instance, Mason et al.). The above method suffers from two disadvantages, though. First, although it is possible to recognize appositions in which the varicosity abuts the cell in the x or y axes, it is more difficult to recognize them if the apposition is oriented at all in the z-axis—e.g., if the varicosity lies above or below the neuron rather than next to it. Second, using this method to identify potential appositions over an entire cell is time-consuming and tedious.

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