Protocol for In Situ Detection of Functional Genes of Microorganisms by Two-Pass TSA-FISH

2015 ◽  
pp. 131-144
Author(s):  
Kengo Kubota ◽  
Shuji Kawakami
Keyword(s):  
Functional Genes ◽  
In Situ Detection

Rapid quantification and in situ detection of nitrifying bacteria in biofilms by monoclonal antibody method

2000 ◽  
Vol 41 (4-5) ◽  
pp. 301-308 ◽  
Author(s):  
N. Noda ◽  
H. Ikuta ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fluorescent Antibody ◽  
Nitrifying Bacteria ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Antibody Method ◽  
In Situ Detection ◽  
Rapid Quantification

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.

scholarly journals Single Step In Situ Detection of Surface Protein and MicroRNA in Clustered Extracellular Vesicles Using Flow Cytometry

2021 ◽  
Vol 10 (2) ◽  
pp. 319
Author(s):  
Hee Cheol Yang ◽  
Won Jong Rhee
Keyword(s):  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Molecular Beacon ◽  
Disease Diagnosis ◽  
Single Step ◽  
Flow Cytometer ◽  
Surface Proteins ◽  
Biomarker Detection ◽  
In Situ Detection

Because cancers are heterogeneous, it is evident that multiplexed detection is required to achieve disease diagnosis with high accuracy and specificity. Extracellular vesicles (EVs) have been a subject of great interest as sources of novel biomarkers for cancer liquid biopsy. However, EVs are nano-sized particles that are difficult to handle; thus, it is necessary to develop a method that enables efficient and straightforward EV biomarker detection. In the present study, we developed a method for single step in situ detection of EV surface proteins and inner miRNAs simultaneously using a flow cytometer. CD63 antibody and molecular beacon-21 were investigated for multiplexed biomarker detection in normal and cancer EVs. A phospholipid-polymer-phospholipid conjugate was introduced to induce clustering of the EVs analyzed using nanoparticle tracking analysis, which enhanced the detection signals. As a result, the method could detect and distinguish cancer cell-derived EVs using a flow cytometer. Thus, single step in situ detection of multiple EV biomarkers using a flow cytometer can be applied as a simple, labor- and time-saving, non-invasive liquid biopsy for the diagnosis of various diseases, including cancer.

Real-time in situ detection and source tracing of different soot

Optik ◽  
2021 ◽  
pp. 167711
Author(s):  
Enlai Wan ◽  
Zhongmou Sun ◽  
Yuzhu Liu
Keyword(s):  
Real Time ◽  
Source Tracing ◽  
In Situ Detection

scholarly journals Development of an Easy-to-Operate Underwater Raman System for Deep-Sea Cold Seep and Hydrothermal Vent In Situ Detection

Sensors ◽  
2021 ◽  
Vol 21 (15) ◽  
pp. 5090
Author(s):  
Qingsheng Liu ◽  
Jinjia Guo ◽  
Wangquan Ye ◽  
Kai Cheng ◽  
Fujun Qi ◽  
...  
Keyword(s):  
Deep Sea ◽  
Hydrothermal Vent ◽  
Methane Hydrate ◽  
Sulfur Cycle ◽  
Cold Seep ◽  
Remotely Operated Vehicle ◽  
Bacterial Mats ◽  
Optical Module ◽  
In Situ Detection

As a powerful in situ detection technique, Raman spectroscopy is becoming a popular underwater investigation method, especially in deep-sea research. In this paper, an easy-to-operate underwater Raman system with a compact design and competitive sensitivity is introduced. All the components, including the optical module and the electronic module, were packaged in an L362 × Φ172 mm titanium capsule with a weight of 20 kg in the air (about 12 kg in water). By optimising the laser coupling mode and focusing lens parameters, a competitive sensitivity was achieved with the detection limit of SO42− being 0.7 mmol/L. The first sea trial was carried out with the aid of a 3000 m grade remotely operated vehicle (ROV) “FCV3000” in October 2018. Over 20,000 spectra were captured from the targets interested, including methane hydrate, clamshell in the area of cold seep, and bacterial mats around a hydrothermal vent, with a maximum depth of 1038 m. A Raman peak at 2592 cm−1 was found in the methane hydrate spectra, which revealed the presence of hydrogen sulfide in the seeping gas. In addition, we also found sulfur in the bacterial mats, confirming the involvement of micro-organisms in the sulfur cycle in the hydrothermal field. It is expected that the system can be developed as a universal deep-sea survey and detection equipment in the near future.

scholarly journals Toward Chalcogenide Platform Infrared Sensor Dedicated to the In Situ Detection of Aromatic Hydrocarbons in Natural Waters via an Attenuated Total Reflection Spectroscopy Study

Sensors ◽  
2021 ◽  
Vol 21 (7) ◽  
pp. 2449
Author(s):  
Marion Baillieul ◽  
Emeline Baudet ◽  
Karine Michel ◽  
Jonathan Moreau ◽  
Petr Němec ◽  
...  
Keyword(s):  
Natural Water ◽  
Natural Waters ◽  
Total Reflection ◽  
Attenuated Total Reflection ◽  
Infrared Sensor ◽  
Reflection Spectroscopy ◽  
Infrared Spectral ◽  
Micro Sensor ◽  
In Situ Detection

The objective of this study is to demonstrate the successful functionalization of the surface of a chalcogenide infrared waveguide with the ultimate goal of developing an infrared micro-sensor device. First, a polyisobutylene coating was selected by testing its physico-chemical compatibility with a Ge-Sb-Se selenide surface. To simulate the chalcogenide platform infrared sensor, the detection of benzene, toluene, and ortho-, meta- and para-xylenes was efficaciously performed using a polyisobutylene layer spin-coated on 1 and 2.5 µm co-sputtered selenide films of Ge28Sb12Se60 composition deposited on a zinc selenide prism used for attenuated total reflection spectroscopy. The thickness of the polymer coating was optimized by attenuated total reflection spectroscopy to achieve the highest possible attenuation of water absorption while maintaining the diffusion rate of the pollutant through the polymer film compatible with the targeted in situ analysis. Then, natural water, i.e., groundwater, wastewater, and seawater, was sampled for detection measurement by means of attenuated total reflection spectroscopy. This study is a valuable contribution concerning the functionalization by a hydrophobic polymer compatible with a chalcogenide optical sensor designed to operate in the mid-infrared spectral range to detect in situ organic molecules in natural water.

scholarly journals Mycoplasma hyorhinis as a possible cause of fibrinopurulent meningitis in pigs? - a case series

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Moritz Bünger ◽  
Rene Brunthaler ◽  
Christine Unterweger ◽  
Igor Loncaric ◽  
Maximiliane Dippel ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Clinical Signs ◽  
Case Series ◽  
Upper Respiratory Tract ◽  
Mycoplasma Hyorhinis ◽  
Case Presentation ◽  
Systemic Spread ◽  
In Situ Detection ◽  
On Farm

Abstract Background Mycoplasma hyorhinis is an invader of the upper respiratory tract in swine that is considered to have ubiquitous distribution. It is mainly known for causing polyserositis and polyarthritis in weaned piglets, even though the mechanisms of systemic spread are not fully understood. Mycoplasma hyorhinis has also been associated with other diseases in pigs such as pneumonia or otitis media, but so far has not been known to cause central nervous disorders. This case series reports the isolation of Mycoplasma hyorhinis from cerebrospinal fluid and/ or meningeal swabs from piglets originating from four different piglet producing farms in Austria. Case presentation On farm 1, coughing, stiff movement and central nervous signs occurred in nursery piglets. Mycoplasma hyorhinis was the only pathogen isolated from meningeal swabs from two piglets showing central nervous signs. Fibrinopurulent leptomeningitis was only observed in one piglet. Only one of two nursery piglets from farm 2 showed mild central nervous signs but no histologic lesions; Mycoplasma hyorhinis was isolated from cerebrospinal fluid of the piglet with neurologic signs. Mycoplasma hyorhinis was isolated from cerebrospinal fluid of all three investigated piglets from farm 3, all of which showed central nervous signs and purulent leptomeningitis. Further, Streptococcus suis was isolated from the cerebrospinal fluid of one piglet. Fibrinopurulent leptomeningitis was detected in two piglets from farm 4 that had died overnight without showing any clinical signs and Mycoplasma hyorhinis was isolated from meningeal swabs from both piglets. Conclusion While causality has yet to be proven by experimental infection and in situ detection of the pathogen in histologic sections, the findings of this study and the absence of other pathogens suggest Mycoplasma hyorhinis as a potential causative agent of meningitis in swine.

Precise stepping control of a piezoelectric traveling wave micromotor through embedded in-situ detection

2020 ◽  
Vol 30 (10) ◽  
pp. 105007
Author(s):  
Yicheng Wang ◽  
Tianyu Yang ◽  
Xiaoshi Li ◽  
Jie Cheng ◽  
Yu Chen ◽  
...  
Keyword(s):  
Traveling Wave ◽  
In Situ Detection
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