Chitin distribution in the Oithona digestive and reproductive systems revealed by fluorescence microscopy

10.7287/peerj.preprints.3447v1 ◽

2017 ◽

Cited By ~ 1

Author(s):

Kevin Sugier ◽

Benoit Vacherie ◽

Astrid Cornils ◽

Jean-Louis Jamet ◽

Patrick Wincker ◽

...

Keyword(s):

Fluorescence Microscopy ◽

Comparative Anatomy ◽

Peritrophic Membrane ◽

Internal Anatomy ◽

Reproductive Systems ◽

Marine Food Chain ◽

Oithona Similis ◽

Wide Range ◽

Staining Protocol

Among copepods, which are the most abundant animals on Earth, the genus Oithona is described as one of the most important and plays a major role in the marine food chain and biogeochemical cycles, particularly through the excretion of chitin-coated fecal pellets. Despite the morphology of several Oithona species is well known, knowledge of its internal anatomy and chitin distribution is still limited. To answer this problem, Oithona nana and Oithona similis individuals were stained by WGA-FITC and DAPI for fluorescence microscopy observations. The image analyses allowed a new description of the organization and chitin content of the digestive and reproductive systems of Oithona male and female. Chitin microfibrils were found all along the digestive system from the stomach to the hindgut with a higher concentration at the peritrophic membrane of the anterior midgut. Several midgut shrinkages were observed and proposed to be involved in fecal pellet shaping and motion. Amorphous chitin structures were also found to be a major component of the ducts and seminal vesicles and receptacles. The rapid staining protocol we proposed allowed a new insight into the Oithona internal anatomy and highlighted the role of chitin in the digestion and reproduction. This method could be applied to a wide range of copepods in order to perform comparative anatomy analyses.

Chitin distribution in the Oithona digestive and reproductive systems revealed by fluorescence microscopy

10.7287/peerj.preprints.3447v2 ◽

2018 ◽

Author(s):

Kevin Sugier ◽

Benoit Vacherie ◽

Astrid Cornils ◽

Jean-Louis Jamet ◽

Patrick Wincker ◽

...

Keyword(s):

Fluorescence Microscopy ◽

Comparative Anatomy ◽

Peritrophic Membrane ◽

Internal Anatomy ◽

Reproductive Systems ◽

Marine Food Chain ◽

Oithona Similis ◽

Wide Range ◽

Staining Protocol

Among copepods, which are the most abundant animals on Earth, the genus Oithona is described as one of the most important and plays a major role in the marine food chain and biogeochemical cycles, particularly through the excretion of chitin-coated fecal pellets. Despite the morphology of several Oithona species is well known, knowledge of its internal anatomy and chitin distribution is still limited. To answer this problem, Oithona nana and Oithona similis individuals were stained by WGA-FITC and DAPI for fluorescence microscopy observations. The image analyses allowed a new description of the organization and chitin content of the digestive and reproductive systems of Oithona male and female. Chitin microfibrils were found all along the digestive system from the stomach to the hindgut with a higher concentration at the peritrophic membrane of the anterior midgut. Several midgut shrinkages were observed and proposed to be involved in fecal pellet shaping and motion. Amorphous chitin structures were also found to be a major component of the ducts and seminal vesicles and receptacles. The rapid staining protocol we proposed allowed a new insight into the Oithona internal anatomy and highlighted the role of chitin in the digestion and reproduction. This method could be applied to a wide range of copepods in order to perform comparative anatomy analyses.

Chitin distribution in the Oithona digestive and reproductive systems revealed by fluorescence microscopy

10.7287/peerj.preprints.3447 ◽

2018 ◽

Author(s):

Kevin Sugier ◽

Benoit Vacherie ◽

Astrid Cornils ◽

Jean-Louis Jamet ◽

Patrick Wincker ◽

...

Keyword(s):

Fluorescence Microscopy ◽

Comparative Anatomy ◽

Peritrophic Membrane ◽

Internal Anatomy ◽

Reproductive Systems ◽

Marine Food Chain ◽

Oithona Similis ◽

Wide Range ◽

Staining Protocol

Among copepods, which are the most abundant animals on Earth, the genus Oithona is described as one of the most important and plays a major role in the marine food chain and biogeochemical cycles, particularly through the excretion of chitin-coated fecal pellets. Despite the morphology of several Oithona species is well known, knowledge of its internal anatomy and chitin distribution is still limited. To answer this problem, Oithona nana and Oithona similis individuals were stained by WGA-FITC and DAPI for fluorescence microscopy observations. The image analyses allowed a new description of the organization and chitin content of the digestive and reproductive systems of Oithona male and female. Chitin microfibrils were found all along the digestive system from the stomach to the hindgut with a higher concentration at the peritrophic membrane of the anterior midgut. Several midgut shrinkages were observed and proposed to be involved in fecal pellet shaping and motion. Amorphous chitin structures were also found to be a major component of the ducts and seminal vesicles and receptacles. The rapid staining protocol we proposed allowed a new insight into the Oithona internal anatomy and highlighted the role of chitin in the digestion and reproduction. This method could be applied to a wide range of copepods in order to perform comparative anatomy analyses.

Asbestos Detection with Fluorescence Microscopy Images and Deep Learning

Sensors ◽

10.3390/s21134582 ◽

2021 ◽

Vol 21 (13) ◽

pp. 4582

Author(s):

Changjie Cai ◽

Tomoki Nishimura ◽

Jooyeon Hwang ◽

Xiao-Ming Hu ◽

Akio Kuroda

Keyword(s):

Deep Learning ◽

Fluorescence Microscopy ◽

Occupational Safety ◽

Graphics Processing Unit ◽

Reference Method ◽

Processing Unit ◽

Fiber Concentration ◽

Art Object ◽

Wide Range ◽

Microscopy Images

Fluorescent probes can be used to detect various types of asbestos (serpentine and amphibole groups); however, the fiber counting using our previously developed software was not accurate for samples with low fiber concentration. Machine learning-based techniques (e.g., deep learning) for image analysis, particularly Convolutional Neural Networks (CNN), have been widely applied to many areas. The objectives of this study were to (1) create a database of a wide-range asbestos concentration (0–50 fibers/liter) fluorescence microscopy (FM) images in the laboratory; and (2) determine the applicability of the state-of-the-art object detection CNN model, YOLOv4, to accurately detect asbestos. We captured the fluorescence microscopy images containing asbestos and labeled the individual asbestos in the images. We trained the YOLOv4 model with the labeled images using one GTX 1660 Ti Graphics Processing Unit (GPU). Our results demonstrated the exceptional capacity of the YOLOv4 model to learn the fluorescent asbestos morphologies. The mean average precision at a threshold of 0.5 ([email protected]) was 96.1% ± 0.4%, using the National Institute for Occupational Safety and Health (NIOSH) fiber counting Method 7400 as a reference method. Compared to our previous counting software (Intec/HU), the YOLOv4 achieved higher accuracy (0.997 vs. 0.979), particularly much higher precision (0.898 vs. 0.418), recall (0.898 vs. 0.780) and F-1 score (0.898 vs. 0.544). In addition, the YOLOv4 performed much better for low fiber concentration samples (<15 fibers/liter) compared to Intec/HU. Therefore, the FM method coupled with YOLOv4 is remarkable in detecting asbestos fibers and differentiating them from other non-asbestos particles.

Internalisation of fluorescein isothiocyanate and fluorescein isothiocyanatedextran by suspension-cultured plant cells

Journal of Cell Science ◽

10.1242/jcs.96.4.721 ◽

1990 ◽

Vol 96 (4) ◽

pp. 721-730

Author(s):

LOUISE COLE ◽

JULLIAN COLEMAN ◽

DAVID EVANS ◽

CHRIS HAWES

Keyword(s):

Fluorescence Microscopy ◽

Incubation Period ◽

Degradation Products ◽

Plant Cells ◽

Fluorescein Isothiocyanate ◽

Kinetic Studies ◽

Apparent Difference ◽

Carrot Cells ◽

Intracellular Compartments ◽

Vacuolar System

The uptake of pure non-conjugated fluorescein isothiocyanate (FITC) and of the membraneimpermeant probe FITC—dextran into suspensioncultured carrot cells and protoplasts has been investigated. Commercial samples of a 70K (K=103Mr) FITC—dextran were shown to contain contaminant FITC and/or its degradation products, which were rapidly internalised into the vacuolar system of both cells and protoplasts. However, purified samples of the 70K FITC—dextran were taken up into the vacuoles of cells but not protoplasts after a lh incubation period. This apparent difference in the ability of cells and protoplasts to internalise FITC—dextrans was confirmed using samples of both commercial and purified 20K FITC—dextran as putative endocytotic probes. Both confocal and conventional fluorescence microscopy of FITC—treated cells have shown that FITC was internalised into similar intracellular compartments as was observed in cells treated with three-times purified 70K FITC—dextran. Thus, FITC was a useful fluorophore for rapidly labelling both the putative endocytotic compartments and the pleiomorphic vacuolar system of carrot cells. Kinetic studies indicated that FITC entered the cell by diffusion in the form of the neutral molecule. We have shown that treatment of cells or protoplasts with the drug Probenecid reversibly inhibited the uptake of FITC from the cytoplasm into the vacuole. In addition, the uptake of FITC into isolated vacuoles was enhanced in the presence of Mg-ATP

The internal anatomy of Sandersiella chilenica Stuardo & Vega, 2011 (Crustacea: Cephalocarida)

Journal of Crustacean Biology ◽

10.1093/jcbiol/ruz091 ◽

2019 ◽

Vol 40 (2) ◽

pp. 176-180

Author(s):

Claudio Valdovinos-Zarges

Keyword(s):

Light Microscopy ◽

Internal Anatomy ◽

Reproductive Systems ◽

Central Chile ◽

Type Series ◽

Histological Sections ◽

Organ Systems ◽

Soft Bottoms

Abstract The gross internal anatomy of the cephalocarid crustacean Sandersiella chilenicaStuardo & Vega, 2011 is described based on light microscopy of histological sections of three specimens of the type series, all collected in coastal soft bottoms off central Chile. Observations mainly concern the nervous, digestive, and reproductive systems, heart, haemocoel, and skeleto-musculature. The main organ systems and musculature are similar to those previously described for Hutchinsoniella macracanthaSanders, 1955, Lightiella magdaleninaCarcupino, Floris, Addis, Castelli & Curini-Galletti, 2006, and Sandersiella sp.

Quantification of Platelet Adsesion in vivo

10.1055/s-0039-1687604 ◽

1979 ◽

Cited By ~ 1

Author(s):

H. Kortenhaus ◽

G.V.R. Bom

Keyword(s):

Fluorescence Microscopy ◽

Small Proportion ◽

Platelet Rich Plasma ◽

Fluorescein Isothiocyanate ◽

Cheek Pouch ◽

Golden Hamsters ◽

Cardiac Blood ◽

Acid Citrate Dextrose ◽

Citrate Dextrose

Golden hamsters (80-100g) were anaesthetised with pentobarbital and cardiac blood was collected into acid-citrate dextrose. Platelet-rich plasma brought to pH 6.2 was incubated at 37° with fluorescein-isothiocyanate (FITC: 100 μg/ml) for 10 min (Kortenhaus , Webelmann and Schroer: to be published). The platelets were injected i.v. into another hamster and observed in the cheek pouch circulation by fluorescence microscopy at x 500.In venules a small proportion of platelets were arrested, most for less than one sec., about 20% for up to 2 min and about 3% for longer. There was no correlation with rolling granulocyte frequencies (A therton & Born, 1972, Journal of Fhysiology, 222, 447).

Nur77 limits endothelial barrier disruption to LPS in the mouse lung

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00425.2018 ◽

2019 ◽

Vol 317 (5) ◽

pp. L615-L624 ◽

Cited By ~ 1

Author(s):

Ni Zhu ◽

Guan-Xin Zhang ◽

Bing Yi ◽

Zhi-Fu Guo ◽

Soohwa Jang ◽

...

Keyword(s):

Cultured Cells ◽

Serum Proteins ◽

Fluorescein Isothiocyanate ◽

Lavage Fluid ◽

Endothelial Barrier ◽

Pulmonary Vasculature ◽

Mouse Lung ◽

Orphan Nuclear Receptor ◽

Protective Effects ◽

Wide Range

Nur77 is an orphan nuclear receptor implicated in the regulation of a wide range of biological processes, including the maintenance of systemic blood vessel homeostasis. Although Nur77 is known to be expressed in the lung, its role in regulating pulmonary vascular functions remains entirely unknown. In this study, we found that Nur77 is expressed at high levels in the lung, and its expression is markedly upregulated in response to LPS administration. While the pulmonary vasculature of mice that lacked Nur77 appeared to function normally under homeostatic conditions, we observed a dramatic decrease in its barrier functions after exposure to LPS, as demonstrated by an increase in serum proteins in the bronchoalveolar lavage fluid and a reduction in the expression of endothelial junctional proteins, such as vascular endothelial cadherin (VE-cadherin) and β-catenin. Similarly, we found that siRNA knockdown of Nur77 in lung microvascular endothelial cells also reduced VE-cadherin and β-catenin expression and increased the quantity of fluorescein isothiocyanate-labeled dextran transporting across LPS-injured endothelial monolayers. Consistent with Nur77 playing a vascular protective role, we found that adenoviral-mediated overexpression of Nur77 both enhanced expression of VE-cadherin and β-catenin and augmented endothelial barrier protection to LPS in cultured cells. Mechanistically, Nur77 appeared to mediate its protective effects, at least in part, by binding to β-catenin and preventing its degradation. Our findings demonstrate a key role for Nur77 in the maintenance of lung endothelial barrier protection to LPS and suggest that therapeutic strategies aimed at augmenting Nur77 levels might be effective in treating a wide variety of inflammatory vascular diseases of the lung.

Mounting Media and Antifade Reagents

Microscopy Today ◽

10.1017/s1551929500055176 ◽

2006 ◽

Vol 14 (1) ◽

pp. 34-39

Author(s):

Tony J. Collins

Keyword(s):

Refractive Index ◽

Fluorescence Microscopy ◽

Fluorescence Signal ◽

Signal Loss ◽

Biomedical Sciences ◽

Optical Aberrations ◽

Wide Range

In the biomedical sciences, samples are mounted in a wide variety of media for examination by microscope. There are a wide variety of mounting media available with a correspondingly wide range of properties. Using the incorrect mounting medium may cause signal loss and optical aberrations; the correct mounting medium avoids such aberrations and preserves fluorescence signal with “anti-fading” properties. This article introduces mounting media for fluorescence microscopy, providing descriptions of their constituents and their properties, as well as accounts of users' experienceMore detailed reviews of antifade reagents have been published by Ono et al. and Longin et al.. Papers describing the effect of refractive index (RI) mismatch have been published by Diaspro et al. and Hell et al..

Distribution of chitin in the yeast cell wall. An ultrastructural and chemical study.

The Journal of Cell Biology ◽

10.1083/jcb.85.2.199 ◽

1980 ◽

Vol 85 (2) ◽

pp. 199-212 ◽

Cited By ~ 86

Author(s):

J Molano ◽

B Bowers ◽

E Cabib

Keyword(s):

Cell Wall ◽

Fluorescence Microscopy ◽

Lectin Binding ◽

Strong Association ◽

Fluorescein Isothiocyanate ◽

Chemical Study ◽

Total Radioactivity ◽

Lytic Enzymes ◽

Derivatives Of ◽

Lateral Walls

The distribution of chitin in Saccharomyces cervisiae primary septa and cell walls was studied with three methods: electron microscopy of colloidal gold particles coated either with wheat germ agglutinin or with one of two different chitinases, fluorescence microscopy with fluorescein isothiocyanate derivatives of the same markers, and enzymatic treatments of [14C]glucosamine-labeled cells. The septa were uniformly and heavily labeled with the gold-attached markers, an indication that chitin was evenly distributed throughout. To study the localization of chitin in lateral walls, alkali-extracted cell ghosts were used. Observations by electron and fluorescence microscopy suggest that lectin-binding material is uniformly distributed over the whole cell ghost wall. This material also appears to be chitin, on the basis of the analysis of the products obtained after treatment of 14C-labeled cell ghosts with lytic enzymes. The chitin of lateral walls can be specifically removed by treatment with beta-(1 leads to 6)-glucanase containing a slight amount of chitinase. During this incubation approximately 7% of the total radioactivity is solubilized, about the same amount liberated when lateral walls of cell ghosts are completely digested with snail glucanase yield primary septa. It is concluded that the remaining chitin, i.e., greater than 90% of the total, is in the septa. The facilitation of chitin removal from the cell wall by beta-(1 leads to 6)-glucanase indicates a strong association between chitin and beta-(1 leads to 6)-glucan. Covalent linkages between the two polysaccharides were not detected but cannot be excluded.