The Experts below are selected from a list of 291 Experts worldwide ranked by ideXlab platform
Urs Wyss - One of the best experts on this subject based on the ideXlab platform.
-
In vivo observations of the arbuscular mycorrhizal fungus Glomus mosseae in roots by confocal laser scanning microscopy
Fungal Biology, 1999Co-Authors: Horst Vierheiling, Christine Juge, Annette Böckenhoff, Michael Knoblauch, Florian M W Grundler, Yves Piche, Urs WyssAbstract:Confocal laser scanning microscopy was used for in vivo observations of autofluorescent Fungal Structures of Glomus mosseae. in intact, living rye-grass roots. Clear images of hyphae and collapsed arbuscules could be obtained without staining or sectioning roots. As rye-grass roots are highly transparent, with only a few cell layers above the vascular cylinder, they provide an excellent in vivo system for microscopical confocal laser scanning studies of arbuscular mycorrhizal Structures.
-
ink and vinegar a simple staining technique for arbuscular mycorrhizal fungi
Applied and Environmental Microbiology, 1998Co-Authors: Horst Vierheilig, Urs Wyss, Andrew P Coughlan, Yves PicheAbstract:We developed a reliable, inexpensive, and simple method for staining arbuscular-mycorrhizal Fungal colonizations in root tissues. Apart from applications in research, this nontoxic, high-quality staining method also could be of great utility in teaching exercises. After adequate clearing with KOH, an ink-vinegar solution successfully stained all Fungal Structures, rendering them clearly visible.
S. Dickson - One of the best experts on this subject based on the ideXlab platform.
-
Visualisation of mycorrhizal Fungal Structures and quantification of their surface area and volume using laser scanning confocal microscopy
Mycorrhiza, 1999Co-Authors: S. Dickson, P. KolesikAbstract:A method has been developed for the visualisation and three-dimensional (3D) measurement of mycorrhizal Fungal Structures inside plant roots. Sections of Allium porrum L. roots colonised by Glomus sp. 'City Beach' (WUM 16) and Lilium sp. roots colonised by Scutellospora calospora (Nicol. & Gerd.) Walker & Sanders (WUM 12(2)) were stained with acid fuchsin. This allowed fluorescence from the Fungal Structures to be observed under a laser scanning confocal microscope (LSCM) without interference from the plant cells. A series of horizontal optical sections were collected from a Glomus sp. arbuscule and from a hyphal coil of S. calospora. These data were used to produce extended focus images. Axial distortion in microscopic visualisation due to the refractive index mismatch between the immersion and mounting media was quantified using vertical scanning of the hyphae. A correction factor of 0.71 μm was used for the z-interval between the xy-slices. A series of binary xy-images from each structure was rendered into a 3D graphical model for viewing. The volume and surface area of the Structures were estimated using computerised 3D measurement and also by stereological integration of binary xy-images. With both Structures, the surface area estimates varied greatly between the two measuring systems, whereas differences in volume estimates were small. Computerised 3D measurement was considered more accurate than stereological integration of confocal binary images.
-
Evaluation of Vesicular-Arbuscular Mycorrhizal Colonisation by Staining
Mycorrhiza Manual, 1998Co-Authors: S. Dickson, S. E. SmithAbstract:VA mycorrhizal plant roots contain a range of Fungal Structures such as external and internal hyphae, arbuscules, vesicles, and sometimes hyphal coils. These Structures may be related to function. For example, arbuscules are considered the major site of mineral nutrient transfer, while intercellular hyphae have been proposed to be responsible for carbohydrate transfer (see Smith and Smith 1990; Gianinazzi-Pearson et al. 1991). So, in order to evaluate VA mycorrhizal colonisation in roots, methods of staining need to be able to distinguish the different Structures formed by the fungus. The diversity of Fungal Structures and their quantity may lead to several choices in how the degree of colonisation within the root should be determined to distinguish physiologically active fungus. Different types of investigation may also require specialised methods such as vital staining. This aspect has often been overlooked, but is crucial for physiological, developmental and biodiversity studies.
-
VISUALIZATION OF MYCORRHIZAL Fungal Structures IN RESIN EMBEDDED TISSUES WITH XANTHENE DYES USING LASER SCANNING CONFOCAL MICROSCOPY
Canadian Journal of Botany, 1998Co-Authors: L. H. Melville, S. Dickson, Melissa L. Farquhar, Sally E. Smith, R. Larry PetersonAbstract:The xanthene dyes sulforhordamine G, phloxine B, rose Bengal, and 4,5,6,7-tetrachloroflorescein were used as fluorochromes for laser scanning confocal microscopy of LR-White resin-embedded mycorrhizae. Sulforhodamine G was the most effective dye, giving an even staining of cell components throughout the material, with minimal background fluorescence of LR-White resin. Confocal microscopy of stained blocks of tissue on a slide, viewed without the use of a coverslip, revealed the three-dimensional nature of various mycorrhizal Structures; these Structures included arbuscules, vesicles, and coiled hyphae in arbuscular mycorrhizae; coiled hyphae in orchid mycorrhizae; mantle and Hartig net hyphae in ectomycorrhizae; and intracellular hyphae in arbutoid mycorrhizae. Sections mounted on slides viewed with confocal microscopy provided exceptional clarity of Fungal form and cytoplasmic contents and showed the relationship to the plant cells, also with negligible background fluorescence. Mounting and staining bloc...
Cheng Huang - One of the best experts on this subject based on the ideXlab platform.
-
Frozen in time: a new method using cryo-scanning electron microscopy to visualize root-Fungal interactions.
The New phytologist, 2006Co-Authors: Stephen Johnson Refshauge, Michelle Watt, Margaret E. Mccully, Cheng HuangAbstract:A new method of sample preparation for cryo-scanning electron microscopy was used to visualize internal infection of wheat (Triticum aestivum) roots by the pathogenic fungus Rhizoctonia solani AG-8. The new method retained Fungal hyphae and root cells in situ in disintegrating root tissues, thus avoiding the distortions that can be introduced by conventional preparation by chemical fixation, dehydration and embedding. Infected roots frozen in liquid nitrogen were cryo-planed and etched (sublimed) at -80 degrees C for a critical length of time (up to 9 min) in the microscope column to reveal plant and Fungal Structures in three dimensions. Root and Fungal Structures were well preserved irrespective of infection severity. Root and hyphal cell walls were clearly seen and hyphal architecture within and between root cells was preserved. This rapid method permits three-dimensional in situ visualization of Fungal invasion within roots and has broad application for examination of diseases caused by other necrotrophic fungi.
Yves Piche - One of the best experts on this subject based on the ideXlab platform.
-
In vivo observations of the arbuscular mycorrhizal fungus Glomus mosseae in roots by confocal laser scanning microscopy
Fungal Biology, 1999Co-Authors: Horst Vierheiling, Christine Juge, Annette Böckenhoff, Michael Knoblauch, Florian M W Grundler, Yves Piche, Urs WyssAbstract:Confocal laser scanning microscopy was used for in vivo observations of autofluorescent Fungal Structures of Glomus mosseae. in intact, living rye-grass roots. Clear images of hyphae and collapsed arbuscules could be obtained without staining or sectioning roots. As rye-grass roots are highly transparent, with only a few cell layers above the vascular cylinder, they provide an excellent in vivo system for microscopical confocal laser scanning studies of arbuscular mycorrhizal Structures.
-
ink and vinegar a simple staining technique for arbuscular mycorrhizal fungi
Applied and Environmental Microbiology, 1998Co-Authors: Horst Vierheilig, Urs Wyss, Andrew P Coughlan, Yves PicheAbstract:We developed a reliable, inexpensive, and simple method for staining arbuscular-mycorrhizal Fungal colonizations in root tissues. Apart from applications in research, this nontoxic, high-quality staining method also could be of great utility in teaching exercises. After adequate clearing with KOH, an ink-vinegar solution successfully stained all Fungal Structures, rendering them clearly visible.
-
A modified procedure for staining arbuscular mycorrhizal fungi in roots
Zeitschrift für Pflanzenernährung und Bodenkunde, 1998Co-Authors: Horst Vierheilig, Yves PicheAbstract:Roots were cleared by boiling in 10% KOH and rinsed several times with tap water. Thereafter roots were boiled in the staining solutions. The staining solutions consisted in different dyes (trypban blue (0.05%), anilin blue (0.05%) and acid fuchsin (0.01%)) which were dissolved in usual household vinegar (5% acetic acid). For best contrast, tryphan and anilin blue stained roots were destained with tap water, whereas acid fuchsin stained roots were destained with vinegar. All Fungal Structures were stained and clearly visible.
P. Kolesik - One of the best experts on this subject based on the ideXlab platform.
-
Visualisation of mycorrhizal Fungal Structures and quantification of their surface area and volume using laser scanning confocal microscopy
Mycorrhiza, 1999Co-Authors: S. Dickson, P. KolesikAbstract:A method has been developed for the visualisation and three-dimensional (3D) measurement of mycorrhizal Fungal Structures inside plant roots. Sections of Allium porrum L. roots colonised by Glomus sp. 'City Beach' (WUM 16) and Lilium sp. roots colonised by Scutellospora calospora (Nicol. & Gerd.) Walker & Sanders (WUM 12(2)) were stained with acid fuchsin. This allowed fluorescence from the Fungal Structures to be observed under a laser scanning confocal microscope (LSCM) without interference from the plant cells. A series of horizontal optical sections were collected from a Glomus sp. arbuscule and from a hyphal coil of S. calospora. These data were used to produce extended focus images. Axial distortion in microscopic visualisation due to the refractive index mismatch between the immersion and mounting media was quantified using vertical scanning of the hyphae. A correction factor of 0.71 μm was used for the z-interval between the xy-slices. A series of binary xy-images from each structure was rendered into a 3D graphical model for viewing. The volume and surface area of the Structures were estimated using computerised 3D measurement and also by stereological integration of binary xy-images. With both Structures, the surface area estimates varied greatly between the two measuring systems, whereas differences in volume estimates were small. Computerised 3D measurement was considered more accurate than stereological integration of confocal binary images.
