microscopy techniques Dotaz Zobrazit nápovědu

Kreft (Ljubjana) -- 14.00 -14.20 Rescuing real time three-dimensional scanning electron microscopy (L7 Boyde (London) -- 14.20 - 14.40 Review of recent trends in optical micromanipulation techniques (L8) Brno) -- 15.20-15.30 Visualisation of kidney glomeruli in real time video-rate confocal reflection microscopy

Cytokinematics 2006 Hradec Králové, Česko
Autor Autorita
Univerzita Karlova. Lékařská fakulta v Hradci Králové. Ústav lékařské biologie a genetiky
Autor Autorita

Hradec Králové : Department of Medical Biology and Genetics LF UK, 2006

42 s. ; 30 cm

Konspekt
Buněčná biologie. Cytologie
NLK Obory
biologie
cytologie, klinická cytologie
radiologie, nukleární medicína a zobrazovací metody
histologie

Článek

Comparing Super-Resolution Microscopy Techniques to Analyze Chromosomes

The importance of fluorescence light microscopy for understanding cellular and sub-cellular structures ...

International journal of molecular sciences. 2021 ; 22 (4) : . [pub] 20210214

Int J Mol Sci
ISSN 1422-0067
Medvik
Zdroj

The importance of fluorescence light microscopy for understanding cellular and sub-cellular structures and functions is undeniable. However, the resolution is limited by light diffraction (~200-250 nm laterally, ~500-700 nm axially). Meanwhile, super-resolution microscopy, such as structured illumination microscopy (SIM), is being applied more and more to overcome this restriction. Instead, super-resolution by stimulated emission depletion (STED) microscopy achieving a resolution of ~50 nm laterally and ~130 nm axially has not yet frequently been applied in plant cell research due to the required specific sample preparation and stable dye staining. Single-molecule localization microscopy (SMLM) including photoactivated localization microscopy (PALM) has not yet been widely used, although this nanoscopic technique allows even the detection of single molecules. In this study, we compared protein imaging within metaphase chromosomes of barley via conventional wide-field and confocal microscopy, and the sub-diffraction methods SIM, STED, and SMLM. The chromosomes were labeled by DAPI (4',6-diamidino-2-phenylindol), a DNA-specific dye, and with antibodies against topoisomerase IIα (Topo II), a protein important for correct chromatin condensation. Compared to the diffraction-limited methods, the combination of the three different super-resolution imaging techniques delivered tremendous additional insights into the plant chromosome architecture through the achieved increased resolution.

MeSH
chromozomy rostlin chemie genetika metabolismus MeSH
DNA-topoisomerasy typu II metabolismus MeSH
fluorescenční barviva chemie MeSH
fluorescenční mikroskopie metody MeSH
indoly chemie MeSH
ječmen (rod) cytologie genetika MeSH
konfokální mikroskopie metody MeSH
metafáze genetika MeSH
reprodukovatelnost výsledků MeSH
zobrazení jednotlivé molekuly metody MeSH
Publikační typ
časopisecké články MeSH
srovnávací studie MeSH

Článek

Identification of pollen taxa by different microscopy techniques

alternative method for visual inspection is automated pollen analysis based on the image analysis technique ...

PLoS One. 2021 ; 16 (9) : e0256808. [pub] 20210901

ISSN 1932-6203
Medvik
Zdroj

Melissopalynology is an important analytical method to identify botanical origin of honey. Pollen grain recognition is commonly performed by visual inspection by a trained person. An alternative method for visual inspection is automated pollen analysis based on the image analysis technique. Image analysis transfers visual information to mathematical descriptions. In this work, the suitability of three microscopic techniques for automatic analysis of pollen grains was studied. 2D and 3D morphological characteristics, textural and colour features, and extended depth of focus characteristics were used for the pollen discrimination. In this study, 7 botanical taxa and a total of 2482 pollen grains were evaluated. The highest correct classification rate of 93.05% was achieved using the phase contrast microscopy, followed by the dark field microscopy reaching 91.02%, and finally by the light field microscopy reaching 88.88%. The most significant discriminant characteristics were morphological (2D and 3D) and colour characteristics. Our results confirm the potential of using automatic pollen analysis to discriminate pollen taxa in honey. This work provides the basis for further research where the taxa dataset will be increased, and new descriptors will be studied.