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Budapest, Hungary

Kiszler G.,University of Pecs | Kiszler G.,3DHISTECH Ltd. | Varhalmi E.,University of Pecs | Krecsak L.,Eotvos Lorand University | And 3 more authors.
Invertebrate Survival Journal | Year: 2016

The presence of γ-aminobutyric acid (GABA) immunoreactive elements was observed in the sensory system of Eisenia fetida (Annelida, Clitellata). Among the primary sensory cells a high number of labelled cells was found in the epithelium. Using whole-mount preparations and multispectral recording, the number and the distribution pattern of the immunopositive cells were determined in different body segments. Our morphological analysis revealed four typical types of stained primary sensory cells, which could be responsible for the different role of the GABA mediated sensory functions. The peripheral processes of the primary sensory cells ramifying at the border of epithelium and muscular layer produced a basiepidermal network where GABA-positive fibres and their connections were observed. The central processes of the labelled cells projected directly to the ventral nerve cord (VNC) through the segmental nerves to form the ventrolateral and ventromedial sensory longitudinal axon bundles (SLABs). Inhibitory GABA sensory inputs could influence indirectly the activity of the giant motoneurons through the ventrolateral giant axons, and thus the contraction of the body organizing withdrawal and escape reflexes. Applying ultrastructural investigations the synaptic connections of GABA-immunoreactive structures were identified both in the basiepidermal plexus, in the segmental nerves, and in the SLABs of VNC suggesting multistep regulatory effect of GABA in sensory processing of earthworms. © 2016, Universita degli Studi di Modena e Reggio Emilia. All Rights reserved. Source


Hegedus M.,3DHISTECH Ltd. | Varga V.S.,3DHISTECH Ltd. | Molnar B.,Hungarian Academy of Sciences
IFMBE Proceedings | Year: 2011

High-resolution scanning of the slides can be optimized based on tissue localization using low-resolution overview images in digital microscopy. Automatic preview image processing and tissue mapping can fail due to lack of sufficient image contrast in case of samples without visible dyes, e.g. for fluorescent samples or for samples prior to staining. We investigated the use of alternative imaging/illumination solutions (e.g. phase contrast, Schlieren, dark field). After examination of the Mie light scattering properties of tissue samples we developed and tested a method for the dark field preview illumination of glass slides based on our results. This solution proved to be well suited even for hydrated tissue samples prior to coverslipping and staining of any kind. Optimization of the staining procedure is also possible based on these overview images of the sample. Nevertheless, in the case of archived and contaminated slides tissue, dust and fingerprints were imaged with equal intensities. Therefore we proposed an off-axis thin line dark field illumination. This approach makes the separation of the images from different planes along the optical axis possible. Thus light scattered on the bottom of the glass slide, on the sample plane and on the surface of the coverslip is projected onto different pixels of the camera sensor and can be discriminated. We designed a compact light source and control program for testing this design and showed how the image of the tissue sample can be reconstructed from a series of pictures with minimizing the effect of contamination artifacts. © 2011 Springer-Verlag Berlin Heidelberg. Source


Micsik T.,Semmelweis University | Kiszler G.,3DHISTECH Ltd. | Szabo D.,3DHISTECH Ltd. | Krecsak L.,Podmaniczky u. 63 | And 4 more authors.
Pathology and Oncology Research | Year: 2015

HER2-positive breast cancers usually benefit from anti-HER2 therapy, thus, HER2 evaluation became inevitable for patient selection. HER2-negative (IHC 0, 1+) and strong positive (IHC 3+) cases can easily be interpreted with immunohistochemistry, but equivocal (IHC 2+) cases require further analysis of HER2 gene amplification using in situ hybridization. Our study aimed to validate digital pathology and automated image analysis for unbiased evaluation of HER2 immunostains. We developed an image segmentation algorithm for analyzing HER2-immunostaining (4B5 clone) in tissue microarrays of breast cancers. Two pathologists assessed 309 microscopic regions of at least 100 tumor cells each—representing all HER2 positivity groups—according to international guidelines either semi-quantitatively or by using the MembraneQuant software. Scoring results were statistically correlated with each other and with FISH data, and almost perfect agreement was found (inter-method Cohen’s kappa = 0.872, Spearman-rho = 0.928). When clinical relevance (scoring disagreement that may define erroneous treatment selection) was examined high agreement was found (quadratic weighted kappa = 0.967). Image analysis classified cases with excellent correlation with visual evaluation, therefore, MembraneQuant software proved to be a reliable tool for assessing HER2 immunoreactions and supporting better targeting anti-HER2 therapy. As digital analysis of immunomorphological markers allows permanent archiving, standardization and accurate reviewing of results, it supports quality assurance initiatives in diagnostic pathology—especially of equivocal cases which are hard to interpret. © 2015, Arányi Lajos Foundation. Source

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