The Experts below are selected from a list of 2376 Experts worldwide ranked by ideXlab platform
Maxim Rubin-blum - One of the best experts on this subject based on the ideXlab platform.
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Fluorescence in situ hybridization (FISH) images of methanotrophic (MOX, MITC849 probe double-labelled with 594 dye) symbionts in 10 μm sections of H. (S.) methanophila sponge (Mictlan Knoll individual).
2018Co-Authors: Maxim Rubin-blumAbstract:Twenty 3-channel images were acquired with Zeiss Axioplan 2 Epifluorescence Microscope (Zeiss, Jena, Germany) with 10x magnification. Color legend is: MOX, magenta; DNA (DAPI), blue; autofluoresence at ~520 nm with ~490 nm excitation (FITC filter), green. These images were aligned with FIJI Grid/collection stitching plugin to produce the resulting mosaic.
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Additional FISH images of methanotrophic (MOX, MITC849 probe double-labelled with 594 dye) symbionts in 10 μm sections of H. (S.) methanophila sponge (Mictlan Knoll individual), Epifluorescence microscpopy
2018Co-Authors: Maxim Rubin-blumAbstract:We double-labelled MTC849 (5` – CGTTAGCTCCACCACTAAG – 3`) with Atto594 dye (Biomers, Ulm, Germany), and hybridized it with sponge tissue in 20% formamide buffer using standard protocol (Duperron et al., 2008). Photomicrographs were acquired with Zeiss Axioplan 2 Epifluorescence Microscope (Zeiss, Jena, Germany), 40x or 100x lenses (see file name). Three channels were used - 594 (MITC849), DAPI and FITC(autofluoresence)
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Fluorescence in situ hybridization (FISH) images of methanotrophic (MOX, MITC849 probe double-labelled with 594 dye) symbionts in 10 μm sections of I. methanophila sponge (Chapopote Knoll).
2018Co-Authors: Maxim Rubin-blumAbstract:We double-labelled MTC849 (5` – CGTTAGCTCCACCACTAAG – 3) with Atto594 dye (Biomers, Ulm, Germany), and hybridized it with sponge tissue in 20% formamide buffer using standard protocol (Duperron et al., 2008). Photomicrographs were acquired with confocal laser-scanning Microscope (LSM 780, Carl Zeiss, Germany) - named CLSM*** or with Zeiss Axioplan 2 Epifluorescence Microscope (Zeiss, Jena, Germany) - named 7-108-ROV17*** . Three channels were acquired - 594, DAPI and 488/FITC (autofluoresence). Five images that had been acquired with 10x magnification have been used to reconstruct the overview representation
Gregory R Monteith - One of the best experts on this subject based on the ideXlab platform.
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an automated Epifluorescence microscopy imaging assay for the identification of phospho akt level modulators in breast cancer cells
Journal of Pharmacological and Toxicological Methods, 2018Co-Authors: Elke Kaemmerer, Dane Turner, Amelia A Peters, Sarah J Robertsthomson, Gregory R MonteithAbstract:AKT is an enzyme of the PI3K/pAKT pathway, regulating proliferation and cell survival. High basal levels of active, phosphorylated AKT (pAKT) are associated with tumor progression and therapeutic resistance in some breast cancer subtypes, including HER2 positive breast cancers. Various stimuli can increase pAKT levels and elevated basal pAKT levels are a feature of PTEN-deficient breast cancer cell lines. The aim of this study was to develop an assay able to identify modulators of pAKT levels using an automated Epifluorescence Microscope and high content analysis. To develop this assay, we used HCC-1569, a PTEN-deficient, HER2-overexpressing breast cancer cell line with elevated basal pAKT levels. HCC-1569 cells were treated with a selective pharmacological inhibitor of AKT (MK-2206) to reduce basal pAKT levels or EGF to increase pAKT levels. Immunofluorescence images were acquired using an automated Epifluorescence Microscope and integrated intensity of cytoplasmic pAKT staining was calculated using high content analysis software. Mean and median integrated cytoplasmic intensity were normalized using fold change and standard score to assess assay quality and to identify most robust data analysis. The highest z' factor was achieved for median data normalization using the standard score method (z' = 0.45). Using our developed assay we identified the calcium homeostasis regulating proteins TPRV6, STIM1 and TRPC1 as modulators of pAKT levels in HCC-1569 cells. Calcium signaling controls a diverse array of cellular processes and some calcium homeostasis regulating proteins are involved in modulating pAKT levels in cancer cells. Thus, these identified hits present promising targets for further assessment.
Elke Kaemmerer - One of the best experts on this subject based on the ideXlab platform.
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an automated Epifluorescence microscopy imaging assay for the identification of phospho akt level modulators in breast cancer cells
Journal of Pharmacological and Toxicological Methods, 2018Co-Authors: Elke Kaemmerer, Dane Turner, Amelia A Peters, Sarah J Robertsthomson, Gregory R MonteithAbstract:AKT is an enzyme of the PI3K/pAKT pathway, regulating proliferation and cell survival. High basal levels of active, phosphorylated AKT (pAKT) are associated with tumor progression and therapeutic resistance in some breast cancer subtypes, including HER2 positive breast cancers. Various stimuli can increase pAKT levels and elevated basal pAKT levels are a feature of PTEN-deficient breast cancer cell lines. The aim of this study was to develop an assay able to identify modulators of pAKT levels using an automated Epifluorescence Microscope and high content analysis. To develop this assay, we used HCC-1569, a PTEN-deficient, HER2-overexpressing breast cancer cell line with elevated basal pAKT levels. HCC-1569 cells were treated with a selective pharmacological inhibitor of AKT (MK-2206) to reduce basal pAKT levels or EGF to increase pAKT levels. Immunofluorescence images were acquired using an automated Epifluorescence Microscope and integrated intensity of cytoplasmic pAKT staining was calculated using high content analysis software. Mean and median integrated cytoplasmic intensity were normalized using fold change and standard score to assess assay quality and to identify most robust data analysis. The highest z' factor was achieved for median data normalization using the standard score method (z' = 0.45). Using our developed assay we identified the calcium homeostasis regulating proteins TPRV6, STIM1 and TRPC1 as modulators of pAKT levels in HCC-1569 cells. Calcium signaling controls a diverse array of cellular processes and some calcium homeostasis regulating proteins are involved in modulating pAKT levels in cancer cells. Thus, these identified hits present promising targets for further assessment.
Gaku Mizuguchi - One of the best experts on this subject based on the ideXlab platform.
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fast multicolor 3d imaging using aberration corrected multifocus microscopy
Nature Methods, 2013Co-Authors: Sara Abrahamsson, Jiji Chen, Bassam Hajj, Sjoerd Stallinga, Alexander Y Katsov, Jan Wisniewski, Gaku MizuguchiAbstract:Conventional acquisition of three-dimensional (3D) microscopy data requires sequential z scanning and is often too slow to capture biological events. We report an aberration-corrected multifocus microscopy method capable of producing an instant focal stack of nine 2D images. Appended to an Epifluorescence Microscope, the multifocus system enables high-resolution 3D imaging in multiple colors with single-molecule sensitivity, at speeds limited by the camera readout time of a single image.
Sara Abrahamsson - One of the best experts on this subject based on the ideXlab platform.
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fast multicolor 3d imaging using aberration corrected multifocus microscopy
Nature Methods, 2013Co-Authors: Sara Abrahamsson, Jiji Chen, Bassam Hajj, Sjoerd Stallinga, Alexander Y Katsov, Jan Wisniewski, Gaku MizuguchiAbstract:Conventional acquisition of three-dimensional (3D) microscopy data requires sequential z scanning and is often too slow to capture biological events. We report an aberration-corrected multifocus microscopy method capable of producing an instant focal stack of nine 2D images. Appended to an Epifluorescence Microscope, the multifocus system enables high-resolution 3D imaging in multiple colors with single-molecule sensitivity, at speeds limited by the camera readout time of a single image.