Dmi 6000 cs inverted microscope

Manufactured by Leica camera

Sourced in United Kingdom, Germany

The Leica DMI 6000 CS is an inverted microscope designed for advanced cell and tissue culture research. The microscope features a modular design, allowing for customization to meet specific research needs. The core function of the DMI 6000 CS is to provide high-quality imaging and analysis capabilities for a variety of specimen types.

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Lab products found in correlation

Tcs sp5 confocal laser scanning microscope, by Leica Microsystems (3 mentions) Anti ipla2β, by Cayman Chemical (2 mentions) Af488 conjugated goat anti mouse igg, by Thermo Fisher Scientific (2 mentions) Af594 conjugated goat anti rabbit, by Thermo Fisher Scientific (2 mentions) μ slide vi0, by Ibidi (2 mentions) Tcs sp5 microscope, by Leica camera (2 mentions) Las af software, by Leica Microsystems (2 mentions) Fv1000, by Olympus (1 mentions) Af594, by Thermo Fisher Scientific (1 mentions) Mz16 stereomicroscope, by Leica camera (1 mentions) Sytox green, by Thermo Fisher Scientific (1 mentions) To pro 3 iodide, by Thermo Fisher Scientific (1 mentions) Rat tail collagen type 1, by BD (1 mentions) Labtek glass slides, by Thermo Fisher Scientific (1 mentions) Gapdh, by Merck Group (1 mentions)

8 protocols using dmi 6000 cs inverted microscope

Immunohistochemical Imaging of Focal Adhesions

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Glass coverslips were coated with human fibronectin (10 μg/ml) in Ca²⁺/Mg²⁺‐PBS. WM266.4 and 451Lu cells seeded at low confluency were fixed in PBS‐4% PFA‐4% sucrose for 10 min, quenched with PBS‐glycine 0.1 M for 30 min, then permeabilized and blocked with PBS‐10% FCS‐0.2% saponin for 15 min. Primary antibodies diluted in PBS‐5% FCS‐0.2% saponin incubation lasted 2 h. For immunodetection, secondary antibodies (diluted 1/1,000) were incubated for 30 min at 37°C. F‐actin was detected using Acti‐stain. Nuclei were stained with DAPI at 1 μg/ml for 15 min. Coverslips were mounted in Vectashield. Fluorescence micrographs were taken using laser scanning confocal systems (TCS SP8 model mounted on a DMI 6000 CS inverted microscope, Leica or IX81‐based Olympus FV1000). For FAs counting (vinculin), fixation was done in 70% ethanol and no saponin was used.

Shoji K.F., Bayet E., Leverrier‐Penna S., Le Devedec D., Mallavialle A., Marionneau‐Lambot S., Rambow F., Perret R., Joussaume A., Viel R., Fautrel A., Khammari A., Constantin B., Tartare‐Deckert S, & Penna A. (2023). The mechanosensitive TRPV2 calcium channel promotes human melanoma invasiveness and metastatic potential. EMBO Reports, 24(4), e55069.

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Visualizing Monocyte FPR2 and iPLA2β

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Primary human monocytes were plated in 6 channel micro-slides (μ-Slide VI 0.4, iBidi GmbH, Germany), with hrANXA1 being added to one reservoir only. Cells were fixed with 2% formaldehyde in 0.1M PBS prior to immunostaining with mouse monoclonal anti-FPR2 (Genovac, Freiburg, Germany) or rabbit polyclonal anti-iPLA₂β, (Cayman Chemical, Ann Arbor, MI), followed by secondary labelling with either AF488-conjugated goat anti-mouse IgG or AF594-conjugated goat anti-rabbit (both Invitrogen, UK). Cells were then further labelled with 5U/ml AF594 or AF488-conjugated phalloidin (Invitrogen, UK) counterstained with DAPI and examined using a TCS SP5 confocal laser scanning microscope (Leica Microsystems) fitted with 405nm, 488nm and 594nm lasers, and attached to a Leica DMI6000CS inverted microscope fitted with a 63× oil immersion objective lens (NA, 1.4mm; working distance, 0.17mm). Images were captured with Leica LAS AF 2.6.1 software and analyzed by using ImageJ software (National Institutes of Health).

McArthur S., Gobbetti T., Kusters D.H., Reutelingsperger C.P., Flower R.J, & Perretti M. (2015). Definition of a Novel Pathway Centered on Lysophosphatidic Acid To Recruit Monocytes during the Resolution Phase of Tissue Inflammation. The Journal of Immunology Author Choice, 195(3), 1139-1151.

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Immunofluorescence Microscopy of Cells

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Cells were fixed with 4% PFA and stained with the indicated antibodies. Fluorescence microphotographs were taken using a laser scanning confocal system (TCS SP8 model mounted on a DMI 6000 CS inverted microscope, Leica, Heidelberg, Germany). Images were taken using a 63x oil-immersion objective (1.4 N.A.), collected in a format of 1,024 ×1,024 pixels and analyzed using the ImageJ software (NIH, Bethesda, Maryland, USA).

Kenji S.F., Kurma K., Collet B., Oblet C., Debure L., Di Primo C., Minder L., Vérité F., Danger Y., Jean M., Penna A., Levoin N, & Legembre P. (2022). MMP7 cleavage of amino-terminal CD95 death receptor switches signaling toward non-apoptotic pathways. Cell Death & Disease, 13(10), 895.

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Nuclei Isolation and Confocal Imaging

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For confocal microscopy, nuclei were isolated from oocytes of 0.5–1.5 mm in diameter by jeweler forceps in the isolation medium “5:1” (described above) under the observation at Leica MZ16 stereomicroscope. Isolated nuclei were incubated for 5 min in “5:1” medium containing 0.07 μM Sytox Green (Molecular Probes) [34 (link)]. Confocal laser scanning microscopy was carried out with a Leica TCS SP5 microscope based on a Leica DMI 6000 CS inverted microscope. Specimens were examined by the XYZ scanning technique using HC PL APO 20× objective and argon laser (496 nm). Images were obtained using LAS AF software (Leica Microsystems, Germany), and 3D reconstruction was processed with Imaris 5.0.1 (Bitplane, AG) software.

Dedukh D., Litvinchuk S., Rosanov J., Mazepa G., Saifitdinova A., Shabanov D, & Krasikova A. (2015). Optional Endoreplication and Selective Elimination of Parental Genomes during Oogenesis in Diploid and Triploid Hybrid European Water Frogs. PLoS ONE, 10(4), e0123304.

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Quantifying Viral Spread in Hippocampus

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Viral spread was assessed from three brain sections (6 dentate gyri) per animal from a 1 in 18 coronal series through the hippocampus. tdTomato expression, indicative of cre recombinase activity, was used as a proxy marker for viral infection of the dentate gyrus. Expression was scored using a semiquantitative 0-4 scale ranging from 0 (0%), 1 (<25%), 2 (25-50%), 3 (50-75%), and 4 (>75%) of dentate granule cells expressing the tdTomato reporter. Scoring was conducted by an investigator blind to treatment group using a Leica DMI6000 CS inverted microscope under 10X magnification. Scores reflect estimates of the percentage of granule cells in the section regardless of whether labeling was dispersed through the upper and lower blades or concentrated in a portion of the dentate. Viral expression was also evident in CA1 pyramidal cells, CA3 pyramidal cells, cortical excitatory cells, and thalamic neurons. To characterize this expression, six brain sections per animals from the 1 in 18 coronal series were scored by an investigator blind to group using a Nikon AXR Inverted microscope under 10X magnification. Animals were scored as having expression in a region if one or more brain sections contained tdTomato labeled cells. Animal scores (expression present/not present) were compiled to generate the percentage of animals in each study group that had some expression in the four regions.

Godale C.M., Parkins E.V., Gross C, & Danzer S.C. (2022). Impact of Raptor and Rictor deletion on hippocampal pathology following status epilepticus. Journal of molecular neuroscience : MN, 72(6), 1243-1258.

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Imaging Monocyte FPR2 and iPLA2β

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Primary human monocytes were plated in six channel microslides (μ-Slide VI 0.4; iBidi), with hrANXA1 being added to one reservoir only. Cells were fixed with 2% formaldehyde in 0.1 M PBS prior to immunostaining with mouse monoclonal anti-FPR2 (Genovac, Freiburg, Germany) or rabbit polyclonal anti-iPLA₂β (Cayman Chemical, Ann Arbor, MI), followed by secondary labeling with either AF488-conjugated goat anti-mouse IgG or AF594-conjugated goat anti-rabbit (both Invitrogen). Cells were then further labeled with 5 U/ml AF594- or AF488-conjugated phalloidin (Invitrogen) counterstained with DAPI and examined using a TCS SP5 confocal laser-scanning microscope (Leica Microsystems) fitted with 405, 488, and 594 nm lasers, and attached to a Leica DMI6000CS inverted microscope fitted with a 63× oil immersion objective lens (NA, 1.4 mm; working distance, 0.17 mm). Images were captured with Leica LAS AF 2.6.1 software and analyzed by using ImageJ software.

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Oocyte Nucleus Imaging in Poeciliopsis

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In addition to the classical analysis of diplotene chromosomal spread, we investigated the intact oocyte nucleus from P. formosa and P. mexicana individuals. Nuclei were isolated with fine forceps from oocytes of 0.5–1 mm diameter in isolation medium “5:1”. Isolated nuclei were transferred to isolation medium containing 1 µM TO-PRO™-3 Iodide (Thermo Fisher Scientific). Confocal laser scanning microscopy was conducted with a Leica TCS SP5 microscope based on a Leica DMI 6000 CS inverted microscope. Specimens were examined by the XYZ scanning technique using HC PL APO 20 × objective and HeNe laser (633 nm).
Images were captured and processed using LAS AF software (Leica Microsystems, Germany); 3D reconstructions were made using Imaris 5.0.1 (Bitplane, AG) software.

Dedukh D., da Cruz I., Kneitz S., Marta A., Ormanns J., Tichopád T., Lu Y., Alsheimer M., Janko K, & Schartl M. (2022). Achiasmatic meiosis in the unisexual Amazon molly, Poecilia formosa. Chromosome Research, 30(4), 443-457.

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Annexin A1 Localization and Protein Analysis

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For ANXA1 analysis the membrane-bound fraction was extracted by EDTA-EGTA wash (1mM in PBS), as previously described (21) ; total protein extraction was performed by 3 cycles of freeze/thawing in RIPA buffer (1mM EDTA, 150mM NaCl, 50mM Tris⋅HCl, 1% Triton X-100, 0.1% SDS, 1% sodium deoxycholate, protease inhibitors, pH 7.4). Protein resolution and visualisation was performed as previously described (9, 20, 22) , using the following antibodies: Annexin A1 (1:5000; Invitrogen), ℗-S-Annexin A1 [1:1000; (22, 11) and GAPDH (1:5000; Sigma)]. Blots were analysed with NIH Image J 1.42 software. hCMEC/D3 cells were cultured on Lab-tek glass slides (Thermo Scientific, UK) pre-coated with rat tail type I collagen (BD Biosciences, San Jose CA, US), following published protocols (9, 23) and using primary antibodies directed against occludin (1:200; Invitrogen) and zonula occludens-1 (ZO-1; 1:100; Invitrogen). Nuclei were counterstained with DAPI.
Images were captured by using a TCS SP5 confocal laser scanning microscope (Leica Microsystems) fitted with 405-nm, 488-nm, 594nm and 633-nm lasers, and attached to a Leica DMI6000CS inverted microscope fitted with 63X oil immersion objective lens (NA, 1.4 mm, working distance, 0.17 mm). Images were captured with Leica LAS 2.6.1 software and analysed by using ImageJ software (National Institutes of Healths).

Maggioli E., McArthur S., McArthur S., Mauro C., Kieswich J., Kusters D.H.M., Reutelingsperger C.P.M., Yaqoob M, & Solito E. (2016). Estrogen protects the blood-brain barrier from inflammation-induced disruption and increased lymphocyte trafficking. Brain, behavior, and immunity, 51.

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