Dmi6000b widefield microscope

Manufactured by Leica

Sourced in United Kingdom, Germany

The DMI6000B is a widefield microscope manufactured by Leica. It is designed for general laboratory applications and provides a range of imaging capabilities. The microscope features a modular design and can be configured with various objective lenses and accessories to suit different research needs.

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Lab products found in correlation

Las x, by Leica (7 mentions) Volocity 6, by Quorum Technologies (3 mentions) Tcs sp8 confocal laser scanning microscope, by Leica (3 mentions) Perfection v850 pro flatbed scanner, by Leica (2 mentions) Rabbit anti gfp, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Goat serum, by Vector Laboratories (1 mentions) Mouse anti pan cytokeratin, by Abcam (1 mentions) Tcs sp8 laser, by Leica (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Ti2 inverted microscope, by Nikon (1 mentions) Ckx53 inverted, by Olympus (1 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions) Ckx53, by Olympus (1 mentions) Sp8 confocal microscope, by Leica (1 mentions) Hoechst 33258, by Merck Group (1 mentions) Tcs spe confocal microscope, by Leica (1 mentions) Secondary fluorophore conjugated antibodies, by Jackson ImmunoResearch (1 mentions) Triton x 100, by Merck Group (1 mentions) Las x software, by Leica (1 mentions)

Spelling variants of this product

DMI6000B (1065 citations) DMI6000B microscope (503 citations) Widefield microscope (14 citations) DMI6000B widefield epifluorescence microscope (2 citations) DMI6000B inverted widefield fluorescence microscope (2 citations)

18 protocols using dmi6000b widefield microscope

Immunofluorescence Analysis of Humanized Mouse Thymus

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Half of the thymic graft from each humanized mouse was fixed in 4% paraformaldehyde for 30 min at room temperature, then transferred to 30% glucose PBS at 4°C overnight. Tissues were then embedded in OCT compound and frozen. Cryosections (5 μm) were blocked with goat serum (Vector Laboratories) for 15 min, and stained with following primary antibodies: rabbit anti-GFP (Millipore/Sigma) and mouse anti-Pan-cytokeratin (Abcam) in room temperature for 3 h or 4°C overnight. Sections were subsequently stained with AF488 rabbit anti-GFP IgG (H+L) antibody (Jackson ImmunoResearch) and AF647 goat anti-mouse IgG (H+L) antibody (Jackson ImmunoResearch) at room temperature for 1 h. All sections were mounted in mounting medium with DAPI (Vector Laboratories). Images were obtained using Leica DMI 6000B wide field microscope.

Li Y., Teteloshvili N., Tan S., Rao S., Han A., Yang Y.G, & Creusot R.J. (2019). Humanized Mice Reveal New Insights Into the Thymic Selection of Human Autoreactive CD8+ T Cells. Frontiers in Immunology, 10, 63.

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Microscopic Imaging for Graft Analysis

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Images were captured using either a flatbed scanner Epson Perfection V850 PRO, a Leica DMI6000B widefield microscope or a Leica TCS SP8 laser scanning confocal microscope. The image acquisition software was Leica LAS X and images were processed using Volocity 6.5.1 (Quorum Technologies) and Adobe Photoshop. Any adjustments were applied equally across the entire image, and without the loss of any information. The following images were digitally stitched from multiple images: Figs. 1b, 2h, 3a–f, 4b, c, Supplementary Figs. 1k, 5b and 7d, e, g–j. Figures 1a and 4a were created by the authors. Each image represents a typical result obtained from analysis of minimum 2–3 graft sections per animal.

Tiklová K., Nolbrant S., Fiorenzano A., Björklund Å.K., Sharma Y., Heuer A., Gillberg L., Hoban D.B., Cardoso T., Adler A.F., Birtele M., Lundén-Miguel H., Volakakis N., Kirkeby A., Perlmann T, & Parmar M. (2020). Single cell transcriptomics identifies stem cell-derived graft composition in a model of Parkinson’s disease. Nature Communications, 11, 2434.

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Endothelial Cell Migration Assay

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GECs were seeded (1 × 10⁴ cells, each side) into Ibidi Culture-Inserts (Ibidi). and cultured until 95% confluence was reached. After that, the inserts were removed, and cells were stained with 1 mg/mL Calcein AM (Thermo Fisher Scientific) for 30 min at 37 °C. Then, fresh EndoPM was added in the presence of pharmacological treatments, as previously described. After 24 h, images of GECs that migrated into the cells-free gap were acquired with an inverted Leica DMI6000B widefield microscope at 20× magnifications in five random fields. Cells migrated into the gap were than counted using “Analyze Particles” in ImageJ.

Guarnaccia L., Navone S.E., Masseroli M.M., Balsamo M., Caroli M., Valtorta S., Moresco R.M., Campanella R., Schisano L., Fiore G., Galiano V., Garzia E., Appiani G.C., Locatelli M., Riboni L, & Marfia G. (2022). Effects of Metformin as Add-On Therapy against Glioblastoma: An Old Medicine for Novel Oncology Therapeutics. Cancers, 14(6), 1412.

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Immunostaining of Thymic Graft Sections

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hES-cultures in 24-well tissue culture plates were fixed with paraformaldehyde in PBS (4%) for 10 minutes at room temperature. Cells were washed in PBS twice, permeabilized in PBS with 0.1% triton for 20 min, and blocked in 5% fetal donkey serum for 1 hour at room temperature.
Thymic grafts were extracted, embedded in OCT (Tissue-Tec, Torrance CA) media, frozen and 5–7um thick sections cut for immune staining. Sections were stained with H&E to visualize gross histology and interface of the thymic graft with the mouse renal tissue. For immunofluorescent staining, tissue sections were fixed and permeabilized in 100% ice-cold acetone and allowed to dry completely. Tissue sections were blocked in PBS supplemented with 0.1% Tween and 0.1% Bovine Serum Albumin. Slides were washed in PBS 0.1% Tween and stained with primary antibody for 2 hours at room temperature, and then washed and incubated in secondary antibodies for 2 hours at room temperature.
Cultures or tissue sections were incubated with one, or a combination of two or three of the following primary antibodies and appropriate secondary antibodies listed in Supplemental Table II. Images were collected on a Leica SCN 400 whole slide scanning platform for H&E stained sections and immunofluorescent images were collected on a Leica TCS SP8 2 photon laser scanning or Leica DMI 6000B wide field microscope.

Gras-Pena R., Danzl N.M., Khosravi-Maharlooei M., Campbell S.R., Ruiz A.E., Parks C.A., Savage W.M., Holzl M.A., Chatterjee D, & Sykes M. (2021). Human Stem Cell-Derived Thymic Epithelial Cells Enhance Human T Cell Development in a Xenogeneic Thymus. The Journal of allergy and clinical immunology, 149(5), 1755-1771.

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Imaging Protocols for Microscopy Research

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Images were captured using either an Epson Perfection V850 PRO flatbed scanner, a Leica DMI6000B widefield microscope, or a Leica TCS SP8 confocal laser scanning microscope. Image acquisition software was Leica LAS X and images were processed using Volocity 6.5.1 software (Quorum Technologies) and Adobe Photoshop. Any adjustments were applied equally across the entire image and without the loss of any information.

Fiorenzano A., Birtele M., Wahlestedt J.N, & Parmar M. (2021). Evaluation of TH-Cre knock-in cell lines for detection and specific targeting of stem cell-derived dopaminergic neurons. Heliyon, 7(1), e06006.

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Imaging Protocols for Microscopy Analysis

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Images were captured using an Epson Perfection V850 PRO flatbed scanner, a Leica DMI6000B widefield microscope, or a Leica TCS SP8 confocal laser-scanning microscope, or a Nikon inverted Ti2 microscope equipped with a CSU-W1 spinning-disk system. Image acquisition software was Leica LAS X and images were processed using Volocity 6.5.1 software (Quorum Technologies) and Adobe Photoshop. Any adjustments were applied equally across the entire image, and without the loss of any information.

Fiorenzano A., Sozzi E., Birtele M., Kajtez J., Giacomoni J., Nilsson F., Bruzelius A., Sharma Y., Zhang Y., Mattsson B., Emnéus J., Ottosson D.R., Storm P, & Parmar M. (2021). Single-cell transcriptomics captures features of human midbrain development and dopamine neuron diversity in brain organoids. Nature Communications, 12, 7302.

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Brightfield and Fluorescent Microscopy Protocol

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Brightfield images were taken using an Olympus CKX53 inverted microscope with a 4x objective (UPlanFL N 4x/0.13 NA) and acquired with the OLYMPUS cell Sens Standard v2.3 software. Fluorescent images were captured within a week from staining using a Leica DMI6000B widefield microscope or a Leica TCS SP8 laser scanning confocal microscope. The image acquisition software was Leica LAS X and images were processed using Adobe Photoshop CC 2020. In comparison experiments, all the images were taken with the same software settings and any adjustments were applied equally across the entire image, without the loss of any information.

Sozzi E., Kajtez J., Bruzelius A., Wesseler M.F., Nilsson F., Birtele M., Larsen N.B., Ottosson D.R., Storm P., Parmar M, & Fiorenzano A. (2022). Silk scaffolding drives self-assembly of functional and mature human brain organoids. Frontiers in Cell and Developmental Biology, 10, 1023279.

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Fluorescence Imaging of Arl3-GFP Subcellular Localization

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Cells in exponential growth phase (OD₆₀₀ 0.8–1.2) were pelleted and washed three times in 1x PBS, 2% glucose. Fluorescence images were acquired in the washing solution on a Leica DMI6000 B widefield microscope as previously described39 (link). Recorded images were processed using Photoshop CS5 (Adobe Systems, San Jose, CA, USA). Arl3-GFP subcellular localization was characterized as normal (punctate) or mislocalization (other). More than 500 cells were counted per strain.

Osberg C., Aksnes H., Ninzima S., Marie M, & Arnesen T. (2016). Microscopy-based Saccharomyces cerevisiae complementation model reveals functional conservation and redundancy of N-terminal acetyltransferases. Scientific Reports, 6, 31627.

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Immunofluorescence Assay for Phosphorylated p65

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HUVECs grown on coverslips etched with hydrofluoric acid were fixed with 4% PFA (Electron Microscopy Science) in phosphate-buffered saline (PBS; 20 min, 20°C), washed once in PBS (5 min, 20°C), permeabilized using 0.5% Triton X-100 in PBS (5 min, 20°C) and blocked with 1% bovine serum albumin (BSA) in PBS (Sigma-Aldrich; 45 min, 20°C). Phosphorylated (at Ser536) p65 was detected using a rabbit monoclonal antibody (1:500 dilution, 0.5% BSA in PBS; #04-1000, Millipore, Nottingham, UK) and Alexa488-conjugated donkey anti-rabbit AffinityPure F(ab’)2 Fragment (1.5 μg/ml; Jackson ImmunoResearch, Maine, USA). After DAPI counter-staining, images were collected on a Leica DMI6000 B widefield microscope and analysed using ImageJ [60 ]; nuclei were encircled, the mean intensity calculated per area, and nuclear fluorescence (arbitrary units) calculated by subtracting the background (measured as the minimum intensity in the image).

Diermeier S., Kolovos P., Heizinger L., Schwartz U., Georgomanolis T., Zirkel A., Wedemann G., Grosveld F., Knoch T.A., Merkl R., Cook P.R., Längst G, & Papantonis A. (2014). TNFα signalling primes chromatin for NF-κB binding and induces rapid and widespread nucleosome repositioning. Genome Biology, 15(12), 536.

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Fluorescent Imaging and Confocal Microscopy

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Fluorescent images were captured using a Leica DMI6000B wide-field microscope. Image acquisition software was Leica LAS X and images were processed using Adobe Photoshop CC 2018. Any adjustments were applied equally across the entire image and without the loss of any information. Immunohistochemical stainings were analyzed using a Leica confocal microscope with 20×/0.70 IMM and HP PL APO CS2 63×/1.20 water objectives. Double staining was confirmed by conduction of high-magnified confocal z-stacks. All figures were assembled using Canvas software.

Birtele M., Storm P., Sharma Y., Kajtez J., Wahlestedt J.N., Sozzi E., Nilsson F., Stott S., He X.L., Mattsson B., Ottosson D.R., Barker R.A., Fiorenzano A, & Parmar M. (2022). Single-cell transcriptional and functional analysis of dopaminergic neurons in organoid-like cultures derived from human fetal midbrain. Development (Cambridge, England), 149(23), dev200504.

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