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Dmi6000 inverted fluorescent microscope

Manufactured by Leica
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Sourced in Germany

The Leica DMI6000 is an inverted fluorescent microscope designed for advanced biological and biomedical research. It features a high-resolution optics system, motorized components, and integrated digital imaging capabilities for precise and efficient observation and analysis of fluorescent samples.

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Lab products found in correlation

Dfc365 fx camera, by Leica (1 mentions) Las x software, by Leica (1 mentions) Orca er b w ccd camera, by Hamamatsu Photonics (1 mentions) Dfc9000 scmos camera, by Leica (1 mentions) Dmi8 inverted fluorescent microscope, by Leica (1 mentions) Metamorph software, by Molecular Devices (1 mentions) Anti mouse igg h l alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 anti rabbit antibody, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

DMI6000 B inverted fluorescent microscope DMI6000 inverted microscope DMI6000 fluorescent microscope Inverted fluorescent microscope DMI6000 microscope Inverted microscope Fluorescent microscope DMI6000

4 protocols using dmi6000 inverted fluorescent microscope

1

Quantitative Fluorescence Microscopy Protocol

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Images were acquired with a Leica DMI 6000 inverted fluorescent microscope equipped with a 40× air lens and a Leica DFC365 FX camera (6.45 μm pixel size) using AF6000 Leica software v.3.1.0 (Leica Microsystems, Wetzlar, Germany). Twelve μm z-stacks (0.44 μm step size, 28 planes) were collected using the Cy5, Cy3, GFP, and DAPI channels (center/band width, nm: excitation 545/39, 620/60, 470/40, 360/40, respectively; emission 605/75, 700/75, 525/45, 470/40, respectively). Stacked images were presented as the maximum-intensity projection of the center five planes. All images were acquired with identical settings within each experiment. Cells that displayed oversaturated signal were excluded from analyses. For co-transfected conditions, only cells that were GFP positive and Cy3 positive were analyzed. The experimentalist was blinded to transfection conditions during the acquisition and data analyses for all the experiments are described below.
Kumar M.S., Fowler-Magaw M.E., Kulick D., Boopathy S., Gadd D.H., Rotunno M., Douthwright C., Golebiowski D., Yusuf I., Xu Z., Brown RH J.r., Sena-Esteves M., O’Neil A.L, & Bosco D.A. (2022). Anti-SOD1 Nanobodies That Stabilize Misfolded SOD1 Proteins Also Promote Neurite Outgrowth in Mutant SOD1 Human Neurons. International Journal of Molecular Sciences, 23(24), 16013.
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2

Fluorescence Microscopy Imaging Protocol

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Images were acquired on a DMI6000 inverted fluorescent microscope (Leica) using an ORCA-ER B/W CCD camera (Hamamatsu; native resolution 1344 × 1024 pixels) through Metamorph software (Molecular Devices, version 7.6) or on a DMI8 inverted fluorescent microscope (Leica; native resolution 2048 × 2048 pixels) using a Leica DFC9000 sCMOS camera through LASX software (Leica). Axons and soma were imaged using ×10 and ×20 air lenses. Images of axons showing CM-H2DCFDA, cyt c, and TMRE were imaged using ×63 oil lenses.
Geden M.J., Romero S.E, & Deshmukh M. (2021). p53 is required for nuclear but not mitochondrial DNA damage-induced degeneration. Cell Death & Disease, 12(1), 104.
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3

Stress Granule Formation in S2R+ Cells

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S2R+ cells were plated in 24-well plates in serum-free media on poly-l-lysine coated coverslips, and were treated with 1 ug dsRNA for 50 minutes, after which complete medium was added. Four days later, cells were stressed by replacing the media with fresh media containing 0.2 mM sodium arsensite (Sigma-Aldrich, St. Louis, MO). Two hours after the initiation of the sodium arsenite treatment, cells were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS). The rest of the immunofluorescence protocol was identical to that with the HEK cells. Primary antibodies used: anti-dFMR1 (5A11, 1:200, DSHB). Secondary antibodies used: anti-Mouse IgG (H+L) Alexa Fluor 594 (#A-11005, 1:1000, Invitrogen, Carlsbad, CA). Images were acquired with Leica DMI6000 inverted fluorescent microscope with an oil immersion X100 objective and 1.6 magnification factor. Quantification of stress granules was performed with CellProfiler.
Berson A., Sartoris A., Nativio R., Van Deerlin V., Toledo J.B., Porta S., Liu S., Chung C.Y., Garcia B.A., Lee V.M., Trojanowski J.Q., Johnson F.B., Berger S.L, & Bonini N.M. (2017). TDP-43 promotes neurodegeneration by impairing chromatin remodeling. Current biology : CB, 27(23), 3579-3590.e6.
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4

Imaging and Quantifying Neutrophil Extracellular Traps

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Cells or NETs were fixed with 2% paraformaldehyde solution and labeled with granular protein and nuclear dye. Myeloperoxidase (MPO), a primary granule protein, was labeled using primary antibody against MPO overnight at 4 degrees followed by secondary Alexa Fluor 488 anti-rabbit antibody for 1 h at room temperature (Both obtained from Invitrogen, Waltham, MA, USA). Further nuclei were stained using DAPI containing mounting media. Only secondary antibody was used as specificity control. The co-localization of MPO and DNA was used for the identification of NETs. Images were captured and analyzed at 40× objective using 2 filters- DAPI, FITC on a Leica DMI6000 inverted fluorescent microscope.
Singhal A., Yadav S., Chandra T., Mulay S.R., Gaikwad A.N, & Kumar S. (2022). An Imaging and Computational Algorithm for Efficient Identification and Quantification of Neutrophil Extracellular Traps. Cells, 11(2), 191.
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