Dmi8 microscope

Manufactured by Yokogawa

The DMi8 is a high-performance microscope designed for laboratory use. It features a modular design and a range of available accessories to support various microscopy techniques. The core function of the DMi8 is to provide clear, detailed imaging for scientific observation and analysis.

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Lab products found in correlation

Ixon ultra 897, by Oxford Instruments (3 mentions) Borealis, by Oxford Instruments (3 mentions) Hcx pl apo 1.4 0 70na oil objective lens, by Leica (3 mentions) Csu w1, by Yokogawa (3 mentions) Nucblue liveprobe reagent, by Thermo Fisher Scientific (2 mentions) Metamorph software v7, by Molecular Devices (2 mentions) Orca flash 4.0 camera, by Hamamatsu Photonics (2 mentions) Metamorph, by Molecular Devices (1 mentions)

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DMi8 inverted microscope (5 citations)

7 protocols using dmi8 microscope

Live Imaging of C. elegans Oocytes

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All imaging was carried out using a Leica DMi8 microscope outfitted with a spinning disk confocal unit–CSU-W1 (Yokogawa) with Borealis (Andor), dual iXon Ultra 897 (Andor) cameras, and a 100x HCX PL APO 1.4–0.70NA oil objective lens (Leica). Metamorph (Molecular Devices) imaging software was used for controlling image acquisition. The 488nm and 561nm channels were imaged simultaneously every 10 seconds with 1μm Z-spacing (either 16μm or 21μm total Z-stacks depending on the fluorescent markers used, with the same stack size used for all movies utilizing the same fluorescent markers).
In utero live imaging of oocytes was accomplished by mounting adult worms with a single row or less of embryos in 1.5μl of M9 mixed with 1.5μl of 0.1μm polystyrene Microspheres (Polysciences Inc.) on a 6% agarose pad with a coverslip gently laid over top. Ex utero imaging of oocytes was carried out by cutting open adult worms with a single row or less of embryos in 4μl of egg buffer (118mM NaCl, 48mM KCl, 2mM CaCl₂, 2mM MgCl₂, and 0.025 mM of HEPES, filter sterilized before HEPES addition) on a coverslip before mounting onto a 2% agarose pad on a microscope slide.

Schlientz A.J, & Bowerman B. (2020). C. elegans CLASP/CLS-2 negatively regulates membrane ingression throughout the oocyte cortex and is required for polar body extrusion. PLoS Genetics, 16(10), e1008751.

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Visualizing Lysosomal Tubule Formation

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The formation of tubules in late endosomes/lysosomes was followed by live imaging of cells expressing LAMP1-mcherry at 37 °C and 5% CO2 using a Leica DMi8 microscope equipped with a Yokogawa Confocal Spinning Disk module. Cells were chosen randomly, with the only criterion being LAMP1-mCherry levels sufficiently high to detect lysosomal tubules.

Boutry M., Pierga A., Matusiak R., Branchu J., Houllegatte M., Ibrahim Y., Balse E., El Hachimi K.H., Brice A., Stevanin G, & Darios F. (2019). Loss of spatacsin impairs cholesterol trafficking and calcium homeostasis. Communications Biology, 2, 380.

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Chromatin Flow Dynamics Imaging

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Cells were plated in Fluorodishes (Worldprecision; Cat#: FD35100) at the rate of 2 × 10⁵ cells per dish the day before performing the live imaging. Thirty minutes before live imaging, the medium was replaced with 1 ml of fresh medium with two drops of NucBlue LiveProbe reagent (Thermo Fisher; Cat#: R37605). Live imaging was performed on a Leica DMI8 microscope, equipped with a CSU-X1 Yokogawa spinning disk module. The acquisition was realized with a 40× dry objective (N.A. 1.40) and collected by the Hamamatsu Orca flash 4.0 camera. The microscope was controlled by the Metamorph software v7.10.2.240 from Molecular Devices. Images were acquired in brightfield (one plane, exposure time 50 ms) with a 405 nm laser at 15% power (Z stack, seven planes centered on the middle plane of the nuclei, Z step of 0.5 μm, exposure time 150 ms) every 2 min for 1.5–2 h. Analysis was performed on the best Z plane chosen for each stage position, using a set of custom-written Fiji scripts. Briefly, the nuclei were segmented and registered for XY translation and rotation using the MultiStackReg plugin. The chromatin flow was then measured using the PIV plugin on the first 10 time points, restricted to the nuclei. The PIV magnitude value was obtained by averaging the values measured for the entire nucleus over the 10 time points.

Zakharova V.V., Magnitov M.D., Del Maestro L., Ulianov S.V., Glentis A., Uyanik B., Williart A., Karpukhina A., Demidov O., Joliot V., Vassetzky Y.S., Mège R.M., Piel M., Razin S.V, & Ait-Si-Ali S. (2022). SETDB1 fuels the lung cancer phenotype by modulating epigenome, 3D genome organization and chromatin mechanical properties. Nucleic Acids Research, 50(8), 4389-4413.

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Confocal Imaging of Cellular Structures

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Confocal images were acquired on a Leica DMi8 microscope equipped with Yokogawa CSU-W1 Spinning Disk Confocal and Andor Zyla 4.2 sCMOS camera controlled with Andor Fusion software. Images were acquired in 0.1 μm sections (for deconvolution) or 0.2 μm otherwise with a 100× objective (NA 1.4). When indicated, images were deconvolved in Fusion software on default settings. For co-localization, signals from TetraSpeck beads included in the mounting media were used as positive control.

Smith C.E., Tsai Y.C., Liang Y.H., Khago D., Mariano J., Li J., Tarasov S.G., Gergel E., Tsai B., Villaneuva M., Clapp M.E., Magidson V., Chari R., Byrd R.A., Ji X, & Weissman A.M. (2021). A structurally conserved site in AUP1 binds the E2 enzyme UBE2G2 and is essential for ER-associated degradation. PLoS Biology, 19(12), e3001474.

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Quantitative Live Imaging of Caenorhabditis elegans

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All imaging was carried out using a Leica DMi8 microscope outfitted with a spinning disk confocal unit–CSU-W1 (Yokogawa) with Borealis (Andor), dual iXon Ultra 897 (Andor) cameras, and a 100x HCX PL APO 1.4–0.70NA oil objective lens (Leica). (Molecular Devices) imaging software was used for controlling image acquisition. The 488nm and 561nm channels were imaged simultaneously with 1um Z-spacing.
Ex utero live imaging used the following parameters: 1μm Z spacing, 16 focal planes, 100ms exposure, 10s interval. Imaging was carried out by dissecting worms in 3ul egg salt buffer (118mM NaCl, 48mM KCl, 2mM CaCl2, 2mM MgCl2, and 0.025 mM of HEPES, filter sterilized before HEPES addition) on a coverslip before mounting onto a 2% agarose pad (diluted in egg salt buffer) on a microscope slide. Upon imaging for AID knockdown strains, worms were dissected onto egg salt buffer with 1 mM auxin and mounted onto a 2% agarose pad (diluted in egg salt buffer) with 1 mM auxin.
In utero live imaging used the following parameters: 1 μm Z spacing, 20 μm stacks, 80s exposure, 20s interval. Imaging was carried out by placing adult worms in 1.5 μl of M9 mixed with 1.5 μl of 1 μm polysterene microspheres (Polysciences Inc.) on a coverslip which was mounted carefully onto a 5% agarose pad.

Quiogue A.R., Sumiyoshi E., Fries A., Chuang C.H, & Bowerman B. (2023). Microtubules oppose cortical actomyosin-driven membrane ingression during C. elegans meiosis I polar body extrusion. PLOS Genetics, 19(10), e1010984.

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Imaging Caenorhabditis elegans Embryogenesis

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All imaging was carried out using a Leica DMi8 microscope outfitted with a spinning disk confocal unit–CSU-W1 (Yokogawa) with Borealis (Andor), dual iXon Ultra 897 (Andor) cameras, and a 100x HCX PL APO 1.4–0.70NA oil objective lens (Leica). (Molecular Devices) imaging software was used for controlling image acquisition. The 488nm and 561nm channels were imaged simultaneously with 1um Z-spacing.
Ex utero live imaging used the following parameters: 1μm Z spacing, 16 focal planes, 100ms exposure, 10s interval. Imaging was carried out by dissecting worms in 3ul egg salt buffer (118mM NaCl, 48mM KCl, 2mM CaCl2, 2mM MgCl2, and 0.025 mM of HEPES, filter sterilized before HEPES addition) on a coverslip before mounting onto a 2% agarose pad (diluted in egg salt buffer) on a microscope slide.
In utero live imaging used the following parameters: 1 μm Z spacing, 20 μm stacks, 80s exposure, 20s interval. Imaging was carried out by placing adult worms in 1.5 μl of M9 mixed with 1.5 μl of 1 μm polysterene microspheres (Polysciences Inc.) on a coverslip which was mounted carefully onto a 5% agarose pad.

Quiogue A.R., Sumiyoshi E., Fries A., Chuang C.H, & Bowerman B. (2023). Cortical microtubules oppose actomyosin-driven membrane ingression during C. elegans meiosis I polar body extrusion. bioRxiv.

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Live Imaging of Chromatin Dynamics

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Cells were plated in Fluorodishes (Worldprecision; Cat#: FD35100) at the rate of 2 x 10 5 cells per dish the day before performing the live imaging. 30 min before live imaging, the medium was replaced with 1 mL of fresh medium with 2 drops of NucBlue LiveProbe reagent (Thermo Fisher; Cat#: R37605). Live imaging was performed on a Leica DMI8 microscope, equipped with a CSU-X1 Yokogawa spinning disk module. The acquisition was realized with a 40x dry objective (N.A. 1.40) and collected by the Hamamatsu Orca flash 4.0 camera. The microscope was controlled by the Metamorph software v7.10.2.240 from Molecular Devices. Images were acquired in brightfield (one plane, exposure time 50 ms) with a 405 nm laser at 15 % power (Z stack, 7 planes centered on the middle plane of the nuclei, Z step of 0.5 µm, exposure time 150 ms) every 2 min for 1.5 to 2 hours. Analysis was performed on the best Z plane chosen for each stage position, using a set of custom-written Fiji scripts. Briefly, the nuclei were segmented and registered for XY translation and rotation using the MultiStackReg plugin. The chromatin flow was then measured using the PIV plugin on the first 10 time points, restricted to the nuclei. The PIV magnitude value was obtained by averaging the values measured for the entire nucleus over the 10 time points.

Zakharova V., Magnitov M., Del-Maestro L., Ulianov S.V., Glentis A., Ulyanik B., Williart A., Karpukhina A., Demidov O.N., Joliot V., Vassetzky Y., Mège R., Piel M., Razin S.V., & Ait‐Si‐Ali S. (2021). SETDB1 Fuels the Lung Cancer Phenotype by Modulating Epigenome, 3D Genome Organization and Chromatin Mechanical Properties.

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