Cells were pelleted and washed with 1× PBS solution for preparation of microscope slides. A thin 2% (w/v) agarose pad was prepared on a glass slide and 1 μL of cell suspension was added on top and air dried for about 5 min before placing a glass cover slip (Ke et al., 2016 (link)). Imaging was performed with a DMi8 automated inverted microscope (Leica Microsystems, #11889113) equipped with a CCD camera (Leica Microsystems, #DFC300 G), and a TXR (Leica Microsystems, #11525310) and YFP (Leica Microsystems, #11525306) filter cube. The Z-stack of captured images was processed in a blind deconvolution to remove out of focus fluorescence with the LAS X software version 3.3.3.16958 (Leica Microsystems, #11640612) with 3D deconvolution package (Leica Microsystems, #11640865).
Dmi8 automated inverted microscope
Manufactured by Leica
The DMi8 is an automated inverted microscope designed for advanced imaging applications. It features high-quality optics, motorized components, and integrated control software to enable precise and efficient sample examination.
Automatically generated - may contain errors
Lab products found in correlation
Las x software version 3, by Leica (2 mentions)
3d deconvolution package, by Leica (2 mentions)
Csu x1 m1n, by Yokogawa (2 mentions)
Ff01 650 13 25, by IDEX Corporation (2 mentions)
Ile 400, by Oxford Instruments (2 mentions)
Metamorph software, by Molecular Devices (2 mentions)
Hcx pl apo, by Leica (1 mentions)
5 protocols using dmi8 automated inverted microscope
1
Microscopy Slide Preparation and Imaging
2
Microscopic Analysis of Microbial Cultures
3
Microscopic Imaging of Bacterial Cells
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Cells were pelleted and washed with 1× PBS solution for preparation of microscope slides.
A thin 2 % (w/v) agarose pad was prepared on a glass slide and 1 µL of cell suspension was added on top and air dried for about 5 min before placing a glass cover slip (Ke et al., 2016) (link).Imaging was performed with a DMi8 automated inverted microscope (Leica Microsystems, #11889113) equipped with a CCD camera (Leica Microsystems, #DFC300 G), and a TXR (Leica Microsystems, #11525310) and YFP (Leica Microsystems, #11525306) filter cube. The Z-stack of captured images was processed in a blind deconvolution to remove out of focus fluorescence with the LAS X software version 3.3.3.16958 (Leica Microsystems, #11640612) with 3D deconvolution package (Leica Microsystems, #11640865).
A thin 2 % (w/v) agarose pad was prepared on a glass slide and 1 µL of cell suspension was added on top and air dried for about 5 min before placing a glass cover slip (Ke et al., 2016) (link).Imaging was performed with a DMi8 automated inverted microscope (Leica Microsystems, #11889113) equipped with a CCD camera (Leica Microsystems, #DFC300 G), and a TXR (Leica Microsystems, #11525310) and YFP (Leica Microsystems, #11525306) filter cube. The Z-stack of captured images was processed in a blind deconvolution to remove out of focus fluorescence with the LAS X software version 3.3.3.16958 (Leica Microsystems, #11640612) with 3D deconvolution package (Leica Microsystems, #11640865).
4
Live Chromatin Imaging with SiR-Hoechst
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Cells were placed in a 37 °C humid incubator by controlling the temperature and CO2 flow using H201-couple with temperature and CO2 units. Live chromatin imaging was performed using a DMI8 inverted automated microscope (Leica Microsystems) featuring a confocal spinning disk unit (CSU-X1-M1N, Yokogawa). An integrated laser engine (ILE 400, Andor) was used for excitation with a selected wavelength of 647 nm and 140 mW as excitation power. A 100× oil immersion objective (Leica HCX-PL-APO) with a 1.4 NA was chosen for a high-resolution imaging. Fluorescence emission of the SiR-Hoechst was filtered by a single-band bandpass filter (FF01-650/13-25, Semrock, Inc.). Image series of 150 frames (5 fps), with exposure time of 150 ms per frame, were acquired using Metamorph software (Molecular Devices) and detected using sCMOS cameras (ORCA-Flash4.0 V2) and 1 × 1 binning, with sample pixel size of 65 nm. All series were recorded at 37 °C.
5
Automated Confocal Imaging of DNA Dynamics
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DNA images were acquired using a DMI8 inverted automated microscope (Leica Microsystems) featuring a confocal spinning disk unit (CSU-X1-M1N, Yokogawa). Integrated laser engine (ILE 400, Andor) with a selected wavelength of 647 nm (140 mW) was used for excitation. Samples were imaged with an oil immersion objective (Leica HCX-PL-APO 100x/1.4 NA). Fluorescence emission of the SiR–Hoechst was filtered by a single-band bandpass filter (FF01-650/13-25, Semrock, Inc.). Image series of 150 frames (5 fps) were acquired using Metamorph software (Molecular Devices), and detected using sCMOS cameras (ORCA-Flash4.0 V2) and (1 × 1 binning), with sample pixel size of 65 nm. All series were recorded at 37°C and in a humid chamber by controlling the temperature and CO2 control flow using H201—couple with temperature and CO2 units.
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