Macrophages were plated onto 8-chambered coverglass (LabTek, Thermoscientific, Rochester, NY) (13 (link), 22 (link), 23 (link)). Cells were incubated with stimuli at 37°C for specified times. Coverglass were then mounted on Nikon Ti-E inverted microscope equipped with CSU-X1 confocal spinning-disk head (Yokogawa, Sugarland, TX). A coherent, 4 Watt laser (Coherent, Santa Clara, CA) was used as an excitation light source to produce excitation wavelengths 488, 568, and 647 nm using an acoustic optical tunable tuner. To acquire high-quality fluorescence images, a high-magnification, high-numerical aperture objective was used (Nikon, 1003, 1.49 numerical aperture, oil immersion) was used. A polarizer (Nikon, MEN 51941) and Wollaston prisms (Nikon, MBH76190) were used to acquire differential interference contrast (DIC) images. Emission light from the sample was collected after passage through the appropriate emission filters (Semrock, Rochester, NY). Images were acquired using an EM-CCD camera (Hamamatsu, C9100-13, Bridgewater, NJ). Image acquisition was performed using MetaMorph software (Molecular Devices, Dowington, PA). Raw image data files were processed using Adobe Photoshop CS4 and assembled in Adobe Illustrator, version CS4 (Adobe Systems, San Jose, CA).
Mbh76190
Manufactured by Nikon
The MBH76190 is a piece of laboratory equipment manufactured by Nikon. It is designed to perform a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach. Additional information about the intended use or interpretation of the product's features is not available.
Automatically generated - may contain errors
Lab products found in correlation
Men 51941, by Nikon (2 mentions)
Lab tek, by Thermo Fisher Scientific (2 mentions)
Metamorph software, by Molecular Devices (2 mentions)
C9100 13, by Hamamatsu Photonics (1 mentions)
Ti e inverted microscope, by Yokogawa (1 mentions)
Csu x1 spinning disk confocal head, by Yokogawa (1 mentions)
Em ccd camera, by Hamamatsu Photonics (1 mentions)
2 protocols using mbh76190
1
High-Resolution Live-Cell Imaging of Macrophages
2
Live-cell Imaging of C. glabrata Phagocytosis
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2 × 104 peritoneal macrophages were plated onto eight-chambered coverslip slides (LabTek, Thermo Scientific, Rochester, NY, USA) in RPMI-complete. CFSE-labeled C. glabrata were added at MOI 1:1, spun at 700 g × 1 min for immediate co-culture and slides mounted on a Nikon Ti-E inverted microscope equipped with an EM-CCD camera (Hamamatsu Photonics K.K., Hamamatsu, Japan). The excitation source was an 89 North MultiLine LaserBank (89 North, Burlington, VT, USA) A piezo stage (Prior Instruments, Rockland, MA, USA) capable of X, Y, and Z movement was used for acquisition. A polarizer (Nikon, MEN 51941) and Wollaston prisms (Nikon, MBH76190) were used to acquire differential interference contrast (DIC) images. Emission light from the samples was collected after passage through the appropriate emission filters (Semrock, Rochester, NY, USA) (32 (link)). Images were acquired using MetaMorph software (Molecular Devices, Downingtown, PA, USA), and processed using Adobe Photoshop CS5 and assembled in Adobe Illustrator, version CS5 (Adobe Systems, San Jose, CA, USA).
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