RAW3mt cells were plated overnight on coverslips prior to imaging. Coverslips were washed 1x in Ringer Solution (155 mM NaCl, 4.5 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 5 mM HEPES, 10 mM D-glucose) and placed in the imaging chamber. Coverslips were imaged at 37°C ± 1°C. Microscopy was performed with an open pinhole on the Leica SP5 microscope with excitation from ‘white light’ and 488 nm argon lasers using Leica Applicate Suite Software (Leica). The images were processed using ImageJ software. Texas Red-conjugated zymosan A S. cerevisiae Bioparticles (ThermoFisher; Z2843) were fed to macrophages on coverslips and imaged for phagocytosis. “Whole cell mCa2+” was analyzed by drawing ROIs around individual cells during zymosan phagocytosis. For measurements of PPMiCa response, data analysis was performed as described in Extended Figure 5B .
Texas red conjugated zymosan a s cerevisiae bioparticles
Manufactured by Thermo Fisher Scientific
Texas Red-conjugated zymosan A S. cerevisiae Bioparticles are fluorescently labeled particles derived from the cell wall of the yeast Saccharomyces cerevisiae. The particles are conjugated with the Texas Red fluorescent dye, allowing for their detection and visualization using appropriate fluorescence microscopy or flow cytometry techniques.
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2 protocols using texas red conjugated zymosan a s cerevisiae bioparticles
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Calcium Imaging of Zymosan Phagocytosis
2
Calcium Imaging of Zymosan Phagocytosis
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RAW3mt cells were plated overnight on coverslips prior to imaging. Coverslips were washed 1x in Ringer Solution (155 mM NaCl, 4.5 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 5 mM HEPES, 10 mM D-glucose) and placed in the imaging chamber. Coverslips were imaged at 37°C ± 1°C. Microscopy was performed with an open pinhole on the Leica SP5 microscope with excitation from ‘white light’ and 488 nm argon lasers using Leica Applicate Suite Software (Leica). The images were processed using ImageJ software. Texas Red-conjugated zymosan A S. cerevisiae Bioparticles (ThermoFisher; Z2843) were fed to macrophages on coverslips and imaged for phagocytosis. “Whole cell mCa2+” was analyzed by drawing ROIs around individual cells during zymosan phagocytosis. For measurements of PPMiCa response, data analysis was performed as described in Extended Figure 5B .
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