https://bridgeslab.sph.umich.edu/protocols/index.php?action=history&feed=atom&title=Measuring_Phagocytosis_using_pHrodo_Bioparticles 2026-06-02T02:42:56Z Revision history for this page on the wiki MediaWiki 1.45.1 https://bridgeslab.sph.umich.edu/protocols/index.php?title=Measuring_Phagocytosis_using_pHrodo_Bioparticles&diff=511&oldid=prev 2010-09-01T18:47:08Z

<p>wrote initial protocol</p> <p><b>New page</b></p><div>==Materials==<br /> *pHrodo BioParticles Conjugates (Invitrogen cat #A10010)<br /> *HBSS<br /> *OptiMEM<br /> <br /> ==Protocol==<br /> #Subculture cells 3-4 days in advance<br /> #On the day of the assay, harvest cells and spin down. Resuspend in OptiMEM at 1 000 000 cells/mL<br /> #Plate into a 96 well plate at 100 000 cells per well (100uL of a 1 000 000 cells/mL suspension), plating each condition in triplicate. Leave at least set of wells empty as a blank.<br /> #Add 100 uL of OptiMEM to blank wells.<br /> #Incubate at least 1h at 37C in a CO2 incubator.<br /> #Treat wells as desired.<br /> #Thaw one vial of particles for every 20 wells (including no cell controls). Add 2 mL HBSS to a vial and vortex briefly.<br /> #Transfer to a clean glass tube and sonicate for 5 min to ensure even dispersion.<br /> #After cells have adhered, aspirate media.<br /> #Replace media with 100 uL of the resuspended bioparticles.<br /> #Put in incubator for 2-3h<br /> #Scan plate using 550nm excitation and 600nm emission</div>

Davebrid