Phagocytosis was assayed via uptake of pHrodo S. aureus BioParticles (Invitrogen, Eugene, OR). BioParticle conjugates are novel, no wash fluorogenic particles that are used for quantitative measurement of phagocytosis. Cells were cultured in complete media for 24 hours at a concentration of 2 × 105 in Mattek tissue culture dishes. BioParticles prepared per manufacturer instructions were added to cells at a 5:1 ratio and cells plus particles incubated at 37°C for 30 minutes to allow for uptake. Cells were washed to remove unattached BioParticles, fixed in 2% paraformalin, stained with Alexa-488 conjugated anti-MR antibody, and prepared for confocal microscopy. Samples were examined by confocal microscopy using anti-MR Alexa-488, or Alexa 647-conjugated antibody against anti-TLR2 or Rab antibodies.
Tissue culture dishes
Manufactured by MatTek
Tissue culture dishes are circular containers made of plastic or glass, designed for the in vitro cultivation of cells, tissues, or small organisms. They provide a sterile and controlled environment for the growth and maintenance of these biological samples.
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2 protocols using tissue culture dishes
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Quantitative Phagocytosis Assay Using pHrodo S. aureus
2
Quantitative Phagocytosis Assay Using pHrodo S. aureus
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Phagocytosis was assayed via uptake of pHrodo S. aureus BioParticles (Invitrogen, Eugene, OR). BioParticle conjugates are novel, no wash fluorogenic particles that are used for quantitative measurement of phagocytosis. Cells were cultured in complete media for 24 hours at a concentration of 2 × 105 in Mattek tissue culture dishes. BioParticles prepared per manufacturer instructions were added to cells at a 5:1 ratio and cells plus particles incubated at 37°C for 30 minutes to allow for uptake. Cells were washed to remove unattached BioParticles, fixed in 2% paraformalin, stained with Alexa-488 conjugated anti-MR antibody, and prepared for confocal microscopy. Samples were examined by confocal microscopy using anti-MR Alexa-488, or Alexa 647-conjugated antibody against anti-TLR2 or Rab antibodies.
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