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. Author manuscript; available in PMC: 2015 Aug 7.
Published in final edited form as: Nat Med. 2012 Sep;18(9):1386–1393. doi: 10.1038/nm.2847

Figure 6.

Figure 6

Immunofluorescence imaging of NETosis during human abscess formation. (a) Fresh abscess aspirates were stained with a PE-conjugated CD66b specific antibody, a PerCP-conjugated anti-CD45 specific antibody and SYTOX Green. The white arrow highlights an anuclear PMN adjacent to a NET that is outlined by a dotted line. The yellow arrow highlights a normal PMN with multilobar condensed nuclei. (b) Fresh live abscess aspirates stained with PE-conjugated CD16 specific antibody were directly injected into mouse skin to mimic our in vivo experiments and nuclei were pre-labeled with SYTO 60. NETs were visualized and quantified using the NET ratio. Exogenous DNase decreased visible NETs and decreased the NET ratio. (c) Normal PMN were stained with PE-conjugated CD16 specific antibody and SYTO 60. PMN were injected into mouse skin alone or with GFP-S. aureus. A NET is being released from the PMN stimulated with bacteria and the GFP-staphylococcus can be seen attached to the NET. NET release by in vivo human PMN is quantified as NET ratio (** P < 0.01 for untreated versus treated). (d) PMN nuclei were quantified in vivo from uninfected normal human PMN, human PMN infected with S. aureus and in human abscess PMN.