Schematic representation of intravital microscopy protocols. Protocol 1 was designed to assess the action of MnBuOE and MnE on ongoing acute vaso-occlusive crises triggered by inflammation in TS mice. Anesthetized TS mice with window chamber implants were injected subcutaneously (SC) with 0.1, 0.2, or 2 mg/kg MnBuOE, 0.5 mg/kg or 2 mg/kg MnE, or vehicle (saline) 120 minutes after TNF-α challenge (time 0). Fifteen minutes before and 15 minutes after drug injection, blood cell behavior in the subdermal vasculature was recorded between the time points of 105 and 120 minutes (T₁₀₅ → T₁₂₀) and 135 and 225 minutes (T₁₃₅ → T₂₂₅), respectively. Protocol 2 was designed to assess the action of MnE on preventing precipitation of vaso-occlusion when given simultaneously with TNF-α, the inflammatory trigger of vaso-occlusion in TS mice. Anesthetized TS mice with window chamber implants were injected first with dyes for in vivo fluorescence cell labeling as described in “Materials and methods.” Thirty minutes after cell labeling, mice were SC administered 1 dose of 0.5 mg/kg MnE, or vehicle (saline), then challenged immediately with TNF-α (time 0). After 120 minutes, intravital microscopy was performed, and blood cell behavior in the subdermal vasculature was recorded between the time points of 120 and 180 minutes (T₁₂₀ → T₁₈₀). i.p., intraperitoneally; mAb, monoclonal antibody; PE, phycoerythrin.