Mice were sensitized to chicken Ova from Sigma-Aldrich (Oakville, ON, Canada) using the protocol reported by Jacobsen et al.5 (link) with the modifications described here. Briefly, Ova was prepared at 0.909 mg/mL in PBS and mixed as per the manufacturer’s instructions with Imject Alum (Thermo Scientific, Waltham, MA, USA) at a ratio of 11:14 of Ova to Imject Alum. Mice were first sensitized to Ova by IP injection of 100 µL of either PBS or Ova/Imject Alum mix on days 0 and 14. On days 24, 25, and 26, mice were challenged with a 25-min exposure to nebulized 1 mg/mL Ova in saline. For the challenge, mice were housed in a pie cage (Braintree Scientific, Braintree, MA, USA) that received nebulized Ova from a Schuco cup nebulizer, with pressure on the vacuum pump set at <1 kg/cm2. Mice were imaged on day 28.
Acute silica exposure was performed similar to Ferreira et al.60 (link). Briefly, mice were anesthetized with isofluorane and then instilled intranasally with sterile saline alone or sterile saline containing 10 mg crystalline silica with a particle size of 0.5–10 μm (Sigma Chemical, St Louis, MO). Twelve hours later, animals were prepared for lung intravital microscopy as described above.
Acute silica exposure was performed similar to Ferreira et al.60 (link). Briefly, mice were anesthetized with isofluorane and then instilled intranasally with sterile saline alone or sterile saline containing 10 mg crystalline silica with a particle size of 0.5–10 μm (Sigma Chemical, St Louis, MO). Twelve hours later, animals were prepared for lung intravital microscopy as described above.
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