In this attempt to discover something "about the account upon which Respiration is so necessary to the Animals, that Nature hath furnish'd with Lungs", Boyle conducted numerous trials during which he placed a large variety of different creatures, including birds, mice, eels, snails and flies, in the vessel of the pump and studied their reactions as the air was removed.
Pada percobaan untuk menemukan "tentang hal penting bahwa Pernafasan sangat diperlukan pada Hewan, bahwa Alam telah memberikan Paru-paru", Boyle melakukan banyak uji coba yang mana ia menaruh berbagai macam makhluk hidup termasuk burung, tikus, belut, siput dan lalat, di dalam wadah pompa udara dan mempelajari reaksinya ketika udara dikeluarkan.