50’000 AM were FACS sorted and resuspended in 50 uL PBS. Before starting, slides were cleaned from dust by dipping them into 96% ethanol and let dry on the bench. Slides were assembled into cytoclips together with the required tissue and cytofunnel. 50 uL of cell suspension were applied into the funnel and subsequently centrifuged for 5 minutes at 800g in a Sandon Cytospin 2 machine. After disassembling, slides were dried overnight at room temperature protected from dust and H&E staining was performed the next day with the RAL Diff-Quick Kit following manufacturer’s instructions.
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How to cite:
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
Piattini, F and Kopf, M(2022). Staining of cytospin preparations of AM populations. Bio-protocol Preprint. bio-protocol.org/prep1806.
Li, F., Piattini, F., Pohlmeier, L., Feng, Q., Rehrauer, H. and Kopf, M.(2022). Monocyte-derived alveolar macrophages autonomously determine severe outcome of respiratory viral infection. Science Immunology 7(73). DOI: 10.1126/sciimmunol.abj5761
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