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Delgado, Ryan

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Delgado

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Delgado, Ryan
Author's Publications: search.filters.isAuthorOfPublication.362ecfea-a874-41f8-aff9-8e525d810678

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    Thermal-plex: fluidic-free, rapid sequential multiplexed imaging with DNA-encoded thermal channels
    (Springer Science and Business Media LLC, 2023-12-27) Hong, Fan; Kishi, Jocelyn; Delgado, Ryan; Jeong, Jiyoun; Saka Kirli, Sinem; Su, Hanquan; Cepko, Constance; Yin, Peng
    Multiplexed fluorescence imaging is typically limited to three- to five-plex on standard setups. Sequential imaging methods based on iterative labeling and imaging enable practical higher multiplexing, but generally require a complex fluidic setup with several rounds of slow buffer exchange (tens of minutes to an hour for each exchange step). We report the thermal-plex method, which removes complex and slow buffer exchange steps and provides fluidic-free, rapid sequential imaging. Thermal-plex uses simple DNA probes that are engineered to fluoresce sequentially when, and only when, activated with transient exposure to heating spikes at designated temperatures (thermal channels). Channel switching is fast (<30 s) and is achieved with a commercially available and affordable on-scope heating device. We demonstrate 15-plex RNA imaging (five thermal × three fluorescence channels) in fixed cells and retina tissues in less than 4 min, without using buffer exchange or fluidics. Thermal-plex introduces a new labeling method for efficient sequential multiplexed imaging.
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    Single-cell delineation of lineage and genetic identity in the mouse brain
    (Springer Science and Business Media LLC, 2021-12-15) Bandler, Rachel; Vitali, Ilaria; Delgado, Ryan; Ho, May; Dvoretskova, Elena; Ibarra Molinas, Josue; Frazel, Paul; Mohammadkhani, Maesoumeh; Machold, Robert; Maedler, Sophia; Liddelow, Shane; Nowakowski, Tomasz; Fishell, Gordon; Mayer, Christian
    During neurogenesis, mitotic progenitor cells lining the ventricles of the embryonic mouse brain undergo their final rounds of cell division, giving rise to a wide spectrum of postmitotic neurons and glia. The link between developmental lineage and cell type diversity remains an open question. Here we used massively parallel tagging of progenitors to track clonal relationships and transcriptomic signatures during murine forebrain development. We quantified clonal divergence and convergence across all major cell classes postnatally, and found diverse types of GABAergic neurons that share a common lineage. Divergence of GABAergic clones occurred during embryogenesis upon cell-cycle exit, suggesting that differentiation into subtypes is initiated as a lineage-dependent processes at the progenitor cell level.