PCBC Multiplexed in situ Hybridization Workshop (Materials and Recordings)

With the support of the NHLBI Progenitor Cell Biology Consortium (PCBC), Mark Krasnow, Tushar Desai, Pehr Harbury, and colleagues hosted a “PCBC Multiplexed in situ Hybridization Workshop” on March 22, 2017, at Stanford University.

Workshop Blurb:

Single cell transcriptional profiling in situ is of high interest to biologists because it enables spatial localization of specific cell populations within intact tissues, allowing emergent identification of dynamic signaling interactions of these cells with their neighbors. For stem cell biologists, this approach can provide insights into how stem cells are regulated within their in vivo niches, and the ability to interrogate genes “on-demand,” without being limited to markers for which suitable antibodies are available, is particularly valuable. Proximity ligation- in situ hybridization (PLISH) is a next-generation, single cell resolution, multiplexed technology that is cheap, rapid, simple, and does not require any specialized equipment. It is biologist “friendly” and amenable to automated generation of single cell expression profiles displayed as a heat map, in which individual cells can be mapped back onto the intact tissue. Up to five transcripts can be simultaneously detected, and iterative rounds of staining, imaging, and erasing can be performed to increase the multiplexing. PLISH was invented in the PCBC and this one-day, hands-on training workshop will provide PCBC members with access to this powerful technology before its publication.

Multiplexed in situ hybridization using PLISH. Six channel in situ hybridization of embryonic day 18.5 mouse
lung marks distinct cell populations during alveolar differentiation. Br, bronchiole.

Workshop Materials and Recordings:

Workshop materials are attached and below is a link to the workshop recordings for your reference:

https://www.youtube.com/playlist?list=PL0QrRbpf8KbSL7xnKZz62rMNJlWxGQJLw

Workshop Forum:

Open forum for your comments/questions. You must have a PCBC user account to access this forum. Contact Andrea Lefever (alefever@som.umaryland.edu) or Ling Tang (litang@som.umaryland.edu) for a user account.

 

 

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