A Scalable Microfluidic Platform for Determining Cellular Heterogeneity by Copy Number Detection

A Scalable Microfluidic Platform for Determining Cellular Heterogeneity by Copy Number Detection



hello I'm Andrew price from technics genomics and I'm here today to talk about scaleable solution we have for calling copper number of variations on individual cells this product uses the 10x platform and so we start with individual cells or nuclei either from cell culture or isolated from individual tissues they entrapped in a microfluidic platform into a gel matrix which dates into what we call as a cell bead the cell beads allows us to process each cell individually to unpack it from its chromatin structure each cell is then partitioned into a second partition which is includes a 10x gel bead which allows individual barcoding of each cells that cell has then coalesced into a single phase which allows us to complete our 10x library which is sequence on to a standard Illumina platforms which allows us to call copy number variations with a turnkey pipeline and visualization software we can start with a variety of input cells including fresh tissue frozen tissue fresh tissue allows us to either use dissociated cells or allow us to isolate nuclei from them our frozen tissue you can get nuclei solution you can also use cultured cells or primary cells in this system it has a very low doublet rate which is essentially following poised on loading and so 1,000 cells generally has a double rate of under 2% the chemistry allows us very even profiling which has allows us to call individual copy number variations with minimal wasting in our sequencing our platform allows copy number or variation calling in on individual cells generally at the depth that we recommend sequencing – you can call it to achieve a resolution of around 2 mega bases on the individual cell however with clustering we were able to achieve individual resolution of 100 kilobases per cell here we show two references either cosmic or one that we generated an in-house from running pc our free true seek libraries and using the gingko platform we're using a gastric cancer cell line m'kay and 45 all the individual copy number variations to go into a real sample we have a tumor which we macro dissected into five different pieces overall we achieved over ten thousand individual cells that we sequenced of around 50 to 200 mega bases per cell looking at the entire tumor we were able to identify over five thousand cells which had copy number aberrations in it and further looking into that we were able to identify nine individual subclones that contain over eighty cells per clone as we can see look at each individual sections one through five is that each section varied not only in the amount of cancer cells that were detected from nine percent in one section to over eighty two percent at a different point of section but what is became clear is that if you look at the heterogeneity in the in the clonal structure of it that the the amounts of each clones varied tremendously from each of the individual cells so for example if we look at clone a we can see in one section it represented a very small part of the cancer cells that we put that over there we can see as we proceed through the tumor that a clone grew and to become a very large section at sections 3 and 4 over in section 5 again that became a smaller clone if we look at the other clones a through I we can see that they vary as we go from each different section and so for cold copy number variations the platform allows us to achieve at the recommended death to megabase clone an individual cell however in clustering cells together we able to achieve calling on a resolution down to 100 Kb and finally the the platform allows us to interrogate real samples as we're showing here in this case a breast cancer tumor and allow us to measure heterogeneity that exists within the tumor on a very easy and scalable solution for identifying thousands of cells at one time thank you for your attention and for if you have any further questions or want to learn more about this product please visit a 10x genomics website thank you

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