Dr. Peter Adams, Sanford Burnham Prebys Medical Discovery Institute
Dr. Geoffrey Wahl, Salk Institute for Biological Studies
Dr. Brian James, Sanford Burnham Prebys Medical Discovery Institute
Single Cell Sequencing Workshop Group/Social
Date: First Monday Monthly
Time: 4:00pm - 5:00pm
Sanford Burnham Prebys Medical Discovery Institute
Fishman Auditorium, Building #4
10901 North Torrey Pines Road
La Jolla, CA 92037
Monday, February 4, 2019 CANCELLED
Monday, March 4, 2019
Presenter: Wes Goldman, Technical Sales Specialist, Digital Spatial Profiling at nanoString Technologies
Title: Morphology Driven High-Plex Spatial Analysis of Tissue Microenvironments with NanoString’s Digital Spatial Profiling
Summary: Historically, immunohistochemistry and immunofluorescence have been used to assess spatial heterogeneity of proteins and nucleic acids in tissue slices. However, these techniques are of limited utility due to restricted dynamic range, difficult quantitation, and limited multiplexing capability.
NanoString’s GeoMx™ Digital Spatial Profiler* combines standard immunofluorescence techniques with digital optical barcoding technology to perform highly multiplexed, spatially resolved profiling experiments. In a single reaction, the DSP technology performs whole slide imaging with up to four fluorescent stains to capture tissue morphology and select regions of interest for high plex profiling of proteins and RNA.
Monday, April 1, 2019
Presenter: Calum Marrs, MSci, Regional Executive – West USA at Domomite & Dolomite Bio, a Blacktrace brand
Title: Nadia Introduction
Summary: Nadia takes scRNA-Seq protocols to the next level by using automation, flexibility and an open platform to generate high quality reproducible single cell data at low cost. Nadia is an automated, microfluidic droplet-based platform for single cell research that encapsulates up to 8 samples, in parallel, in under 20 mins. Over 50,000 single cells can be captured per cartridge in a run. Nadia guides users through all relevant steps of protocols via an easy-to-use touchscreen interface, integrating sample stirring and temperature control. Adding the Nadia Innovate upgrades to the Nadia instrument into the ultimate tool for the development of novel single-cell protocols and applications.
2018 Workshop Group/Social Dates
Monday, March 5, 2018
Presenter: Sebastian A. Preissl, Ph.D., Associate Director, Single Cell Genomics under Dr. Bing Ren at the UCSD Center for Epigenomics in the Department of Cellular and Molecular Medicine
Title: Single nucleus analysis of chromatin accessibility in mouse for brain development
Summary: The forebrain plays a key role in multiple higher-order brain functions including memory and consciousness. Transcriptional regulatory regions in the genome including promoters and distal acting enhancers play fundamental roles for forebrain development. Regulatory elements can be identified by the presence of open chromatin as measured by ATAC-seq (Assay for transposase-accessible chromatin using sequencing). However, unraveling the role of regulatory DNA elements has proven challenging for two main reasons: 1) regulatory DNA elements are dynamic and thus their activity must be studied across multiple stages; 2) tissues are heterogeneous and comprised of different cell types with cell-type-dependent chromatin accessibility patterns. To overcome these challenges we have optimized a combinatorial barcoding assisted assay for transposase accessible chromatin for frozen brain tissue samples. Analysis of more than 15,000 forebrain nuclei at eight developmental stages reveals 20 distinct cell populations corresponding to major neuronal and non-neuronal cell types. In addition, we define cell-type-specific transcriptional regulatory sequences, infer potential master transcriptional regulators and delineate developmental changes in forebrain cellular composition.
Monday, April 2, 2018 Cancelled
Monday, May 7, 2018 Cancelled
Presenter: Alain Domissy, Ph.D., Scientific Director, Center for Computational Biology and Bioinformatics at The Scripps Research Institute
Title: The Metadata is your Program
Summary: We will present a new way for experimental scientists to efficiently manage their experiments metadata and as a result gain the power to autonomously trigger and control both upstream compute intensive as well more interactive downstream analysis from the comfort of just their laptop's text editor, browser and email programs.
This will be illustrated with an example of a 10x single cell rna-seq experiment. We will use a tutorial dataset which is a downsized version of a published paper, and allows going through the complete pipeline experience in less then one hour.
Monday, June 4, 2018
Presenter Raj Giraddi, Ph.D., Research Associate, Gene Expression Laboratory, Salk Institute
Title: Single cell analysis of the stem cell states
Summary: We generated a single cell transcriptome data set encompassing fetal, postnatal and adult mouse mammary epithelium and paid special attention to the perinatal interval, over which the prevalent, multipotent fMaSC phenotype declines and differentiation ensues. We elucidate biological programs that distinguish bipotent stem cells from differentiating and lineage-committed cells. Our results reveal the surprising nature of the mammary stem cell states and changes how we view the mammary gland hierarchy and its subsequent implications in breast cancer.
Monday, July 2, 2018 CANCELLED
Monday, August 6, 2018 CANCELLED
Monday, September 3, 2018 CANCELLED
Monday, October 1, 2018
Presenter: Abhishek Sohni, postdoctoral fellow in Dr. Miles Wilkinson’s laboratory, University of California, San Diego (UCSD)
Title: Neonatal and Adult Human Testis Defined at the Single-Cell Level
Summary: Over 100 million men worldwide suffer from infertility. In most cases, the underlying cause is unknown. Treating human infertility requires a deep understanding of human spermatogenesis, but most of what we know about spermatogenesis comes from investigations in mice and other rodents. Given that human spermatogenesis significantly differs from rodent spermatogenesis in several aspects, we elected to perform a detailed analysis of the cell types in the human testes. Using single-cell RNA-sequencing (scRNAseq) analysis (10X Genomics platform), we identified 14 different cell types and stages, including 9 germ-cell subsets, in the adult testis. One of these germ-cell subsets appears—based on multiple lines of evidence—to be highly enriched for spermatoginial stem cells (SSCs), the only self-renewing cells in the adult testis. Several novel gene and protein markers labeling this SSC-enriched subset were identified and validated. In neonatal testes (2 and 7 days old), we identified 3 germ cell subsets, including primordial germ cell (PGC) like cells that are closely related to primordial germ cells (PGCs) from human embryos. Using pseudo-developmental trajectory analysis, we assigned cells from the PGCs stage through the PGCL and other neonatal stages to adult SSCs. We also traced the development of somatic cells from the neonatal to adult stages. Together our data delineates the identity of human testicular cells and provides a scheme of development of male germline. The SSC markers we identified hold the potential to be useful for reproductive therapies to treat infertility and testicular cancer.
Monday, November 5, 2018
Presenter: Miguel A. Tam, Ph.D., Senior Manager, Product Realization and Marketing at BioLegend
Title: Simultaneous Proteomics and Transcriptomics: TotalSeq™ and The Future of Single Cell Analysis
Summary: Welcome to the new era of single cell analysis! As personalized medicine and other highly specialized life science and medical applications continue to advance, there is an increasing demand to develop cutting edge technologies.
As such, BioLegend now offers TotalSeq antibodies. These antibody-oligonucleotide conjugates seamlessly integrate with existing protocols (e.g. CITE-seq and REAP-seq) aimed at generating protein and RNA measurement simultaneously, from the same single cell. The olives provide a permanent antibody or cell tag, thanks to the barcode embedded in their sequence.
Presenter: Daniel A. Peiffer, Ph.D., Executive Sales Specialist at Illumina
Illumina Updates: 1) Updates on Illumina’s newest library prep releases to enhance your research! 2) Updates on Illumina’s sequencing system portfolio including the new iSeq100!
Monday, December 3, 2018 CANCELLED
Monday, January 7, 2019
C3-SCS (Single Cell Sequencing) Workshop Group/Social
Monday, January 7, 2019
Sanford Burnahm Prebys Medical Discovery Institute
Fishman Auditorium (attached map, parking is free)
* All are welcome and light hors d'oeuvres/drinks will be served
Speaker/Topic for Discussion are as follows:
Presenter: Dr. Sheng Zhong, Professor of Bioengineering at UCSD
Title: Rainbow-seq: Combining Cell Lineage Tracing with Single-Cell RNA Sequencing in Preimplantation Embryos
Summary: We developed the Rainbow-seq technology to trace cell division history and reveal single-cell transcriptomes. With distinct fluorescent protein genes as lineage markers, Rainbow-seq enables each single-cell RNA sequencing (RNA-seq) experiment to simultaneously decode the lineage marker genes and read single-cell transcriptomes. We triggered lineage tracking in each blastomere at the 2-cell stage, observed microscopically inequivalent contributions of the progeny to the two embryonic poles at the blastocyst stage, and analyzed every single cell at either 4- or 8-cell stage with deep paired-end sequencing of full-length transcripts. Although lineage difference was not marked unequivocally at a single-gene level, it became clear when the transcriptome was analyzed as a whole. Moreover, several groups of novel transcript isoforms with embedded repeat sequences exhibited lineage difference, suggesting a possible link between DNA demethylation and cell fate decision. Rainbow-seq bridged a critical gap between division history and single-cell RNA-seq assays.
Published in iScience 2018