QBio Seminar - Single-cell analysis with RNA sequencing, quantitative imaging, and microfluidics

Event Date: 

Monday, February 2, 2015 - 11:00am

Event Location: 

  • KITP Auditorium

Event Contact: 

Speaker: Dr. Aaron Streets of Peking University, Beijing China

 

Abstract:

Single-cell analysis enables measurement of biological variation in heterogeneous cellular populations and dissection of phenotypic complexity that is masked in ensemble measurement. This seminar presents recent advances combining next generation sequencing, microscopy, and microfluidic technology for characterization of phenotypic heterogeneity at the single-cell level. We developed a microfluidic approach to prepare cDNA from single cells for high-throughput transcriptome sequencing. The microfluidic platform facilitates single-cell manipulation, minimizes contamination, and furthermore, provides improved detection sensitivity and measurement precision, which is necessary for differentiating biological variability from technical noise. Many cellular characteristics, however, are not directly genetically encoded, and therefore cannot be assessed with genomic tools. For example, variable metabolism of fatty acid leads to heterogeneity in lipid droplet content within cells from the same population. In such cases quantitative microscopy is a valuable tool for measuring morphological variation between single cells. I will present a microfluidic cell culture platform that facilitates systematic, quantitative imaging of intercellular lipid droplets in single cells using stimulated Raman scattering microscopy.  This microfluidic cell culture array enables parallel screening and high-throughput imaging of cellular response to exogenous fatty acids, while quantitative image processing allows for morphological analysis at the single-cell level. Finally, by combining single-cell RNA sequencing with microscopy, quantitative connections can be made between gene expression and morphological heterogeneity in single cells.