Automated on-chip rapid microscopy, phenotyping and sorting of C. elegans.
Microscopy, phenotyping and visual screens are frequently applied to model organisms in combination with genetics. Although widely used, these techniques for multicellular organisms have mostly remained manual and low-throughput. Here we report the complete automation of sample handling, high-resolution microscopy, phenotyping and sorting of Caenorhabditis elegans. The engineered microfluidic system, coupled with customized software, has enabled high-throughput, high-resolution microscopy and sorting with no human intervention and may be combined with any microscopy setup. The microchip is capable of robust local temperature control, self-regulated sample-loading and automatic sample-positioning, while the integrated software performs imaging and classification of worms based on morphological and intensity features. We demonstrate the ability to perform sensitive and quantitative screens based on cellular and subcellular phenotypes with over 95% accuracy per round and a rate of several hundred worms per hour. Screening time can be reduced by orders of magnitude; moreover, screening is completely automated.
Chung
Kwanghun
K
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, 311 Ferst Dr. NW, Atlanta, Georgia 30332, USA.
Crane
Matthew M
MM
Lu
Hang
H
D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000166:cytology* / Q000235:genetics*; D004867:Equipment Design; D020869:Gene Expression Profiling / Q000379:methods; D017238:Genes, Helminth; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data; D044085:Microfluidics; D008853:Microscopy / Q000379:methods; D009154:Mutation; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D011994:Recombinant Proteins / Q000235:genetics; D012984:Software; D013569:Synapses / Q000378:metabolism
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins