An automated microfluidic platform for C-elegans embryo arraying, phenotyping, and long-term live imaging

被引:49
作者
Cornaglia, Matteo [1 ]
Mouchiroud, Laurent [2 ]
Marette, Alexis [1 ]
Narasimhan, Shreya [1 ]
Lehnert, Thomas [1 ]
Jovaisaite, Virginija [2 ]
Auwerx, Johan [2 ]
Gijs, Martin A. M. [1 ]
机构
[1] Ecole Polytech Fed Lausanne, Lab Microsyst, CH-1015 Lausanne, Switzerland
[2] Ecole Polytech Fed Lausanne, Lab Integrat & Syst Physiol, CH-1015 Lausanne, Switzerland
基金
瑞士国家科学基金会;
关键词
UNFOLDED PROTEIN RESPONSE; CAENORHABDITIS-ELEGANS; MITOCHONDRIAL UPR; GENE-EXPRESSION; ACTIVATION; LONGEVITY; PATHWAY;
D O I
10.1038/srep10192
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Studies of the real-time dynamics of embryonic development require a gentle embryo handling method, the possibility of long-term live imaging during the complete embryogenesis, as well as of parallelization providing a population's statistics, while keeping single embryo resolution. We describe an automated approach that fully accomplishes these requirements for embryos of Caenorhabditis elegans, one of the most employed model organisms in biomedical research. We developed a microfluidic platform which makes use of pure passive hydrodynamics to run on-chip worm cultures, from which we obtain synchronized embryo populations, and to immobilize these embryos in incubator microarrays for long-term high-resolution optical imaging. We successfully employ our platform to investigate morphogenesis and mitochondrial biogenesis during the full embryonic development and elucidate the role of the mitochondrial unfolded protein response (UPRmt) within C. elegans embryogenesis. Our method can be generally used for protein expression and developmental studies at the embryonic level, but can also provide clues to understand the aging process and age-related diseases in particular.
引用
收藏
页数:13
相关论文
共 34 条
[1]   Miniaturized Embryo Array for Automated Trapping, Immobilization and Microperfusion of Zebrafish Embryos [J].
Akagi, Jin ;
Khoshmanesh, Khashayar ;
Evans, Barbara ;
Hall, Chris J. ;
Crosier, Kathryn E. ;
Cooper, Jonathan M. ;
Crosier, Philip S. ;
Wlodkowic, Donald .
PLOS ONE, 2012, 7 (05)
[2]  
Albrecht DR, 2011, NAT METHODS, V8, P599, DOI [10.1038/NMETH.1630, 10.1038/nmeth.1630]
[3]   Microfluidic laboratories for C. elegans enhance fundamental studies in biology [J].
Bakhtina, Natalia A. ;
Korvink, Jan G. .
RSC ADVANCES, 2014, 4 (09) :4691-4709
[4]   Microfluidics for the analysis of behavior, nerve regeneration, and neural cell biology in C. elegans [J].
Ben-Yakar, Adela ;
Chronis, Nikos ;
Lu, Hang .
CURRENT OPINION IN NEUROBIOLOGY, 2009, 19 (05) :561-567
[5]   Ubiquitin-like protein 5 positively regulates chaperone gene expression in the mitochondrial unfolded protein response [J].
Benedetti, Cristina ;
Haynes, Cole M. ;
Yang, Yun ;
Harding, Heather P. ;
Ron, David .
GENETICS, 2006, 174 (01) :229-239
[6]  
Chisholm Andrew D., 2005, Epidermal Morphogenesis
[7]   Worm chips: Microtools for C. elegans biology [J].
Chronis, Nikos .
LAB ON A CHIP, 2010, 10 (04) :432-437
[8]   Automated on-chip rapid microscopy, phenotyping and sorting of C. elegans [J].
Chung, Kwanghun ;
Crane, Matthew M. ;
Lu, Hang .
NATURE METHODS, 2008, 5 (07) :637-643
[9]   A microfluidic array for large-scale ordering and orientation of embryos [J].
Chung, Kwanghun ;
Kim, Yoosik ;
Kanodia, Jitendra S. ;
Gong, Emily ;
Shvartsman, Stanislav Y. ;
Lu, Hang .
NATURE METHODS, 2011, 8 (02) :171-U103
[10]   Lifespan regulation by evolutionarily conserved genes essential for viability [J].
Curran, Sean P. ;
Ruvkun, Gary .
PLOS GENETICS, 2007, 3 (04) :0479-0487