Long-term C. elegans immobilization enables high resolution developmental studies in vivo

被引:28
作者
Berger, Simon [1 ]
Lattmann, Evelyn [2 ]
Aegerter-Wilmsen, Tinri [2 ]
Hengartner, Michael [2 ]
Hajnal, Alex [2 ]
deMello, Andrew [1 ]
Casadevall i Solvas, Xavier [1 ]
机构
[1] Swiss Fed Inst Technol, Inst Chem & Bioengn, CH-8093 Zurich, Switzerland
[2] Univ Zurich, Inst Mol Life Sci, CH-8057 Zurich, Switzerland
基金
瑞士国家科学基金会;
关键词
CAENORHABDITIS-ELEGANS; ON-CHIP; CELL-MIGRATION; DEATH; MICROFLUIDICS; GERMLINE; INVASION; NEMATODE; WORMS;
D O I
10.1039/c7lc01185g
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Live-imaging of C. elegans is essential for the study of conserved cellular pathways (e.g. EGFR/Wnt signaling) and morphogenesis in vivo. However, the usefulness of live imaging as a research tool has been severely limited by the need to immobilize worms prior to and during imaging. Conventionally, immobilization is achieved by employing both physical and chemical interventions. These are known to significantly affect many physiological processes, and thus limit our understanding of dynamic developmental processes. Herein we present a novel, easy-to-use microfluidic platform for the long-term immobilization of viable, normally developing C. elegans, compatible with image acquisition at high resolution, thereby overcoming the limitations associated with conventional worm immobilization. The capabilities of the platform are demonstrated through the continuous assessment of anchor cell (AC) invasion and distal tip cell (DTC) migration in larval C. elegans and germ cell apoptosis in adult C. elegans in vivo for the first time.
引用
收藏
页码:1359 / 1368
页数:10
相关论文
共 37 条
[1]   Ways of dying: multiple pathways to apoptosis [J].
Adams, JM .
GENES & DEVELOPMENT, 2003, 17 (20) :2481-2495
[2]   Control of cell migration during Caenorhabditis elegans development [J].
Blelloch, R ;
Newman, C ;
Kimble, J .
CURRENT OPINION IN CELL BIOLOGY, 1999, 11 (05) :608-613
[3]   Live imaging of cellular dynamics during Caenorhabditis elegans postembryonic development [J].
Chai, Yongping ;
Li, Wei ;
Feng, Guoxin ;
Yang, Yihong ;
Wang, Xiangming ;
Ou, Guangshuo .
NATURE PROTOCOLS, 2012, 7 (12) :2090-2102
[4]   Why Caenorhabditis elegans adults sacrifice their bodies to progeny [J].
Chen, JJ ;
Caswell-Chen, EP .
NEMATOLOGY, 2003, 5 :641-645
[5]   CO2 and compressive immobilization of C. elegans on-chip [J].
Chokshi, Trushal Vijaykumar ;
Ben-Yakar, Adela ;
Chronis, Nikos .
LAB ON A CHIP, 2009, 9 (01) :151-157
[6]   Microfluidics for in vivo imaging of neuronal and behavioral activity in Caenorhabditis elegans [J].
Chronis, Nikos ;
Zimmer, Manuel ;
Bargmann, Cornelia I. .
NATURE METHODS, 2007, 4 (09) :727-731
[7]   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
[8]   Microfluidic chamber arrays for whole-organism behavior-based chemical screening [J].
Chung, Kwanghun ;
Zhan, Mei ;
Srinivasan, Jagan ;
Sternberg, Paul W. ;
Gong, Emily ;
Schroeder, Frank C. ;
Lu, Hang .
LAB ON A CHIP, 2011, 11 (21) :3689-3697
[9]  
Crane MM, 2012, NAT METHODS, V9, P977, DOI [10.1038/NMETH.2141, 10.1038/nmeth.2141]
[10]   Cell death: Critical control points [J].
Danial, NN ;
Korsmeyer, SJ .
CELL, 2004, 116 (02) :205-219