PMID- 24562098
OWN - NLM
STAT- MEDLINE
DCOM- 20140721
LR  - 20211021
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 84
DP  - 2014 Feb 7
TI  - Using microfluidics chips for live imaging and study of injury responses in 
      Drosophila larvae.
PG  - e50998
LID - 10.3791/50998 [doi]
LID - 50998
AB  - Live imaging is an important technique for studying cell biological processes, 
      however this can be challenging in live animals. The translucent cuticle of the 
      Drosophila larva makes it an attractive model organism for live imaging studies. 
      However, an important challenge for live imaging techniques is to noninvasively 
      immobilize and position an animal on the microscope. This protocol presents a 
      simple and easy to use method for immobilizing and imaging Drosophila larvae on a 
      polydimethylsiloxane (PDMS) microfluidic device, which we call the 'larva chip'. 
      The larva chip is comprised of a snug-fitting PDMS microchamber that is attached 
      to a thin glass coverslip, which, upon application of a vacuum via a syringe, 
      immobilizes the animal and brings ventral structures such as the nerve cord, 
      segmental nerves, and body wall muscles, within close proximity to the coverslip. 
      This allows for high-resolution imaging, and importantly, avoids the use of 
      anesthetics and chemicals, which facilitates the study of a broad range of 
      physiological processes. Since larvae recover easily from the immobilization, 
      they can be readily subjected to multiple imaging sessions. This allows for 
      longitudinal studies over time courses ranging from hours to days. This protocol 
      describes step-by-step how to prepare the chip and how to utilize the chip for 
      live imaging of neuronal events in 3(rd) instar larvae. These events include the 
      rapid transport of organelles in axons, calcium responses to injury, and 
      time-lapse studies of the trafficking of photo-convertible proteins over long 
      distances and time scales. Another application of the chip is to study 
      regenerative and degenerative responses to axonal injury, so the second part of 
      this protocol describes a new and simple procedure for injuring axons within 
      peripheral nerves by a segmental nerve crush.
FAU - Mishra, Bibhudatta
AU  - Mishra B
AD  - Department of Molecular, Cellular and Developmental Biology, University of 
      Michigan.
FAU - Ghannad-Rezaie, Mostafa
AU  - Ghannad-Rezaie M
AD  - Department of Biomedical Engineering, University of Michigan.
FAU - Li, Jiaxing
AU  - Li J
AD  - Department of Molecular, Cellular and Developmental Biology, University of 
      Michigan.
FAU - Wang, Xin
AU  - Wang X
AD  - Department of Molecular, Cellular and Developmental Biology, University of 
      Michigan.
FAU - Hao, Yan
AU  - Hao Y
AD  - Department of Molecular, Cellular and Developmental Biology, University of 
      Michigan.
FAU - Ye, Bing
AU  - Ye B
AD  - Life Sciences Institute, University of Michigan; Department of Cell and 
      Developmental Biology, University of Michigan.
FAU - Chronis, Nikos
AU  - Chronis N
AD  - Department of Biomedical Engineering, University of Michigan; Department of 
      Mechanical Engineering, University of Michigan.
FAU - Collins, Catherine A
AU  - Collins CA
AD  - Department of Molecular, Cellular and Developmental Biology, University of 
      Michigan; collinca@umich.edu.
LA  - eng
GR  - R00MH080599/MH/NIMH NIH HHS/United States
GR  - R00 MH080599/MH/NIMH NIH HHS/United States
GR  - R21 NS062313/NS/NINDS NIH HHS/United States
GR  - NS069844/NS/NINDS NIH HHS/United States
GR  - R01 NS069844/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Video-Audio Media
DEP - 20140207
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Dimethylpolysiloxanes)
RN  - 63148-62-9 (baysilon)
SB  - IM
MH  - Animals
MH  - Dimethylpolysiloxanes
MH  - Drosophila melanogaster
MH  - Larva
MH  - Microfluidic Analytical Techniques/instrumentation/*methods
MH  - Neurons/*cytology
PMC - PMC4117361
EDAT- 2014/02/25 06:00
MHDA- 2014/07/22 06:00
PMCR- 2015/02/07
CRDT- 2014/02/25 06:00
PHST- 2014/02/25 06:00 [entrez]
PHST- 2014/02/25 06:00 [pubmed]
PHST- 2014/07/22 06:00 [medline]
PHST- 2015/02/07 00:00 [pmc-release]
AID - 50998 [pii]
AID - 10.3791/50998 [doi]
PST - epublish
SO  - J Vis Exp. 2014 Feb 7;(84):e50998. doi: 10.3791/50998.