zWEDGI: Wounding and Entrapment Device for Imaging Live Zebrafish Larvae.
Zebrafish, an established model organism in developmental biology, is also a valuable tool for imaging wound healing in space and time with cellular resolution. However, long-term imaging of wound healing poses technical challenges as wound healing occurs over multiple temporal scales. The traditional strategy of larval encapsulation in agarose successfully limits sample movement but impedes larval development and tissue regrowth and is therefore not amenable to long-term imaging of wound healing. To overcome this challenge, we engineered a functionally compartmentalized device, the zebrafish Wounding and Entrapment Device for Growth and Imaging (zWEDGI), to orient larvae for high-resolution microscopy, including confocal and second harmonic generation (SHG), while allowing unrestrained tail development and regrowth. In this device, larval viability was maintained and tail regrowth was improved over embedding in agarose. The quality of tail fiber SHG images collected from larvae in the device was similar to fixed samples but provided the benefit of time lapse data collection. Furthermore, we show that this device was amenable to long-term (>24 h) confocal microscopy of the caudal fin. Finally, the zWEDGI was designed and fabricated using readily available techniques so that it can be easily modified for diverse experimental imaging protocols.
1 Morgridge Institute for Research , Madison, Wisconsin.|2 Department of Biomedical Engineering, UW-Madison , Madison, Wisconsin.|3 Laboratory for Optical and Computational Instrumentation, UW-Madison , Madison, Wisconsin.
Kayla
K
Huemer
3 Laboratory for Optical and Computational Instrumentation, UW-Madison , Madison, Wisconsin.
Jayne M
JM
Squirrell
1 Morgridge Institute for Research , Madison, Wisconsin.
Robert
R
Swader
4 Cellular and Molecular Pathology Graduate Program, UW-Madison , Madison, Wisconsin.
Danny C
DC
LeBert
5 Department of Medical Microbiology and Immunology.|6 Department of Pediatrics, UW-Madison , Madison, Wisconsin.
Anna
A
Huttenlocher
1 Morgridge Institute for Research , Madison, Wisconsin.|2 Department of Biomedical Engineering, UW-Madison , Madison, Wisconsin.|3 Laboratory for Optical and Computational Instrumentation, UW-Madison , Madison, Wisconsin.
Kevin W
KW
Eliceiri
D000818:Animals; D015509:Developmental Biology / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007814:Larva / Q000502:physiology; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods; D014945:Wound Healing; D015027:Zebrafish / Q000254:growth & development / Q000502:physiology*
device; live imaging; microscopy; wounding
Development. 2015 Jun 15;142(12):2136-46
26015541
Nat Biotechnol. 1999 Aug;17(8):763-7
10429240
Methods Cell Biol. 2016;133:55-68
27263408
Ostomy Wound Manage. 2006 Mar;52(3):26-8, 30, 32 passim
16567857
Plast Reconstr Surg. 2006 Jun;117(7 Suppl):59S-71S
16799375
IEEE Trans Image Process. 1998;7(1):27-41
18267377
Fish Shellfish Immunol. 2008 Oct;25(4):341-50
18640057
Ostomy Wound Manage. 2005 May;51(5):38-54; quiz 55-6
16014984
Biophys J. 2002 Jan;82(1 Pt 1):493-508
11751336
Zebrafish. 2010 Jun;7(2):149-54
20528262
Cell Biochem Biophys. 2009;53(3):145-57
19259625
Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1271-5
1741380
Dev Biol. 2011 Jun 1;354(1):160-72
21420398
Cell Microbiol. 2016 Apr;18(4):591-604
26468080
Infect Immun. 2013 Jan;81(1):110-21
23090960
J Embryol Exp Morphol. 1985 Jun;87:145-61
4031750
Adv Wound Care (New Rochelle). 2015 Sep 1;4(9):560-582
26339534
Pathol Int. 1994 Feb;44(2):96-106
7517762
Biophys J. 1998 Oct;75(4):2015-24
9746543
Lab Chip. 2013 May 7;13(9):1732-6
23503983
Dev Biol. 2002 Aug 15;248(2):307-18
12167406
Nat Methods. 2012 Jun 28;9(7):676-82
22743772
Int J Biochem Cell Biol. 2004 Jun;36(6):1031-7
15094118
Neural Dev. 2011 May 27;6:27
21619654
Proc Natl Acad Sci U S A. 2002 Jun 25;99(13):8713-8
12060710
Dev Comp Immunol. 2008;32(7):745-57
18222541
J Cell Biol. 2012 Oct 15;199(2):225-34
23045550
Dev Dyn. 2004 Dec;231(4):693-9
15499559
Endocrinology. 2002 Jan;143(1):47-54
11751590
Nature. 2009 Jun 18;459(7249):921-3
19536251