PUBLICATION

Microfluidic device for a rapid immobilization of Zebrafish larvae in environmental scanning electron microscopy

Authors
Akagi, J., Zhu, F., Skommer, J., Hall, C.J., Crosier, P.S., Cialkowski, M., Wlodkowic, D.
ID
ZDB-PUB-141209-15
Date
2015
Source
Cytometry. Part A : the journal of the International Society for Analytical Cytology   87(3): 190-4 (Other)
Registered Authors
Crosier, Phil, Hall, Chris
Keywords
Danio rerio, Zebrafish, environmental scanning electron microscopy, imaging, immobilization, lab-on-a-chip, larvae, microfluidics
MeSH Terms
  • Animals
  • Cells, Immobilized/ultrastructure*
  • Lab-On-A-Chip Devices*
  • Larva
  • Microfluidic Analytical Techniques/methods*
  • Microscopy, Electron, Scanning/methods
  • Time Factors
  • Zebrafish*
PubMed
25483307 Full text @ Cytometry A
Abstract
Small vertebrate model organisms have recently gained popularity as attractive experimental models that enhance our understanding of human tissue and organ development. Despite a large body of evidence using optical spectroscopy for the characterization of small model organism on chip-based devices, no attempts have been so far made to interface microfabricated technologies with environmental scanning electron microscopy (ESEM). Conventional scanning electron microscopy requires high vacuum environments and biological samples must be, therefore, submitted to many preparative procedures to dehydrate, fix, and subsequently stain the sample with gold-palladium deposition. This process is inherently low-throughput and can introduce many analytical artifacts. This work describes a proof-of-concept microfluidic chip-based system for immobilizing zebrafish larvae for ESEM imaging that is performed in a gaseous atmosphere, under low vacuum mode and without any need for sample staining protocols. The microfabricated technology provides a user-friendly and simple interface to perform ESEM imaging on zebrafish larvae. Presented lab-on-a-chip device was fabricated using a high-speed infrared laser micromachining in a biocompatible poly(methyl methacrylate) thermoplastic. It consisted of a reservoir with multiple semispherical microwells designed to hold the yolk of dechorionated zebrafish larvae. Immobilization of the larvae was achieved by a gentle suction generated during blotting of the medium. Trapping region allowed for multiple specimens to be conveniently positioned on the chip-based device within few minutes for ESEM imaging.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes