SCORE imaging: specimen in a corrected optical rotational enclosure.

Visual data collection is paramount for the majority of scientific research. The added transparency of the zebrafish (Danio rerio) allows for a greater detail of complex biological research that accompanies seemingly simple observational tools. We developed a visual data analysis and collection approach that takes advantage of the cylindrical nature of the zebrafish allowing for an efficient and effective method for image capture that we call Specimen in a Corrected Optical Rotational Enclosure imaging. To achieve a nondistorted image, zebrafish were placed in a fluorinated ethylene propylene tube with a surrounding optically corrected imaging solution (water). By similarly matching the refractive index of the housing (fluorinated ethylene propylene tubing) to that of the inner liquid and outer liquid (water), distortion was markedly reduced, producing a crisp imagable specimen that is able to be fully rotated 360 degrees. A similar procedure was established for fixed zebrafish embryos using convenient, readily available borosilicate capillaries surrounded by 75% glycerol. The method described here could be applied to chemical genetic screening and other related high-throughput methods within the fish community and among other scientific fields.

[1]  Mark J. Thomas,et al.  Nicotine response genetics in the zebrafish , 2009, Proceedings of the National Academy of Sciences.

[2]  R. Franklin,et al.  Zebrafish myelination: a transparent model for remyelination? , 2008, Disease Models & Mechanisms.

[3]  Torsten Rohlfing,et al.  Towards digital representation of Drosophila embryogenesis , 2008, 2008 5th IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[4]  Michael Brand,et al.  Micro fluid segment technique for screening and development studies on Danio rerio embryos. , 2007, Lab on a chip.

[5]  James Sharpe,et al.  FishNet: an online database of zebrafish anatomy , 2007, BMC Biology.

[6]  Eric W. Klee,et al.  Genome-Wide Reverse Genetics Framework to Identify Novel Functions of the Vertebrate Secretome , 2006, PloS one.

[7]  S. Joshi,et al.  High-throughput zebrafish histology. , 2006, Methods.

[8]  S. Sivasubbu,et al.  Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development , 2006, Mechanisms of Development.

[9]  Leonard I Zon,et al.  Transplantation and in vivo imaging of multilineage engraftment in zebrafish bloodless mutants , 2003, Nature Immunology.

[10]  S. Sivasubbu,et al.  Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon. , 2003, Developmental biology.

[11]  B. Weinstein,et al.  In vivo imaging of embryonic vascular development using transgenic zebrafish. , 2002, Developmental biology.

[12]  D. L. Kerbow,et al.  Physical Constants of Fluoropolymers , 1999 .

[13]  J. Seferis.  Refractive Indices of Polymers , 1999 .

[14]  C. Kimmel,et al.  Stages of embryonic development of the zebrafish , 1995, Developmental dynamics : an official publication of the American Association of Anatomists.

[15]  M. Westerfield.  The zebrafish book : a guide for the laboratory use of zebrafish (Danio rerio) , 1995 .

[16]  R. Budwig.  Refractive index matching methods for liquid flow investigations , 1994 .

[17]  P. Moroz.  Microscopy and Manipulation of Cells in Elastic Capillaries , 1994 .

[18]  J. Gallagher,et al.  Erratum: Refractive index of water and steam as function of wavelength, temperature and density [J. Phys. Chem. Ref. Data 19, 677 (1990)] , 1990 .

[19]  S. Zackson.  Cell clones and segmentation in leech development , 1982, Cell.