Adult fish were briefly anaesthetised with 0.004% MS-222 (Sigma) and imaged with Canon D5MarkII/MACRO 100 (Figs 1 , 3 ). Fish fixed in 4% paraformaldehyde/0.08% glutaraldehyde (Sigma) (Fig. 2 ) were photographed under a Leica MZ1 stereomicroscope. Metamorphic fish were anaesthetised, embedded in low melting point agarose with 4.5 mg/ml ± epinephrine (Sigma) for melanosome contraction, and photographed under a Leica M205 FA stereomicroscope with a Leica DCF300 FX camera using the software LAS V4.1 to allow multifocus images. An illumination was chosen to display iridophores optimally while xanthophore visibility is poor. Photographs were processed in Adobe Photoshop.
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Imaging Techniques for Pigment Analysis
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Other organizations : Max Planck Institute for Developmental Biology
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Variable analysis
independent variables
- Anesthesia concentration (0.004% MS-222)
- Fixation solution (4% paraformaldehyde/0.08% glutaraldehyde)
- Embedding medium (low melting point agarose with ± epinephrine)
- Imaging of adult fish
- Imaging of fixed fish
- Imaging of metamorphic fish
- Canon D5MarkII/MACRO 100 camera
- Leica MZ1 stereomicroscope
- Leica M205 FA stereomicroscope with Leica DCF300 FX camera
- LAS V4.1 software
- Illumination to display iridophores
- Adobe Photoshop for image processing
- Positive control: None specified
- Negative control: None specified
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