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morphometric image analysis  (Oxford Instruments)


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    Oxford Instruments morphometric image analysis
    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of <t>morphometric</t> analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.
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    Images

    1) Product Images from "Label-free multiphoton imaging reveals volumetric shifts across development in sensory-related brain regions of a miniature transparent vertebrate"

    Article Title: Label-free multiphoton imaging reveals volumetric shifts across development in sensory-related brain regions of a miniature transparent vertebrate

    Journal: bioRxiv

    doi: 10.1101/2024.07.18.604134

    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of morphometric analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.
    Figure Legend Snippet: Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of morphometric analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.

    Techniques Used: Imaging, Transformation Assay



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    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of <t>morphometric</t> analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.
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    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of <t>morphometric</t> analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.
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    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of <t>morphometric</t> analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.
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    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of <t>morphometric</t> analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.
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    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of <t>morphometric</t> analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.
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    universal imaging inc computer-based morphometric analysis image 1
    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of <t>morphometric</t> analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.
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    Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of morphometric analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.

    Journal: bioRxiv

    Article Title: Label-free multiphoton imaging reveals volumetric shifts across development in sensory-related brain regions of a miniature transparent vertebrate

    doi: 10.1101/2024.07.18.604134

    Figure Lengend Snippet: Third harmonic generation (THG) brain imaging in groups of Danionella dracula 3 to 90 dpf. A . Left: Side view of a male. Scale bar indicates 3mm. Right: Head-on view of male extending its hypertrophied lower jaw during mid-lunge at another male. B . Dorsal views of fish imaged from 90-3 dpf (left to right). Scale bar left indicates 500 μm for all images. Top insert at 90 dpf is zoom-in on fangs (indicated by arrows; dashed outline on left fang), which are first apparent in 90 dpf males along with hypertrophied lower jaw. C . Schematic of imaging apparatus (see Materials and Methods). Left: profile drawing of animal placement under objective. Right: dorsal view of adult placed in putty holder with perfusion mouthpiece. Fish imaged less than 60 dpf were imaged using an alternative setup than pictured (see Materials and Methods, Fish stabilization for imaging). D. 30, 21, and 14 dpf example fish depicting changes in swim bladder (SB) separation into anterior and posterior chambers. Scale bar indicates 500 μm for all images. E . Schematic of representative D. dracula brain; illustrations based on pictures of dissected whole brains. Brain regions of morphometric analysis indicated in lateral and dorsal views; green, OB (olfactory bulb); blue, telencephalon; orange, TL (torus longitudinalis); pink, TeO (optic tectum). Insert indicates horizontal slice in midbrain depicting TeO (pink), periventricular gray zone (PGZ) of TeO (purple, included in TeO measurement), depicted for indication of boundary between TeO and TS (yellow, torus semicircularis). F . Scatterplot of age in dpf of animals in this study and gross brain volume in μm 3 . Boxplots indicate median, upper and lower quartiles and 1.5× interquartile range for each Age. Gross brain volume calculation is described in and Materials and Methods. Different ages depicted by different colored dots as shown in legend: red, 5 dpf; yellow, 14 dpf; green, 21 dpf; teal, 30 dpf; blue, 60 dpf; purple, 90 dpf. As is described in Materials and Methods, we performed the best fitting quadratic regression (F 2,30= 120.8, R 2 = 0.8821, p<0.001) on log-transformed data to normalize the residuals. The green line displayed here represents the quadratic regression line based on the original (non-transformed) data, for best visual clarity and understanding.

    Article Snippet: Morphometric image analysis was performed using the software Imaris (BITPLANE, Oxford Instruments).

    Techniques: Imaging, Transformation Assay