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http://hdl.handle.net/2123/7028
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| Title: | The Z stack of a vestibula larva of the sea urchin Heliocidaris erythrogramma recorded by confocal laser scanning microscopy |
| Authors: | Morris, Valerie B School of Biological Sciences |
| Keywords: | serial imaging morphogenesis echinoderm evolution |
| Issue Date: | 23-Nov-2010 |
| Abstract: | The Z stack in this archive is representative of a vestibula larva of H. erythrogramma at an early stage in the development of the hydrocoele recorded in an Olympus FluoView 1000 laser scanning system. It is from a study of an echinoid with abbreviated larval development that shows the morphogenesis of the adult coeloms. The development is asymmetric with a larger left coelom forming on the larval-left side and a smaller right coelom forming on the larval-right side. The hydrocoele forms from the anterior part of the left coelom. The five lobes of the hydrocoele from which the pentamery of the adult derives take shape on the outer distal wall of the anterior part of the left coelom. The hydrocoele separates from the more posterior part of the left coelom, which becomes the left posterior coelom. Both larval and adult polarities are evident in this larva, supporting an origin from a single ancestral polarity. |
| Description: | The larva was fixed 36 hours after fertilization from a culture maintained at 20˚C. It was fixed in 2.5% (v/v) glutaraldehyde in filtered sea water for 1-2 hours, dehydrated in an ethanol series to 100% ethanol, then cleared in 2:1 (v/v) benzyl benzoate/benzyl alcohol. The larva, made autofluorescent by the glutaraldehyde fixation, was viewed in the clearant after mounting in a coverslip-sealed chamber. It was viewed in an Olympus FluoView 1000 laser scanning system (version 1.7.1.0) linked to an Olympus IX81 inverted microscope. The larva was excited at λex = 633 nm with a helium-neon laser and detected at λem = 645-745 nm. The Z stack was collected at a voxel dimension of 1.22 x 1.22 x 1.41 μm averaged over two frames in a 512 x 512 pixel array 12 bits/pixel using a 20x/NA 0.7 UplanApo objective lens. The orientation of the larva in the Z stack is such that view is of the larval-left side with larval posterior on the left in each image. To obtain a corrected orientation, as shown in the single tif image, the images should be rotated and also reflected, since the images in the Z stack were reflected by the IX81 inverted microscope. Each tif image can be viewed individually or the series of tif images contained in the folder can be analysed by image analysis software such as ImageJ available from the National Institutes of Health site (http://rsbweb.nih.gov/ij/). The larva is in frontal view with respect to the adult coeloms, providing an oral view of the hydrocoele and its five lobes. Associated publication Morris VB (2011) Coelomogenesis during the abbreviated development of the echinoid Heliocidaris erythrogramma and the developmental origin of the echinoderm pentameral body plan. (submitted for publication). |
| URI: | http://hdl.handle.net/2123/7028 |
| Department/Unit/Centre: | School of Biological Sciences |
| Appears in Collections: | Research Papers and Publications. Biological Sciences |
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