|Title:||Two Z stacks from the coelomic development of the planktotrophic larva of the sea urchin Heliocidaris tuberculata recorded by high resolution confocal laser scanning microscopy, published in Morris et al. (2018)|
|Authors:||Morris, Valerie B.|
|Abstract:||The Z stacks are those from which figure 2B,C of a 24 hour larva and figure 5A-E and 5G,H of a 55 hour larva in Morris et al. (2018) were constructed. The Z stack of figure 2B,C shows cells at the head of the archenteron where the coelomic pouches and the oesophagus form; cells with large profiles, possible progeny of the small micromeres, lie between the left and right coelomic pouches. Figure 5 shows the coelomic pouch growth zone.|
|Description:||Morris et al. (2018) describes coelomic development in Heliocidaris tuberculata, which is a sea urchin that develops indirectly and which is the congener of the sea urchin Heliocidaris erythrogramma that develops directly. The key difference between indirect and direct development is the formation in indirect development of four micromeres at the vegetal pole of the zygote at the fourth cell division, from which small micromeres form at subsequent cell divisions. The development of the assumed progeny of the small micromeres is described. The progeny appear to be stem cells that build anlagen of the planktotrophic, feeding larva and the adult sea urchin of indirect development.
Adult H. tuberculata were collected from coastal waters of New South Wales, Australia. Larvae from fertilizations were cultured at 24 ˚C in filtered sea water (FSW). They were fixed in 2.5% (v/v) glutaraldehyde/FSW for 1-2 h, washed in FSW, then dehydrated to 70% (v/v) in graded ethanols/Millepore filtered water (MFW) and stored at 4 ˚C. For viewing, the larvae were further dehydrated to 100% (v/v) in graded ethanols/MFW, then cleared and mounted in 2:1 (v/v) methyl benzoate/methyl alcohol, in cavity slides.
Larvae, autofluorescent from the glutaraldehyde fixation, were viewed in the Leica TCS SP5 MP multi-photon laser scanning confocal system (Leica Microsystems) with a tunable Mai Tai Deep See laser (Spectra-Physics) attached to a Leica DMI6000B-CS inverted microscope. Each larva was imaged using multi-photon microscopy at ex = 870 nm with pulses in the 100-200 femtosecond range and detected in a reflected non-descanned detector at em = 545-605 nm. A Z stack was collected, with default X flipped, averaged over two frames in a 1024 x 1024 pixel array, 12 bits/pixel, at a slice thickness of 0.5 m using a Leica HCX PL APO 63x/1.30 GLYC CORR CS 21˚C objective lens.
Each Z stack was saved as a .lif file by the SP5 Leica Microsystems software. The .lif files were each exported as a folder of .tif files with the .lif metadata deleted. The filenames of the folders are Fig. 2B,C and Fig. 5A-E and 5G,H, which are the names of the figures in Morris et al. (2018).
Each .tif image can be viewed individually or the series of .tif images contained in a folder can be analysed by image analysis software such as ImageJ (version 1.43r) available from the National Institutes of Health site (http://rsbweb.nih.gov/ij/).|
Associated publication Morris VB, Kable E, Koop D, Cisternas P, Byrne M. (2018) Stem cells in a sea urchin larva exhibit morphogenetic capabilities. (submitted for publication)
|Type of Work:||Dataset|
|Type of Publication:||Publisher version|
|Appears in Collections:||Research Papers and Publications. Biological Sciences|
|Fig_2BC.zip||Folder of Z stack .tif files compressed (Fig. 2B,C)||299.13 MB||Compressed (zipped) Folder (.zip)|
|Fig_5AE_5GH.zip||Folder of Z stack .tif files compressed (Fig. 5A-E and 5G,H)||374.98 MB||Compressed (zipped) Folder (.zip)|