{"CIL_CCDB":{"Status":{"Is_public":true,"Deleted":false,"Publish_time":1658479140},"Data_type":{"Still_image":false,"Z_stack":true,"Video":false,"Time_series":false},"CIL":{"CORE":{"TECHNICALDETAILS":{"free_text":"Brief description of sample preparation and SBEM imaging\r\n<br><br>\r\n5 d.p.f. zebrafish larva was prepared for SBEM. Larva was fixed in 2.5% glutaraldehyde and 4% paraformaldehyde in 0.15 M cacodylate buffer (CB, pH 7.4) at 4°C overnight. After removing the fixative, larva was treated for 15 minutes with 20 mM glycine in 0.15M CB, on ice to quench the unreacted glutaraldehyde. Then, larva was washed with 0.15 M CB and then placed into 2% OsO4\/1.5% potassium ferrocyanide in 0.15 M CB containing 2 mM CaCl2. The larva was left for 30 min on ice and then 30 min at room temperature (RT). After thorough washing in double distilled water (ddH2O), larva was placed into 0.05% thiocarbohydrazide for 30 min. Larva was again washed and then stained with 2% aqueous OsO4 for 30 min. Larva was washed and then placed into 2% aqueous uranyl acetate overnight at 4°C. Larva was washed with ddH2O at RT and then stained with 0.05% en bloc lead aspartate for 30 min at 60°C. Larva was washed with ddH2O and then dehydrated on ice in 50%, 70%, 90%, 100%, 100% ethanol solutions for 10 min at each step. Larva was then washed twice with dry acetone and placed into 50:50 Durcupan ACM:acetone overnight. Larva was transferred to 100% Durcupan resin overnight. Larva was then flat embedded between glass slides coated with mould-release compound and left in an oven at 60°C for 72 h.  The larva block was a Zeiss Gemini scanning electron microscope equipped with a Gatan 3View2XP and OnPoint backscatter detector. Images were acquired at 4.0 kV accelerating voltage with a 30 μm condenser aperture and 0.5 μsec dwell time; Z step size was 70 nm; pixel size was 9.5 nm; raster size was 25k x 12k, variable pressure under nitrogen. The volume was aligned using cross correlation, segmented, and visualized using IMOD."},"TERMSANDCONDITIONS":{"free_text":"attribution_nc_sa"},"ATTRIBUTION":{"Contributors":["Keun-Young Kim","Sobhika Agarwala","Sebastien Phan","Saeyeon Ju","Ye Eun Kong","Guillaume A. Castillon","Eric A. Bushong","Mark H. Ellisman","Owen J. Tamplin"]},"IMAGINGMODE":[{"free_text":"SBEM"}],"IMAGEDESCRIPTION":{"free_text":"Transverse sections of  5 days post-fertilization wild-type zebrafish larva in the region of the anterior  kidney"},"NCBIORGANISMALCLASSIFICATION":[{"onto_name":"Danio rerio","onto_id":"NCBITaxon:7955"}],"CELLTYPE":[{"free_text":"Whole larva"}],"GROUP_ID":"20095"},"Image_files":[{"File_type":"Jpeg","File_path":"54850.jpg","Size":309231,"Mime_type":"image\/jpeg; charset=utf-8"},{"File_type":"Zip","File_path":"54850.zip","Size":302177,"Mime_type":"application\/zip"}],"Alternative_image_files":[{"File_type":"Mrc","URL_postfix":"\/media\/images\/54850\/Conv-Pb-ZFish-II-3view-VP-bin1-8bit.mrc","File_path":"Conv-Pb-ZFish-II-3view-VP-bin1-8bit.mrc","Size":460298513258,"Mime_type":"application\/octet-stream"}]},"Citation":{"Title":"Keun-Young Kim;Sobhika Agarwala;Sebastien Phan;Saeyeon Ju;Ye Eun Kong;Guillaume A. Castillon;Eric A. Bushong;Mark H. Ellisman;Owen J. Tamplin (2022) CIL:54850, Danio rerio, Whole larva. CIL. Dataset. CIL. Dataset","ARK":"ark:\/b7295\/w9cil54850","DOI":"doi:10.7295\/W9CIL54850"}}}