Andy Miller, Sarah Webb (2011) CIL:12816, Danio rerio. CIL. Dataset

Three calcium waves propagating around the blastodisc margin. This sequence covers 28 min of development around 90% epiboly.The movie represents a 30-sec integration window moving in 5-sec steps, running at 50x real time. Fertilized zebrafish eggs were collected within 5 min of spawning, enzymatically dechorionated, and injected with approximately 0.9 nl of a 1% solution of recombinant f-aequorin in 100 mM KCl, 5 mM Mops, and 50 μM EDTA. During imaging the embryos were maintained at 28°C in 30% Danieau's medium containing penicillin (0.5 mg/ml), streptomycin (5,000 units/ml; Sigma), and 0.5% methylcellulose. Imaging was performed on a Photon Imaging Microscope (Science Wares, Falmouth, MA) that used a photon-counting spatial detector with a resistive anode output (Photek, St. Albens-on-Sea, U.K.). Digitized detector output in the form of a stream of time-labeled eight-bit x--y coordinates (256 × 256 pixels) was used to construct time-lapse imaging sequences. The imaging system software allowed the original photon data stream to be analyzed according to any chosen integration time, with the resulting image frames maintaining accurate photon quantitation up to 256 photons per pixel. These images are part of an image series within the Zebrafish--The Living Laboratory CD made available by Mark Cooper and described in Methods in Cell Biology Volume 77, 2004, Pages 439-457. This is a supplemental video in PNAS January 5, 1999 vol. 96: 157-161.

在胚胎发育过程中，观察到了三次围绕胚盘边缘传播的钙波。本序列覆盖了约28分钟的开发过程，此时胚盘的展开程度达到90%。该视频展示了一个30秒的积分窗口，以5秒为步长进行移动，以50倍实时速度运行。受精斑马鱼卵在产卵后5分钟内收集，通过酶解去卵黄囊，并注入约0.9 nl的1%重组f-aequorin溶液（含100 mM氯化钾，5 mM Mops，以及50 μM EDTA）。在成像过程中，胚胎被维持在28°C的温度下，在含有青霉素（0.5 mg/ml）、链霉素（5,000单位/ml；Sigma）和0.5%羧甲基纤维素钠的30% Danieau's培养液中。成像在Photon Imaging Microscope（Science Wares，Falmouth，MA）上进行，该显微镜使用具有电阻阳极输出的光子计数空间探测器（Photek，St. Albans-on-Sea，U.K.）。以时间标记的八位x-y坐标流（256 × 256像素）的形式数字化探测器输出，用于构建时间推移成像序列。成像系统软件允许根据所选的积分时间分析原始光子数据流，生成的图像帧保持了每像素高达256光子的准确光子定量。这些图像是Mark Cooper提供的Zebrafish--The Living Laboratory CD中的图像系列的一部分，该CD在《细胞生物学方法》第77卷（2004年，第439-457页）中进行了描述。这是PNAS杂志1999年1月5日第96卷第157-161页的补充视频。