Andy Miller, Sarah Webb (2011) CIL:12817, Danio rerio. CIL. Dataset

A single bidirectional calcium wave. The prewave pulse lasts only for 0.6 sec in this time-lapse sequence. This sequence covers 350 sec of development at 50x real time. The movie represents a 30-sec integration window moving in 5-sec steps. 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.

本数据集呈现单一的双向钙波。在该时间推移序列中，前波脉冲仅持续0.6秒。本序列涵盖了350秒的发育过程，相当于实际时间的50倍。影片中展示了30秒的积分窗口，以5秒为步长移动。受精斑马鱼卵在产卵后5分钟内收集，经过酶解去卵黄囊，并注入约0.9 nl的1%重组f-aequorin溶液，其中含有100 mM KCl、5 mM Mops和50 μM EDTA。在成像过程中，胚胎被维持在28°C的温度下，置于含有青霉素（0.5 mg/ml）、链霉素（5,000单位/ml；Sigma）和0.5%甲基纤维素30% Danieau's培养基中。成像是在Science Wares公司（Falmouth，MA）生产的Photon Imaging Microscope上进行的，该显微镜配备了带有电阻阳极输出的光子计数空间探测器（Photek，St. Albens-on-Sea，英国）。以时间标记的八位x--y坐标流（256 × 256像素）形式的数字化探测器输出被用于构建时间推移成像序列。成像系统软件允许根据选定的积分时间分析原始光子数据流，生成的图像帧保持精确的光子量化，每像素最多256个光子。这些图像是Zebrafish--The Living Laboratory CD图像系列的一部分，由Mark Cooper提供，并在《细胞生物学方法》第77卷（2004年，第439-457页）中描述。这是PNAS杂志1999年1月5日第96卷第157-161页上的补充视频。