Biopsy and Freezing of Later-stage Mouse Blastocysts Using the Dracula Pipette

描述（由申请人提供）：胚胎植入前的程序是许多生物医学研究的基石；然而，在所有情况下，程序都是非常困难和微妙的，需要非常熟练的技术专家和昂贵的设备。与这些程序相关的困难往往导致大多数此类工作被大多数生物医学研究小组所避免。当必须执行时，通常外包给专门的实验室或设施。这样做的两个缺点是：(1)由于这些程序的成本和难度，许多重要的问题可能被避免；(2)由于冷冻保存的成本和/或复杂性过高，许多独特的和有潜在价值的小鼠品系可能丢失或放弃。GeneSearch公司开发了一种新型的同轴胚胎操作工具，有望解决这些问题。这款工具的创新之处在于其简单的设计，而不是在胚胎的两极分别使用“握住”和“操纵”移液器，德古拉将操作移液器放在握住的移液器中。主要的优点是，操作移液器接触胚胎的区域是由持有移液器所传递的调节力所限制的。该工具的原始原型第一次实现了巨大的进步

DESCRIPTION (provided by applicant): Procedures on pre-implantation embryos are a cornerstone of much biomedical research; however in all cases, the procedures are very difficult and delicate, requiring exceptionally skilled technical experts and expensive equipment. The difficulties associated with these procedures often results in most such work being avoided by most biomedical research groups. When it must be performed, it is generally out-sourced to dedicated laboratories or facilities. Two downsides to this are: (1) many important questions are likely avoided due to the cost and difficulty of these procedures; and (2) many unique and potentially valuable mouse lines are likely lost or abandoned because the cost and/or complexity of cryopreservation are prohibitive. GeneSearch, Inc. has developed a novel co-axial embryo manipulation tool that promises to address these issues. The innovative elegance of this tool lies in its simple design, in which, rather than having separate "holding" and "manipulating" pipettes that work at opposite poles of the embryo, the Dracula has the manipulating pipette within the holding pipette. The major advantage is that the manipulating pipette contacts the embryo in a region that is constrained by the regulated forces imparted by the holding pipette. Original prototypes of the tool allowed, for the first time, dramatic advances in cryopreservation of large hatched embryos such as those of the llama and horse. Now, GeneSearch is scaling down the size of the tool, using new materials for some parts, and improving its handling specifically for manipulation of mouse-sized embryos. Phase I of the proposed study aims to optimize the "Mouse Dracula" tool for mouse embryological procedures including embryo cryopreservation, production of chimeric mice by ES cell injection, and survival-biopsy for genetic analyses. To accomplish this, we have proposed three specific aims, which (1) optimize tool performance, (2) develop improved protocols for mouse embryo cryopreservation, and (3) test the performance of the tool for chimera production and survival-biopsy-based genetic analyses. This is a collaborative research agreement between a Montana-based small business, GeneSearch Inc., and an embryologist/molecular geneticist at Montana State University. The goal of this Phase I STTR project is to prepare the Mouse Dracula tool to move on to Phase II market-oriented development within a one-year period.