SGER: GFP-Based Imaging for Cryobiological Research: A Cryosurgical Application

该研究的短期目标是证明最近开发的一类荧光分子探针可以用作原位组织活力探针。具体来说，由分子生物学家开发的绿色荧光蛋白（GFP）荧光探针将与动物皮瓣窗口腔结合使用，以实现连续，动态，三维，体内组织/肿瘤观察和与细胞活力相关的荧光测量。预计这种方法的可行性的成功演示将允许令人兴奋的和新颖的后续研究，将在细胞和组织水平上提供前所未有的关于冻结/解冻损伤的细节。可以预见的是，当应用于与血管损伤相关的问题时，该方法将特别富有成效，这正是目前最先进的研究定义为最重要的问题之一。这项探索性研究的成功完成也有望使我们更好地理解我们目前知识中的另一个主要缺陷：在“慢”冷冻速率下对分离细胞以及体外和体内组织的冷冻/解冻损伤机制。最后，使用gfp和转染的免疫重要细胞的方法的未来发展可能会导致对细胞介导的冷冻免疫反应的某些方面的更好理解，这是冷冻手术和组织冷冻中的另一个重要问题。这项研究的长期目标是开发冷冻手术设备和程序，使冷冻手术成为一种成本效益高、微创的方法，可以选择性地杀死肿瘤，同时避免损伤周围的正常组织，并尽可能多地治疗癌症。目前的技术水平限制了现有冷冻手术设备和工艺的优化，以及新设备和程序的开发，这些设备和程序可以将冷冻手术的使用扩展到其他应用。限制因素是冷冻和解冻杀死癌组织的机制尚不清楚。这些机制仍然未知的部分原因与目前用于评估体内组织活力的相对粗糙的方法有关。这项研究基金将支持一名机械工程专业的女研究生，她正在为她的博士论文研究这个主题。她打算在生物医学工程领域从事学术事业。这项研究已经吸引了两名工科本科女生，她们开始参与其中，并表示有兴趣作为研究小组的一部分继续研究生学习。

9816734McGrathThe short-term objective of the research is to demonstrate that a recently developed class of fluorescent molecular probes can be used as in situ tissue viability probes. Specifically, green fluorescent protein (GFP) fluorescent probes developed by molecular biologists will be used in conjunction with an animal skin flap window chamber to allow continuous, dynamic, three-dimensional, in vivo tissue/tumor observations and measurements of fluorescence related to cellular viability. It is expected that the successful demonstration of the feasibility of this approach would allow for exciting and novel subsequent research that would provide an unprecedented level of detail regarding freeze/thaw damage at the cellular and tissue level. It is anticipated that the method will be especially productive when applied to issues related to vasculature injury, which are precisely the issues which state-of-the art research defines as one of the most important to address at this time. Successful completion of this exploratory research is also expected to lead to an improved understanding of the another major shortcoming in our current knowledge: the mechanisms of freeze/thaw injury at "slow" freezing rates for isolated cells as well as for tissues in vitro and in vivo. Finally, future developments of the method using GFPs and transfected immunologically-significant cells could lead to an improved understanding of certain aspects of cell-mediated cryoimmune responses, another important issue in cryosurgery and tissue freezing in general.The long term goal of the research is aimed at developing cryosurgical devices and procedures which would make cryosurgery a cost effective, minimally invasive method for selectively killing tumors while avoiding injury to surrounding normal tissues and to do so for as many cancers as possible. The current state of the art limits the optimization of existing cryosurgical devices and processes as well as the development of new devices and procedures that could extend the use of cryosurgery to other applications. The limiting factor is that the mechanisms by which the freezing and thawing kill the cancerous tissue is not understood well enough. Part of the reason that these mechanisms remain unknown is related to the relatively crude means currently used for assessing viability in tissues in vivo. This research grant will support a woman mechanical engineering graduate student who is conducting research on this topic for her Ph.D. thesis. She intends to pursue an academic career in the field of biomedical engineering. The research has already attracted two female undergraduate engineering students who are beginning to get involved with it and who have expressed interest to pursue graduate study as a part of the research group as a result.