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

9816734McGrath这项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性探针。具体地说，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室结合使用，以实现连续、动态、三维、活体组织/肿瘤观察和与细胞活性相关的荧光测量。预期这一方法的可行性的成功证明将使随后的研究变得令人兴奋和新颖，这些研究将在细胞和组织层面上提供关于冻融损害的前所未有的详细程度。预计当应用于与血管损伤相关的问题时，该方法将特别有成效，这正是最先进的研究定义为目前最重要的问题之一。这项探索性研究的成功完成还有望提高对我们目前知识中另一个主要缺陷的理解：对分离的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和转基因的免疫显著细胞的方法的未来发展可能有助于更好地理解细胞介导的冷冻免疫反应的某些方面，这是冷冻外科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外科设备和程序，使冷冻外科成为一种具有成本效益的微创方法，在选择性杀死肿瘤的同时避免对周围正常组织的伤害，并尽可能多地治疗癌症。目前的技术水平限制了现有冷冻外科设备和工艺的优化以及新设备和工艺的开发，这些新设备和工艺可以将低温外科的使用扩展到其他应用。限制因素是，冷冻和解冻杀死癌症组织的机制还不够清楚。这些机制仍不清楚的部分原因与目前用于评估体内组织活性的相对粗糙的方法有关。这笔研究经费将用于资助一名机械工程专业的女研究生，她正在为自己的博士论文进行这一课题的研究。她打算在生物医学工程领域追求学术生涯。这项研究已经吸引了两名工程专业的女本科生，她们开始参与这项研究，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.