Advanced cell sorting technology to properly investigate biological mechanisms in the presence of cellular heterogeneity

在研究控制细胞对微环境的反应的生物学机制时，通常采取一种或多种单独的测量来表征给定的反应。这些措施有效地呈现了被研究人群中每个细胞的平均值。出于这个原因，至关重要的是，被调查的细胞群体是同质的；换句话说，每个细胞彼此之间应该非常相似，它们对刺激的反应也应该相似。否则，所得的测量结果可能不能准确地代表所研究的现象，除非这些异质细胞彼此分离。在细胞微环境或其他形式的外源刺激驱使细胞分化成一种或多种新的、高度特化的细胞类型的情况下，异质性是预期的。在这里，根据细胞的分化表型分离或分类细胞的需要是至关重要的，因为它将允许研究人员使用他们强大的细胞和分子技术工具包和措施来了解这些分化亚群的生物学基础。显然，为了测试细胞和分子生物学中新颖和创新的假设，先进的仪器将异质细胞类型物理分类并分离到定义的群体中是必不可少的。不幸的是，对于我们的研究团队来说，代表nserc资助的来自劳伦森大学、北安大略医学院和健康科学北方研究所的研究人员，在400公里半径内没有足够的手段来实现这一目标。有了这笔资金，我们的目标是购买一台自动细胞分选仪器，这将使我们能够从混合群体中分离出两种表型不同的细胞类型，然后我们可以使用传统的细胞和分子生物学技术进行分析。这将极大地推进我们的研究目标，使我们的研究人员能够研究他们目前无法获得的独特细胞亚群的信号机制；使用尖端的基于crispr的筛选工具获得罕见的基因突变细胞，可能揭示肿瘤细胞转化的本质；并从一个看似同质的群体中挑选出一些细胞，这些细胞能够抵抗辐射的致癌效应，原因尚不清楚。此外，将这些先进的仪器添加到我们目前的细胞生物学工具综合系统中，将丰富我们为安大略省东北部的学生和其他学员提供的高质量培训环境。这将提高他们的技能的市场竞争力，同时为区域自然科学、生物技术和医疗行业提供更好的候选人来推动其业务的创新。

In studies of the biological mechanisms governing cellular responses to their microenvironment it is common to take one or more individual measurements to characterize a given response. These measures effectively present the average of every cell within that population being studied. For this reason, it is critical that the cellular population being investigated is homogenous; in other words, every cell should be very similar to one another and their responses to a stimulus should be similar as well. If not, the resultant measurement may not accurately represent the phenomenon being studied unless those heterogenous cells are separated from one another. In the case where a cell's microenvironment or some other form of exogenous stimuli drives that cell to differentiate into one or more new, highly specialized cell types, heterogeneity is expected. Here, the need to separate or sort cells according to their differentiated phenotype is of utmost importance, as it will allow investigators to use their powerful toolkit of cellular and molecular techniques and measures to understand the biological underpinnings of those differentiated subsets. Clearly, to test novel and innovative hypotheses in cellular and molecular biology, advanced instrumentation to physically sort and separate heterogenous cell types into defined populations is fundamental. Unfortunately, for our research team representing NSERC-funded investigators from Laurentian University, the Northern Ontario School of Medicine University and Health Sciences North Research Institute, there are no adequate means to achieve this goal within a 400 km radius. With this funding, we aim to purchase an automated cell sorting instrument that will allow us to isolate up to two phenotypically distinct types of cells from a mixed population, which we can then analyze using traditional cellular and molecular biology techniques. This will greatly advance our research goals, allowing our investigators to study signalling mechanisms of unique cell subsets that they are currently unable to obtain; use cutting edge CRISPR-based screening tools to obtain rare genetic mutant cells that may uncover the nature of neoplastic cell transformation; and sort out cells within a seemingly homogenous population that for some yet to be discovered reason are able to resist the cancer causing effects of radiation. Furthermore, adding such advanced instrumentation to our current comprehensive system of cell biology tools will enrich the highly-qualified training environment that we provide to students and other trainees in Northeastern Ontario. This will improve the marketability of their skills, while providing regional natural sciences, biotechnology and medical industries better candidates to drive innovation in their businesses.