NeuroInDx, Inc., Signal Hill, California, United States of America.
PLoS One. 2012;7(7):e41564. doi: 10.1371/journal.pone.0041564. Epub 2012 Jul 24.
We developed a novel, highly accurate, capillary based vacuum-assisted microdissection device CTAS-Cell and Tissue Acquisition System, for efficient isolation of enriched cell populations from live and freshly frozen tissues, which can be successfully used in a variety of molecular studies, including genomics and proteomics. Specific diameter of the disposable capillary unit (DCU) and precisely regulated short vacuum impulse ensure collection of the desired tissue regions and even individual cells. We demonstrated that CTAS is capable of dissecting specific regions of live and frozen mouse and rat brain tissues at the cellular resolution with high accuracy. CTAS based microdissection avoids potentially harmful physical treatment of tissues such as chemical treatment, laser irradiation, excessive heat or mechanical cell damage, thus preserving primary functions and activities of the dissected cells and tissues. High quality DNA, RNA, and protein can be isolated from CTAS-dissected samples, which are suitable for sequencing, microarray, 2D gel-based proteomic analyses, and Western blotting. We also demonstrated that CTAS can be used to isolate cells from native living tissues for subsequent recultivation of primary cultures without affecting cellular viability, making it a simple and cost-effective alternative for laser-assisted microdissection.
我们开发了一种新颖、高度精确的基于毛细管的真空辅助微切割装置 CTAS-Cell 和 Tissue Acquisition System，用于从活体和新鲜冷冻组织中高效分离富集的细胞群体，可成功应用于各种分子研究，包括基因组学和蛋白质组学。一次性毛细管单元 (DCU) 的特定直径和精确调节的短真空脉冲确保了所需组织区域甚至单个细胞的采集。我们证明了 CTAS 能够以高精确度对活体和冷冻的小鼠和大鼠脑组织的特定区域进行细胞分辨率的解剖。基于 CTAS 的微切割避免了对组织进行潜在有害的物理处理，如化学处理、激光辐射、过度加热或机械细胞损伤，从而保持了被解剖细胞和组织的主要功能和活性。从 CTAS 切割的样本中可以分离出高质量的 DNA、RNA 和蛋白质，适用于测序、微阵列、基于 2D 凝胶的蛋白质组学分析和 Western blot。我们还证明了 CTAS 可用于从天然活体组织中分离细胞，以便随后进行原代培养物的再培养，而不会影响细胞活力，使其成为激光辅助微切割的简单且具有成本效益的替代方法。
