分离鉴别未知微生物物种成为可能
世界上存在着数以百万种的微生物种,但是截至目前科学家们仅仅可以鉴别出这其中的很少一部分,许多种微生物由于不能在实验室条件下生存所以目前研究人员并不能对其进行鉴定;近日来自美国东北大学等处的研究人员,工程学家Edgar Goluch和生物学家Slava Epstein通过研究开发了一种新型设备,其可以对环境中的任何一种单一微生物种进行培养,进而进行后续研究和鉴定,相关研究成果于7月1日刊登于PLOS ONE上。
此前研究中Goluch利用掺入渗透膜的设备就可以将隐藏在环境中的微生物暴露于营养物质中进而对其研究,但是在自然环境中物种之间存在激烈的竞争,所以长期以来研究者们可以分离得到的微生物种依然很少。
而这项研究中,他们两人解决了这个问题,这种新型设备可以使得单一的细菌细胞通过通道进入到包含有细菌“食物”的小室中,而细菌细胞进入的显微通道,其仅比细菌细胞稍窄一些。这种显微通道如此狭窄以至于当进入一个细菌细胞后,就可以阻断其余细胞的进入;在小室中细菌的细胞就可以开始增殖,当其增殖到纯种样本并且充满整个箱体后,研究者就会对其进行收集并且进行鉴定。
研究者表示,这种新型设备可以对不同实验室条件下的不同细胞类型进行分离,在第一组实验中研究者分离了细胞大小略有不同的大肠杆菌和铜绿假单胞菌,在第二组实验中他们分离了细胞大小相似而细胞形状不同的玫瑰杆菌属(Roseobacter)和Pscyhoserpens菌属;运用红光或者绿光的不同标记,最终研究者利用该设备分离了不同种类的细胞。最终的实验结果证实了研究者的假设,即生长在食物暗箱中的细菌细胞就是当时进入到暗箱狭窄通道细胞的后代细胞。相关研究由美国国家自然科学家基金等处提供资助。(来源:生物谷Bioon.com)
Isolation of Microorganisms Using Sub-Micrometer Constrictions
Abstract We present an automated method for isolating pure bacterial cultures from samples containing multiple species that exploits the cell\&\#39\;s own physiology to perform the separation. Cells compete to reach a chamber containing nutrients via a constriction whose cross-sectional area only permits a single cell to enter, thereby blocking the opening and preventing other cells from entering. The winning cell divides across the constriction and its progeny populate the chamber. The devices are passive and require no user interaction to perform their function. Device fabrication begins with the creation of a master mold that contains the desired constriction and chamber features. Replica molding is used to create patterned polymer chips from the master, which are bonded to glass microscope cover slips to create the constrictions. We tested constriction geometries ranging from 500 nanometers to 5 micrometers in width, 600 to 950 nanometers in height, and 10 to 40 micrometers in length. The devices were used to successfully isolate a pure Pseudomonas aeruginosa culture from a mixture that also contained Escherichia coli. We demonstrated that individual strains of the same species can be separated out from mixtures using red and green fluorescently-labeled E. coli. We also used the devices to isolate individual environmental species. Roseobacter sp. was separated from another marine species, Psychroserpens sp.
原文链接:http://www.plosone.org/article/fetchObject.action?uri=info%3
Adoi%2F10.1371%2Fjournal.pone.0101429&representation=PDF