PHYLOGENETIC STAINS - RIBOSOMAL RNA-BASED PROBES FOR THE IDENTIFICATION OF SINGLE CELLS
PHYLOGENETIC STAINS - RIBOSOMAL RNA-BASED PROBES FOR THE IDENTIFICATION OF SINGLE CELLS
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DOI：
10.1126/science.2466341
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发表时间：
1989-03-10
期刊：
影响因子：
56.9
通讯作者：
PACE, NR
中科院分区：
文献类型：
--
作者：
DELONG, EF;WICKHAM, GS;PACE, NR
利用一种新的染色方法实现了微生物单细胞系统发育的快速鉴定。甲醛固定的，完整的细胞与16S核糖体RNA（rRNA）互补的荧光标记的寡脱氧核苷酸杂交，并通过荧光显微镜观察。由于rRNA在细胞中的丰度，荧光探针与单个细胞的结合很容易可视化。系统发生鉴定是通过使用与系统发生组特异性16S rRNA序列互补的寡核苷酸（长度17至34个核苷酸）来实现的。适当的探针可以由区分主要界（真核生物、真细菌、古细菌）和密切相关的生物体的寡核苷酸序列组成。同时使用多个探针，用不同的荧光染料标记，允许在相同的显微镜视野中识别不同的细胞类型。定量显微荧光法表明，结合大肠杆菌的rRNA特异性探针的量随核糖体含量而变化，因此反映了生长速率。
Rapid phylogenetic identification of single microbial cells was achieved with a new staining method. Formaldehyde-fixed, intact cells were hybridized with fluorescently labeled oligodeoxynucleotides complementary to 16S ribosomal RNA (rRNA) and viewed by fluorescence microscopy. Because of the abundance of rRNA in cells, the binding of the fluorescent probes to individual cells is readily visualized. Phylogenetic identification is achieved by the use of oligonucleotides (length 17 to 34 nucleotides) that are complementary to phylogenetic group-specific 16S rRNA sequences. Appropriate probes can be composed of oligonucleotide sequences that distinguish between the primary kingdoms (eukaryotes, eubacteria, archaebacteria) and between closely related organisms. The simultaneous use of multiple probes, labeled with different fluorescent dyes, allows the identification of different cell types in the same microscopic field. Quantitative microfluorimetry shows that the amount of an rRNA-specific probe that binds to Escherichia coli varies with the ribosome content and therefore reflects growth rate.