Fluorescence microscopy of NaCl-stressed, elongated Salmonella and Listeria cells reveals the presence of septa in filaments
Fluorescence microscopy of NaCl-stressed, elongated Salmonella and Listeria cells reveals the presence of septa in filaments
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DOI：
10.1016/j.ijfoodmicro.2006.04.026
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发表时间：
2006-12-01
影响因子：
5.4
通讯作者：
Beumer, Rijkelt R.
中科院分区：
文献类型：
--
作者：
Hazeleger, Wilma C.;Dalvoorde, Marinka;Beumer, Rijkelt R.
检查施加应激后肠炎沙门氏菌和单核细胞增生李斯特氏菌的细胞形态。通过将细菌铺在含有 5-10% NaCl 的胰蛋白胨大豆琼脂 (TSA) 板上，对细胞进行应激。随后将板在 25°C 下孵育 6 天。最后，收获细胞并进行不同的荧光探针处理，以观察细长细胞中可能存在的隔膜。使用染色剂 4',6-二脒基-2-苯吲哚 (DAPI)（一种优先染色双链 DNA 的蓝色荧光核酸染色剂）可以清楚地显示沙门氏菌和李斯特菌细胞中存在黑点，可能是不存在核酸的细胞分区。另一种染色剂 FM 4-64（一种用于内膜红色染色的亲脂性苯乙烯基染料）显示李斯特菌细胞中存在强荧光斑点，可能表明细胞膜的存在。然而，对于沙门氏菌，FM 4-64 未能成功揭示细丝中的隔膜。对伸长的李斯特菌细胞进行双重染色，显示 DAPI 染色（富含 DNA 的斑点）中具有高荧光的区域，而 FM 4-64 染色（膜斑点）中含有低荧光的区域，反之亦然，这证实了伸长的细胞确实由几个正常大小的细胞组成。 (c) 2006 Elsevier B.V. 保留所有权利。
The cell morphology of Salmonella enteritidis and Listeria monocytogenes after the application of stress was examined. Cells were stressed by plating the bacteria on Tryptone Soya Agar (TSA) plates, with 5-10% NaCl. The plates were subsequently incubated for 6 days at 25 degrees C. Finally, the cells were harvested and subjected to different fluorescent probes in order to visualize the possible presence of septa in elongated cells. Use of the stain 4',6-Diamidino-2-phenytindole (DAPI), which is a blue fluorescent nucleic acid stain that preferentially stains double-stranded DNA, showed clearly the presence of dark spots, probably cellular partitions where no nucleic acids were present, in both Salmonella and Listeria cells. Another stain, FM 4-64, a lipophilic styryl dye for red staining of the inner membrane, showed the presence of highly fluorescent spots in Listeria cells, probably indicating the presence of membranes. For Salmonella, however, FM 4-64 was not successful in revealing septa in filaments. Double staining applied to elongated Listeria cells showed areas with high fluorescence in DAPI-staining (DNA-rich spots) which contained low fluorescence in FM 4-64-staining (membrane spots) and vice versa, which is a confirmation that the elongated cells are indeed composed of several normal size cells. (c) 2006 Elsevier B.V. All rights reserved.