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STED nanoscopy of KK114-stained pathogenic bacteria.
Journal of Biophotonics ( IF 2.0 ) Pub Date : 2020-06-23 , DOI: 10.1002/jbio.202000097 Massimiliano Lucidi 1 , Radu Hristu 2 , Lorenzo Nichele 1 , George A Stanciu 2 , Denis E Tranca 2 , Alina Maria Holban 3 , Paolo Visca 4 , Stefan G Stanciu 2 , Gabriella Cincotti 1
Journal of Biophotonics ( IF 2.0 ) Pub Date : 2020-06-23 , DOI: 10.1002/jbio.202000097 Massimiliano Lucidi 1 , Radu Hristu 2 , Lorenzo Nichele 1 , George A Stanciu 2 , Denis E Tranca 2 , Alina Maria Holban 3 , Paolo Visca 4 , Stefan G Stanciu 2 , Gabriella Cincotti 1
Affiliation
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Super‐resolution microscopy techniques can provide answers to still pending questions on prokaryotic organisms but are yet to be used at their full potential for this purpose. To address this, we evaluate the ability of the rhodamine‐like KK114 dye to label various types of bacteria, to enable imaging of fine structural details with stimulated emission depletion microscopy (STED). We assessed fluorescent labeling with KK114 for eleven Gram‐positive and Gram‐negative bacterial species and observed that this contrast agent binds to their cell membranes. Significant differences in the labeling outputs were noticed across the tested bacterial species, but importantly, KK114‐staining allowed the observation of subtle nanometric cell details in some cases. For example, a helix pattern resembling a cytoskeleton arrangement was detected in Bacillus subtilis. Furthermore, we found that KK114 easily penetrates the membrane of bacterial microorganism that lost their viability, which can be useful to discriminate between living and dead cells.
中文翻译：
KK114染色的病原菌的STED纳米显微镜检查。
超分辨率显微镜技术可以为原核生物悬而未决的问题提供答案，但尚未充分发挥其潜力。为了解决这个问题，我们评估了若丹明样KK114染料标记各种类型细菌的能力，从而能够使用受激发射损耗显微镜（STED）对精细的结构细节进行成像。我们评估了KK114对11种革兰氏阳性和革兰氏阴性细菌种类的荧光标记，并观察到该造影剂与细胞膜结合。在所测试的细菌物种之间，标记输出存在显着差异，但重要的是，KK114染色可在某些情况下观察细微的纳米细胞细节。例如，在细胞中检测到类似于细胞骨架排列的螺旋模式枯草芽孢杆菌。此外，我们发现KK114容易穿透丧失生存能力的细菌微生物膜，这对于区分活细胞和死细胞很有用。
更新日期：2020-06-23
中文翻译：
KK114染色的病原菌的STED纳米显微镜检查。
超分辨率显微镜技术可以为原核生物悬而未决的问题提供答案，但尚未充分发挥其潜力。为了解决这个问题，我们评估了若丹明样KK114染料标记各种类型细菌的能力，从而能够使用受激发射损耗显微镜（STED）对精细的结构细节进行成像。我们评估了KK114对11种革兰氏阳性和革兰氏阴性细菌种类的荧光标记，并观察到该造影剂与细胞膜结合。在所测试的细菌物种之间，标记输出存在显着差异，但重要的是，KK114染色可在某些情况下观察细微的纳米细胞细节。例如，在细胞中检测到类似于细胞骨架排列的螺旋模式枯草芽孢杆菌。此外，我们发现KK114容易穿透丧失生存能力的细菌微生物膜，这对于区分活细胞和死细胞很有用。
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