Kipanga Purity N, Luyten Walter
Faculty of Pharmaceutical Sciences, KU Leuven, University of Leuven, Belgium; Department of Biology, Animal Physiology and Neurobiology Section, KU Leuven, University of Leuven, Belgium.
Department of Biology, Animal Physiology and Neurobiology Section, KU Leuven, University of Leuven, Belgium.
J Microbiol Methods. 2017 Aug;139:8-11. doi: 10.1016/j.mimet.2017.04.006. Epub 2017 Apr 21.
Adhesion is a crucial initial step in microbial biofilm formation. Firm attachment to a target surface subsequently ensures successful colonization and survival despite turbulent conditions. In the laboratory, polystyrene plates are commonly used in biofilm experiments and the 'washing/rinse steps' before staining are critical for assaying biofilm viability. However, these rinse steps risk the removal (partially or entirely) of the formed biofilm, resulting in inconsistent results. The aim of the present study was to optimize conditions for firmer biofilms, less prone to disruption and thus significantly reducing well-to-well variability. Candida albicans SC5314 was used in five different polystyrene 96-well plates from four different manufacturers. Irrespective of how gently we performed the rinse, biofilms came off certain polystyrene plates more easily compared to others. Importantly, preconditioning the polystyrene surfaces with foetal bovine serum (FBS) had a negative impact on firm biofilm attachment. Costar® plates provided the most suitable surface for firm biofilm attachment, both in the presence and absence of FBS. Substratum properties even among seemingly identical synthetic materials may influence biofilm attachment and its subsequent sturdiness, affecting experimental results.
黏附是微生物生物膜形成过程中的关键起始步骤。随后，牢固地附着在目标表面可确保在动荡环境下成功定殖和存活。在实验室中，聚苯乙烯平板常用于生物膜实验，染色前的“洗涤/冲洗步骤”对于检测生物膜活力至关重要。然而，这些冲洗步骤存在（部分或完全）去除已形成生物膜的风险，从而导致结果不一致。本研究的目的是优化条件，以形成更牢固的生物膜，使其更不易被破坏，从而显著降低孔间差异。白色念珠菌SC5314被用于来自四个不同制造商的五种不同的聚苯乙烯96孔板中。无论我们冲洗得多么轻柔，与其他板相比，某些聚苯乙烯板上的生物膜更容易脱落。重要的是，用胎牛血清（FBS）预处理聚苯乙烯表面对生物膜的牢固附着有负面影响。无论有无FBS，康宁（Costar®）板都为生物膜的牢固附着提供了最合适的表面。即使在看似相同的合成材料中，基质特性也可能影响生物膜的附着及其后续的坚固性，进而影响实验结果。
