caa a22 4500 477105785 CHVBK 20180405111553.0 cr unu---uuuuu 170330e19960101xx s 000 0 eng 10.1007/BF01569921 doi (NATIONALLICENCE)springer-10.1007/BF01569921 A dual fluorescence technique for visualization of Staphylococcus epidermidis biofilm using scanning confocal laser microscopy [Elektronische Daten] [B Sanford, A de Feijter, M Wade, V Thomas] A new dual fluorescence technique is described which, when combined with scanning confocal laser microscopy (SCLM), can be used to visualize the components of biofilm produced byStaphylococcus epidermidis. Chemostat cultures of RP62A (a well-characterized slime-producing strain ofS. epidermidis) were used to produce mature biofilm on polyvinylcholoride (PVC) disks immobilized in a modified Robbins device using a ‘seed' and ‘feed' model system. Serial horizontal and vertical optical thin sections, as well as three-dimensional computer reconstructions, were obtained onin situ biofilm using the dual fluorescence procedure. Bacteria were visualized by green autofluorescence excited at 488 nm with an Argon laser. Cell-associated and exocellular matrix material (slime) was visualized by red fluorescence excited at 568 nm with a Krypton laser after interaction of the biofilm with Texas Red-labeled wheat germ agglutinin which is a slime-specific lectin marker. Structural analysis revealed that the cocci grew in slime-embedded cell clusters forming distinct conical-shaped microcolonies. Interspersed open channels served to connect the bulk liquid with the deepest layers of the mature, hydrated biofilm which increased overall surface area and likely facilitated the exchange of nutrients and waste products throughout the biofilm. The combined dual fluorescence technique and SCLM is potentially useful as a specific noninvasive tool for studying the effect of antimicrobial agents on the process of biofilm formation and for the characterization of the architecture ofS. epidermidis biofilm formedin vivo andin vitro on medical grade virgin or modified inert polymer surfaces. Society for Industrial Microbiology, 1996 Staphylococcus epidermidis nationallicence biofilm nationallicence laser scanning confocal microscopy nationallicence slime nationallicence lectin marker nationallicence Sanford B. Department of Microbiology, The University of Texas Health Science Center at San Antonio, Floyd Curl, 78284, San Antonio, Texas, USA aut de Feijter A. Meridian Instruments, Inc, Floyd Curl, 48864, Okemos, Michigan, USA aut Wade M. Meridian Instruments, Inc, Floyd Curl, 48864, Okemos, Michigan, USA aut Thomas V. Department of Microbiology, The University of Texas Health Science Center at San Antonio, Floyd Curl, 78284, San Antonio, Texas, USA aut Journal of Industrial Microbiology Springer-Verlag 16/1(1996-01-01), 48-56 0169-4146 16:1<48 1996 16 10295 https://doi.org/10.1007/BF01569921 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01569921 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sanford B. Department of Microbiology, The University of Texas Health Science Center at San Antonio, Floyd Curl, 78284, San Antonio, Texas, USA aut NATIONALLICENCE 700 1- de Feijter A. Meridian Instruments, Inc, Floyd Curl, 48864, Okemos, Michigan, USA aut NATIONALLICENCE 700 1- Wade M. Meridian Instruments, Inc, Floyd Curl, 48864, Okemos, Michigan, USA aut NATIONALLICENCE 700 1- Thomas V. Department of Microbiology, The University of Texas Health Science Center at San Antonio, Floyd Curl, 78284, San Antonio, Texas, USA aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology Springer-Verlag 16/1(1996-01-01), 48-56 0169-4146 16:1<48 1996 16 10295 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer