Assessment of the effects of gramicidin, formaldehyde, and surfactants onEscherichia coli by flow cytometry using nucleic acid and membrane potential dyes

Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities

“…Crissman et al, 1978;Steen et al, 1982Steen et al, , 1986) and other stress-producing substances (e.g. Comas and Vives-Rego, 1997), and characterizing the starvation-survival response of selected bacterial species, usually those with pathogenic relevance (Thorsen et al, 1992;Lebaron and Joux, 1994). The development of fluorescent probes that indicate various aspects of cell metabolism has further stimulated this area of research (McFeters et al, 1995;Porter et al, 1996;Davey et al, 1999).…”

“…GASOL and P.A. DEL GIORGIO Mason et al 1995, Deere et al 1995 (3)) defficient Jepras et al 1995, López-Amorós et al 1995aComas and Vives-rego 1997, Beck and Huber 1997Activity of the enzime 494 / 518 cultures Nir et al 1990, Miao et al 1993 ß-galactosidase Fluorescein diacetate (FDA) cleaved by intracellular enzims 492 / 517 cultures Diaper et al 1992 CFDA, Chemchrome B, cleaved by intracellular enzims (488-520/560) cultures Diaper and Edwards 1994a, Jepras et al 1995, calcein blue AM, SFDA... Jacobsen et al 1997, Beck and Huber 1997compost Diaper and Edwards 1994bfreshwater Porter et al 1995a indicator of respiratory-chain 480 / >585 cultures Kaprelyants and Kell 1993a, López-activity Amorós et al 1997freshwater del Giorgio et al 1997b, Yamaguchi and Nasu 1997marine López-Amorós et al 1998, Sieracki et al 1999 accumulated in live cells 510 / 580 cultures Diaper et al 1992, Davey et al 1993, Kaprelyants and Kell 1992 Joux et al 1997, Jacobsen et al 1997 cells that had temporarily lost this potential and were in a state of arrested growth ("dormant" or "inactive") (Roszak and Colwell, 1987). Hutter and Eipel (1978) used Erythrosine B to label damaged yeast cells and since then many more viability probes have been put in use for bacteria (Table 3).…”

“…dyes, which are negative-charged dyes sensitive to membrane potential and preferentially stain cells with unpolarized membranes. Complementary to Rh123, they are nontoxic and do not require EDTA treatment to be used instead of PI (López-Amorós et al, 1995aDeere et al, 1995;Mason et al, 1995;Comas and Vives-Rego, 1997;Nebe-von Caron et al, 1998). Porter et al (1995a) tested some viability dyes derivatives from the FDA (CFDA, ChemChrome B, etc.)…”

Applications of Flow Cytometry to Clinical Microbiology

“…Rhodamine 123 and oxonol [bis(1,3-dibutylbarbituric acid) trimethine oxonol] are other membrane potential probes used to assess drug effects by FCM (59,150,310). Their accumulation inside bacterial cells depends on the difference in charge between the two sides of the plasma membrane.…”

“…Dye concentrations and staining times can also affect susceptibility testing by FCM (59,108,201). For example, the labeling properties of oxonol in E. coli are related to dye concentrations (201).…”