TETRANITRO-BLUE TETRAZOLIUM REDUCTION IN <i>PROTEUS VULGARIS </i>

Leene, W.; Iterson, Woutera van

doi:10.1083/jcb.27.1.241

Cited by 13 publications

(5 citation statements)

References 6 publications

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“…Electron microscopic studies (Sedar and Burde 1965 ; Leene and Van Iterson 1965) indicate that T C reduction occurs at the inner surface of the cytoplasmic membrane and our light microscopic observations were consistent with this interpretation. We, therefore, favour the view that extracellular deposits with M T T and I N T result from diffusion of formazans through the cell envelope and subsequent re-precipitation within the mountant.…”

Section: Patterns Of Formazan Depositionsupporting

confidence: 88%

“…Many microbiologists have noted the value of tetrazolium salts as indicators of viability (Postgate 1967;Bartlett et al 1976;Urban and Jarstrand 1979;Zimmerman et al 1978;Tabor and Neihof 1982;Roszaak and Colwell 1987). These reagents have also been used for specific staining (Sedar and Burde 1965; Leene and van Iterson 1965) and biochemical reactions (Barer et al 1990;Cesari et al 1969), and as components of culture media (MacFaddin 1982;Munson 1974;Slanetz and Bartley 1957). In spite of this widespread use, microbiological studies have largely ignored the substantial body of information on the in situ chemistry of tetrazolium reactions (Altman 1976;Horobin 1982;Seidler 1991) which has underpinned their application in the demonstration and quantitation of specific enzyme activities at explicit tissue locations (Lojda et al 1979;Rost 1980;Van Noorden and Jonges 1987).…”

Section: Introductionmentioning

confidence: 99%

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Factors affecting the selection and use of tetrazolium salts as cytochemical indicators of microbial viability and activity

Thom

Horobin

Seidler³

et al. 1993

Journal of Applied Bacteriology

119

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The reduction of four tetrazolium cations (TCs), nitroblue tetrazolium (NBT), neotetrazolium (NT), methylthiazolyldiphenyltetrazolium (MTT) and iodonitrophenyltetrazolium (INT), by viable micro-organisms, immobilized on glass cover slips, was examined by light microscopy with a view to determining a systematic basis for applying these reagents as cytochemical indicators of microbial viability and activity. The potential value of histochemical information about TC reactions for developing their microbiological applications was also assessed. INT and MTT detected viable cells more readily than NBT and NT. In order to obtain cell-localized formazan, MTT required cobalt ions in the reaction mixture and INT reactions had to be assessed soon after mounting. In general, formazan deposition could be accelerated by the addition of glucose and an intermediate electron carrier (IEC) to the reaction mixture, although inhibitory effects of IECs were also detected. Cultures in exponential phase, in stationary phase and inhibited by chloramphenicol could be differentiated with MTT but not with INT. For some organisms, notably Candida albicans, Pseudomonas aeruginosa and Enterococcus faecalis. TC reactions proved to be a relatively insensitive means of demonstrating viability. Two parameters used in selecting TCs for histochemical reactions, lipophilicity and reducibility appeared to be predictive for the relative sensitivity of these reagents as indicators of cell viability. The concepts of substantivity, a measure of non-specific interactions between reagents and staining substrates, and TC oxygen sensitivity, the effect of competition between oxygen and TCs for electrons, were found to be relevant to formazan deposition in live microbes. These findings support the use of TCs as cytochemical probes of microbial activity in defined settings and the use of histochemical knowledge to support further development of these techniques.

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Section: Patterns Of Formazan Depositionsupporting

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Factors affecting the selection and use of tetrazolium salts as cytochemical indicators of microbial viability and activity

Thom

Horobin

Seidler³

et al. 1993

Journal of Applied Bacteriology

119

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“…It is interesting to note that the chondrioids adhere so firmly to the plasma membrane ( Figs. 14, 15) ; they were earlier described as being contiguous with it (17,18,24). O n the other hand, the basal granules appear to be quite separate from the membrane.…”

Section: Discussionmentioning

confidence: 94%

“…footnote, reference 20). The chondrioids themselves can readily be made visible by incubating the bacteria with either potassium tellurite (17) or tetranitro-blue tetrazolium (23,24). Reduced products of these compounds are deposited in rather restricted cytoplasmic areas contiguous with the plasma membrane.…”

Section: Introductionmentioning

confidence: 99%

BASAL BODIES OF BACTERIAL FLAGELLA IN PROTEUS MIRABILIS

Hoeniger

Iterson

Zanten

1966

The Journal of Cell Biology

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This paper investigates further the question of whether the flagella of Proteus mirabilis emerge from basal bodies. The bacteria were grown to the stage of swarmer differentiation, treated lightly with penicillin, and then shocked osmotically. As a result of this treatment, much of the cytoplasmic content and also part of the plasma membrane were removed from the cells. When such fragmented organisms were stained negatively with potassium phosphotungstate, the flagella were found to be anchored--often by means of a hook--in rounded structures approximately 50 m/z wide, thus confirming Part I of our study. In these rounded structures a more brilliant dot was occasionally observed, which we interpret as being part of the basal granule. A prerequisite for the demonstration of the basal granules within the cells was, however, the removal of both the cytoplasm and the plasma membrane from their vicinity. In some experiments, the chondrioids were "stained" positively by the incorporation into them of the reduced product of potassium tellurite. The chondrioids were here observed to be more or less circular areas from which rodlike structures extended. The chondrioids adhered so firmly to the plasma membrane that they were carried away with it during its displacement by osmotic shocking, while the basal bodies were left behind. This observation disproves our previous suggestion that the flagella might terminate in the chondrioids. The basal bodies often occur in pairs, which suggest that they could be selfreproducing particles.

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“…are responsible for substrate-dependent ion uptake, as could be demonstrated cytochemically for Proteus by VAN ITERSON & LEENE (1964) and LEENE &VAN ITERSON (1965). Since these fragments are precipitated upon treatment with organic solvents or salt (ASANO &…”

mentioning

confidence: 87%

The Distribution and Some Properties of Accumulated Silicate in Cell-Free Bacterial Extracts

Heinen

1968

Acta Botanica Neerlandica

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On incubation with silicate and a suitable substrate, cellfree extracts of Proteus mirabilis incorporate the anion Si, which is successively destributed and linked to various compounds. The distribution pattern changes with increasing incubation time: On fractionation with alcohol-ether (AES), most of the Si appears first in the alcohol-ether-insoluble (AEI) fraction reaching a maximum after 2 hours, whereafter a steady decline of the Si-concentration is observed. This fraction was found to contain water-extractable Si-compounds. The amount of Si that can be extracted from the precipitated material increases with time and temperature; maximum yield is obtained extracting the residue at 50 DC for 3 hours. The amount of waterextractable Si-compounds present in the AEI-fraction also varies with increasing incubation time: The high concentration found after 30 min rapidly declines to a minimum after 4 hours, followed by a slow increase during further incubation.Both the AEI-and AES-fraction contain acid-labile Si-compounds, liberating free silicic acid on acid hydrolysis. The percentage of these acid-labile compounds in both fractions was also found to vary with the time of incubation.

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TETRANITRO-BLUE TETRAZOLIUM REDUCTION IN PROTEUS VULGARIS

Cited by 13 publications

References 6 publications

Factors affecting the selection and use of tetrazolium salts as cytochemical indicators of microbial viability and activity

Factors affecting the selection and use of tetrazolium salts as cytochemical indicators of microbial viability and activity

BASAL BODIES OF BACTERIAL FLAGELLA IN PROTEUS MIRABILIS

The Distribution and Some Properties of Accumulated Silicate in Cell-Free Bacterial Extracts

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