Fluorimetric Technique for Monitoring Changes in the Level of Reduced Nicotinamide Nucleotides in Continuous Cultures of Microorganisms

Harrison, David E.; Chance, Britton

doi:10.1128/aem.19.3.446-450.1970

Cited by 116 publications

(29 citation statements)

References 8 publications

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“…The tryptophan fluorescence signal is directly related to cell mass concentration, whereas NADH fluorescence is perturbed by environmental changes. The effect of these changes on the NADH fluorescence is shown in Figures 3 and 4 and has also been studied by various researchers (Harrison and Chance, 1970;Zabriskie and Humphrey, 1978;Ristroph et al, 1977;Armiger et al, 1986).…”

Section: Discussionmentioning

confidence: 68%

“…The difficulties in measuring small absorbances in cell suspensions led other researchers (Duysens and Amesz, 1957; Chance et al, 1964;Harrison and Chance, 1970; Zabriskie and Humphrey, 1978) to investigate the use of fluorescence for measurements in cell culture. These early studies concentratad on the fluorescence from intracellular reduced nicotinamide adenine dinucleotide (NADH) or reduced nicotinamide dinucleotide phosphate (NADPH), which fluoresces around 460 nm when excited with light between 350 and 370 nm.…”

Section: Introductionmentioning

confidence: 99%

“…The rapid cycling between the oxidized and reduced states of the intracellular pyridine nucleotide was observed upon the addition of glucose or in the aerobic-anaerobic transition of yeast. Harrison and Chance (1970) used a continuous culture of Klebsiella aerogenes to evaluate NADH fluorescence for process monitoring and control. The NADH fluorescence signal was found to be sensitive to the dissolved oxygen (DO) concentration.…”

Section: Introductionmentioning

confidence: 99%

“…These instruments use a mercury lamp with suitable filters for the excitation source and detect the NADH fluorescence near 460 nm. In some studies, good correlation between cell mass and the fluorescence signal was reported (Ristroph et al, 19771, or a casual relation was established between the fluorescence signal and oxygen deprivation (Harrison and Chance, 1970). NADH fluorescence is only a good indicator of cell density when temperature, pH, and dissolved oxygen conditions are held constant and the energy substrate is in excess.…”

Section: Introductionmentioning

confidence: 99%

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In Situ Fluorescence Cell Mass Measurements of Saccharomyces cerevisiae Using Cellular Tryptophan

Horvath

Glazier

Spangler

1993

Biotechnology Progress

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This work describes a new spectroscopic optical fiber/rod technique for in situ real time measurement of cell mass and product concentrations in bioreactors using intrinsic fluorescence. The variable excitation/emission wavelength capability of this sensor allows for species-selective measurement during fermentations. Cell mass (tryptophan) and product concentrations (pyridoxine) have been measured during fermentations of Saccharomyces cerevisiae. The effects of varying substrate concentration and oxygen concentration on the observed cell mass signals are eliminated by direct measurement of cell mass, as opposed to indirect measurement schemes such as those using NADH fluorescence. The sensor is robust and able to undergo many cycles of in situ steam sterilization without degradation, and its fluorescence signal is linear with concentration for all species studied in this work. Tryptophan fluorescence from yeast is shown to be a better measure of cell mass than NADH fluorescence.

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Section: Discussionmentioning

confidence: 68%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

In Situ Fluorescence Cell Mass Measurements of Saccharomyces cerevisiae Using Cellular Tryptophan

Horvath

Glazier

Spangler

1993

Biotechnology Progress

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“…In that case increasing NADH reduction state is likely to arise partly from the 40 min period ultradian clock [7], although other causes of altered intracellular conditions cannot be excluded. Although respiratory oscillations have been described in bacteria [28], Schizosaccharomyces pombe [18,29], S. cerevisiae [7,8] and a number of protozoa [6], there have been rather few studies on the dynamics of the redox activities of mitochondria in situ in growing organisms or cells. The study of the in vivo oscillatory phenomena observed and described here are only made possible by recent developments in 2-photon laser scanning microscopy whereby simultaneous digital acquisition of intensities of fluorescence emission from different fluorophores is achieved.…”

Section: Discussionmentioning

confidence: 99%

Single and cell population respiratory oscillations in yeast: A 2‐photon scanning laser microscopy study

et al. 2006

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Two-photon scanning laser and confocal microscopies were used to image metabolic dynamics of single or cell populations of Saccharomyces cerevisiae strain 28033. Autofluorescence of reduced nicotinamide nucleotides, and mitochondrial membrane potential (DWm), were simultaneously monitored. Spontaneous, large-scale synchronized oscillations of NAD(P)H and DWm throughout the entire population of yeasts occurred under perfusion with aerated buffer in a continuous single-layered film of organisms. These oscillations stopped in the absence of perfusion and the intracellular NAD(P)H pool became reduced. Individual mitochondria within a single yeast also showed in-phase synchronous responses with the cell population, in both tetramethylrhodamine ethyl ester (or tetramethylrhodamine methyl ester) and autofluorescence. A single, localized, laser flash also triggered mitochondrial oscillations in single cells suggesting that the mitochondrion may behave as an autonomous oscillator. We conclude that spontaneous oscillations of S. cerevisiae mitochondrial redox states and DWm occur within individual yeasts, and synchrony of populations of organisms indicates the operation of an efficient system of cell-cell interaction to produce concerted metabolic multicellular behaviour on the minute time scale in both cases.

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Fluorescence Techniques for Bioprocess Monitoring

Anton

Lindemann

Hitzmann

et al. 2010

Encyclopedia of Industrial Biotechnology

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Modern bioprocess control requires instantaneous data acquisition and in‐time evaluation of bioprocess variables. Optical sensor systems are of increasing interest in bioprocess monitoring because they are very sensitive and offer the possibility of on‐line, in situ , and noninvasive process monitoring. In this regard, on‐line fluorescence spectroscopy is a useful tool in the development of methods for bioprocess monitoring because many cellular substances and media components are biogenic fluorophors. This technique and the application of chemometric methods offer a deeper insight into biological processes and thus process monitoring, optimization, and control is accessible. In this review, an overview of the developments and applications of on‐line fluorescence techniques for bioprocess monitoring over the last 25 years is given with emphasis on 2D fluorescence spectroscopy.

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Fluorimetric Technique for Monitoring Changes in the Level of Reduced Nicotinamide Nucleotides in Continuous Cultures of Microorganisms

Cited by 116 publications

References 8 publications

In Situ Fluorescence Cell Mass Measurements of Saccharomyces cerevisiae Using Cellular Tryptophan

In Situ Fluorescence Cell Mass Measurements of Saccharomyces cerevisiae Using Cellular Tryptophan

Single and cell population respiratory oscillations in yeast: A 2‐photon scanning laser microscopy study

Fluorescence Techniques for Bioprocess Monitoring

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