http://dbkgroup.org/Papers/davey_kell_micrev96.pdf

Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses.

A comparative analysis of the properties of two optical biosensor platforms: (1) the propagating surface plasmon resonance (SPR) sensor based on a planar, thin film gold surface and (2) the localized surface plasmon resonance (LSPR) sensor based on surface confined Ag nanoparticles fabricated by nanosphere lithography (NSL) are presented The binding of Concanavalin A (ConA) to mannose-functionalized self-assembled monolayers (SAMs) was chosen to highlight the similarities and differences between the responses of the real-time angle shift SPR and wavelength shift LSPR biosensors During the association phase in the real-time binding studies, both SPR and LSPR sensors exhibited qualitatively similar signal vs time curves However, in the dissociation phase, the SPR sensor showed an approximately 5 times greater loss of signal than the LSPR sensor A comprehensive set of nonspecific binding studies demonstrated that this signal difference was not the consequence of greater nonspecific binding to the LSPR sensor but rather a systematic function of the Ag nanoparticle's nanoscale structure Ag nanoparticles with larger aspect ratios showed larger dissociation phase responses than those with smaller aspect ratios A theoretical analysis based on finite element electrodynamics demonstrates that this results from the characteristic decay length of the electromagnetic fields surrounding Ag nanoparticles being of comparable dimensions to the ConA molecules Finally, an elementary (2 x 1) multiplexed version of an LSPR carbohydrate sensing chip to probe the simultaneous binding of ConA to mannose and galactose-functionalized SAMs has been demonstrated

A comparative analysis of localized and propagating surface plasmon resonance sensors: the binding of concanavalin a to a monosaccharide functionalized self-assembled monolayer.

Towards advanced chemical and biological nanosensors-An overview.

The still poorly explored world of microbial functioning is about to be uncovered by a combined application of old and new technologies. Bacteria, especially, are still in the dark with respect to their phylogenetic affiliations as well as their metabolic capabilities and functions. However, with the advent of sophisticated flow cytometric and cell sorting technologies in microbiological labs, there is now the possibility to gain this knowledge at the single-cell level without cumbersome cultivation approaches. Cytometry also facilitates the understanding of physiological diversity in seemingly likewise acting populations. Both individuality and diversity lead to the complex and concerted actions of microbial consortia. This review provides an overview of the state of the art in the field. It deals with the handling of microorganisms from the very beginning (i.e. sampling, and detachment and fixation procedures) and goes on to discuss the pitfalls and problems in analysing cells without any further treatment. If information cannot be gained by specific staining procedures, phylogenetic technologies, transcriptomic and proteomic approaches may be options for achieving advanced insights. All in all, flow cytometry will be a mediator technology to gain a deeper insight into the heterogeneity of populations and the functioning of microbial communities.

Functional single-cell analyses: flow cytometry and cell sorting of microbial populations and communities.

Publisher Summary Carbohydrate oxidases are found in all kingdoms of life but are mostly found in fungi. Their natural role is not always clear. Usage of molecular oxygen as electron acceptor is not a logical choice when the enzyme is part of a catabolic pathway. This chapter provides an overview of the occurrence and properties of carbohydrate oxidases. The physiological role of the different enzymes is discussed in relation to their origin, and the catalytic and structural properties are discussed in relation to their family background. It also provides a summary of the biocatalytic applications of carbohydrate oxidases. Carbohydrate oxidases are valuable enzymes for several applications. They are relatively stable and do not need expensive coenzymes. Carbohydrate oxidases are widely used in diagnostic applications, in the food and drinks industry, and for carbohydrate synthesis. They are also used for bleaching (production of H2O2) and as oxygen scavenger.

Occurrence and biocatalytic potential of carbohydrate oxidases.

Three different examples of enzymatic bioconversion in liquid membrane reactors

The development of a simple and very easy-to-use fiber optical oxygen multisensor measuring system is described. This fiber optical chemosensor (optode) is based on the fluorescence quenching of a special fluorophor by oxygen. Standard applications of the well-established amperometric oxygen sensor can be carried out with this optical sensor. Beyond that, many special applications for oxygen measurement become possible (O.S. Wolfbeis, Fiber Optic Chemical Sensors and Biosensors, vols. I and II, CRC Press, Boca Raton, USA, 1991), which cannot be performed by sensors of the Clark (L.C. Clark, Electrochemical device for chemical analysis, US Patent 2913386, 1956) type. In comparison to electrochemical sensors fiber optical chemosensors have several advantages, which are illustrated in this paper. Details of the sensor chemistry, instrumental system and the oxygen sensitivity are presented. The application in a precultivation processes, in Ca-alginate beads and layers with immobilized cells in packed-bed-reactors are presented. Diffusion coefficients were calculated from the experiments.

Setup of a fiber optical oxygen multisensor-system and its applications in biotechnology

In bioreactors, metabolically active cells are surrounded by a changing chemical and physical environment. To control such a complex system, the relationship between cells and their environment must be known. For this purpose, information on the regulation process within the cells and between the cells and their environment is important. The cell characteristics, the medium composition, and the physical environment (especially the fluid dynamics of the multiphase system) have to be monitored continuously. Unfortunately, there is still a lack in instrumentation and sensors for continuous monitoring of cell properties during cultivation. The only continuous monitoring technique providing information on the metabolic state of the cells is in situ fluorometry with a miniaturized fluorometer probe. In 1957, Duysens and Amesz first reported on the measurement of one of the key cell metabolites-the nucleotide NAD(P)H-in living microorganisms.’ For this purpose, cell cultures can be irradiated with UV-light, and the NAD(P)H fluorescence light can be detected. In 1970, a special fluorometer was used by Harrison and Chance for the first time to monitor biotechnological processes.’ During the following years, only a few authors reported about applications of this measuring technique in biotechn~logy.~’ In 198 1, Beyeler developed a miniaturized fluorosensor,8 and during the next few years, the interest in this technique increased more and The on-line measurement of NAD(P)H-dependent culture fluorescence gives a direct insight into the microorganisms during cultivation. By adding special tracers to the cultivation medium, fluorometry can also be used for studying the mixing-time behavior in bioreactors. In order to study the culture fluorescence during mixing-time experiments, a miniaturized fluorometer with a special fiber optics for the simultaneous detection of two different wavelengths was constructed and applied to process monitoring and mixing-time experiments. In addition, a commercially available microfluorometer was studied for the measurements of culture fluorescence (Zymomonas mobilis) .

On-line measurement of culture fluorescence for process monitoring and control of biotechnological processes.

Flow cytometric studies during culture of Saccharomyces cerevisiae

Measurement of biological parameters during fermentation processes

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