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Investigating biomechanical noise in neuroblastoma cells using the quartz crystal microbalance.

ABSTRACT: Quantifying cellular behaviour by motility and morphology changes is increasingly important in formulating an understanding of fundamental physiological phenomena and cellular mechanisms of disease. However, cells are complex biological units, which often respond to external environmental factors by manifesting subtle responses that may be difficult to interpret using conventional biophysical measurements. This paper describes the adaptation of the quartz crystal microbalance (QCM) to monitor neuroblastoma cells undergoing environmental stress wherein the frequency stability of the device can be correlated to changes in cellular state. By employing time domain analysis of the resulting frequency fluctuations, it is possible to study the variations in cellular motility and distinguish between different cell states induced by applied external heat stress. The changes in the frequency fluctuation data are correlated to phenotypical physical response recorded using optical microscopy under identical conditions of environmental stress. This technique, by probing the associated biomechanical noise, paves the way for its use in monitoring cell activity, and intrinsic motility and morphology changes, as well as the modulation resulting from the action of drugs, toxins and environmental stress.

SUBMITTER: Prasad A

PROVIDER: S-EPMC4424671 | biostudies-literature | 2015 May

REPOSITORIES: biostudies-literature

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Project description:High percentages of harmful microbes or their secreting toxins bind to specific carbohydrate sequences on human cells at the recognition and attachment sites. A number of studies also show that lectins react with specific structures of bacteria and fungi. In this report, we take advantage of the fact that a high percentage of microorganisms have both carbohydrate and lectin binding pockets at their surface. We demonstrate here for the first time that a carbohydrate nonlabeled mass sensor in combination with lectin-bacterial O-antigen recognition can be used for detection of high molecular weight bacterial targets with remarkably high sensitivity and enhanced specificity. A functional mannose self-assembled monolayer in combination with lectin concanavalin A (Con A) was used as molecular recognition elements for the detection of Escherichia coli W1485 using a quartz crytsal microbalance (QCM) as a transducer. The multivalent binding of Con A to the E. coli surface O-antigen favors the strong adhesion of E. coli to the mannose-modified QCM surface by forming bridges between these two. As a result, the contact area between cell and QCM surface that increases leads to rigid and strong attachment. Therefore, it enhances the binding between E. coli and the mannose. Our results show a significant improvement of the sensitivity and specificity of the carbohydrate QCM biosensor with a experimental detection limit of a few hundred bacterial cells. The linear range is from 7.5 x 10(2) to 7.5 x 10(7) cells/mL, which is four decades wider than the mannose-alone QCM sensor. The change of damping resistances for E. coli adhesion experiments was no more than 1.4%, suggesting that the bacterial attachment was rigid, rather than a viscoelastic behavior. Little nonspecific binding was observed for Staphylococcus aureus and other proteins (fetal bovine serum, Erythrina cristagalli lectin). Our approach not only overcomes the challenges of applying QCM technology for bacterial detection but also increases the binding of bacteria to their carbohydrate receptor through bacterial surface binding lectins that significantly enhanced specificity and sensitivity of QCM biosensors. Combining carbohydrate and lectin recognition events with an appropriate QCM transducer can yield sensor devices highly suitable for the fast, reversible, and straightforward on-line screening and detection of bacteria in food, water, and clinical and biodefense areas.

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Investigating biomechanical noise in neuroblastoma cells using the quartz crystal microbalance.

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