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Glutaraldehyde enhanced dielectrophoretic yeast cell separation
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FIG. 1.

Dual-shell oblate spheroid model for a yeast cell, where and represent the spheroid primary and secondary axes, respectively.

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FIG. 2.

calculated from the dual-shell oblate spheroid model plotted against frequency for viable and nonviable yeast cells.

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FIG. 3.

The fabricated quadrupole electrode array when activated can produce (a) pDEP or (b) nDEP.

Image of FIG. 4.

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FIG. 4.

Effects of cross-linking on the high and low cofs on viable yeast cells as a function of media conductivity for viable yeast cells. (a) Low frequency cof. (b) High frequency cof. GLT has a minimal effect on viable yeast cell electrical properties. (c) Low frequency cof. (d) High frequency cof. Unlike viable cells, GLT has a large effect on nonviable yeast cell electrical properties.

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FIG. 5.

[(a) and (b)] Calculated effects of GLT on viable yeast cells. (a) Calculated cell wall conductivity. The observed peak relates to optimal buffer conditions where the cell wall ion channels are most open. (b) Cytoplasm conductivity. The slight difference over the active ion channel range is most likely due to cytoplasm leakage into the media. [(c) and (d)] Calculated effects of cross-linking on nonviable yeast cells. (c) Cell wall conductivity. (d) Cytoplasm conductivity.

Image of FIG. 6.

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FIG. 6.

Comparison of for [(a) and (b)] uncross-linked and [(c) and (d)] cross-linked cells. (a) Low frequency crossover over media conductivity range for viable and nonviable cells. A small gap between measured crossover (possible separation) exists between 150 and . (b) Differences in cof observed for the high frequency crossover. Separation is possible over larger conductivity range . (c) The observed low frequency crossover difference is much larger when the cells are cross-linked. (d) High frequency crossover data: cells have a large range of media conductivity where cell separation is possible.

Image of FIG. 7.

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FIG. 7.

Effects of GLT on viable (transparent) and nonviable (blue) DEP separation in cells are suspended in media conductivity of with a field frequency of 2.5 MHz. (a) Untreated 1:1 cell mixture indicates poor separation. (b) GLT treated cell mixture has improved DEP separation over the untreated sample.


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We introduce a method for improved dielectrophoretic (DEP) discrimination and separation of viable and nonviable yeast cells. Due to the higher cell wall permeability of nonviable yeast cells compared with their viable counterpart, the cross-linking agent glutaraldehyde (GLT) is shown to selectively cross-link nonviable cells to a much greater extent than viable yeast. The DEP crossover frequency (cof) of both viable and nonviable yeast cells was measured over a large range of buffer conductivities in order to study this effect. The results indicate that due to selective nonviable cell cross-linking, GLT modifies the DEP cof of nonviable cells, while viable cell cof remains relatively unaffected. To investigate this in more detail, a dual-shelled oblate spheroid model was evoked and fitted to the cof data to study cell electrical properties. GLT treatment is shown to minimize ion leakage out of the nonviable yeast cells by minimizing changes in cytoplasm conductivity over a large range of ionic concentrations. This effect is only observable in nonviable cells where GLT treatment serves to stabilize the cell cytoplasm conductivity over a large range of buffer conductivity and allow for much greater differences between viable and nonviable cell cofs. As such, by taking advantage of differences in cell wall permeability GLT magnifies the effect DEP has on the field induced separation of viable and nonviable yeasts.


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752b84549af89a08dbdd7fdb8b9568b5 journal.articlezxybnytfddd
Scitation: Glutaraldehyde enhanced dielectrophoretic yeast cell separation