Micropipette Resonator Enabling Targeted Aspiration and Mass Measurement of Single Particles and Cells.

This paper reports micropipette resonators, mechanical resonator-integrated micropipettes, which enable selective aspiration and mass measurement of particles or cells suspended in liquids with two orthogonal vibration modes. A custom pipette pulling system is built to provide power-modulated linear heating on a rotating glass capillary to make an asymmetric cross section with extended uniformity.A glass capillary is stretched with the custom puller, cut within the pulled region, polished, mounted on a machined metallic jig, and then coated with a metal. As a result, a doubly clamped tube resonator-integrated micropipette is made. For simultaneous frequency readouts of two orthogonal modes, an optical pickup, originally developed for optical data storage, is configured closely above and properly aligned to the micropipette resonator and two digital phase-locked loops are employed. For mass responsivity calibration, frequency shifts of the micropipette resonator are measured with various liquids and glass microparticles. Buoyant masses of unicellular organisms, Paramecium aurelia, freely swimming in a culture dish are successfully measured with two orthogonal modes.

Correlation between thermotolerance and membrane properties in Paramecium aurelia.

The relationship between thermotolerance and membrane properties was studied by using a ciliated protozoan, Paramecium aurelia. P. aurelia is a complex of sibling species termed ;syngens' whose cell morphology appear similar on microscopic examination. From the comparison of tolerance to increasing temperature among 14 syngens of P. aurelia, we selected syngens 2 and 3 as low thermotolerant examples, and syngens 8 and 10 as high thermotolerant examples. The membrane resistance of high thermotolerant syngens measured by injection of a constant inward current was greater than that of low thermotolerant syngens. Membrane fluidity measurements of living cells using the fluorescent dye, 6-lauroyl-2-dimethylaminonaphtalene (laurdan) showed that the fluidity at the cultured temperature was decreased in high thermotolerant syngens compared to that of low thermotolerant syngens. However, when the temperature was increased to the killing temperature of each syngens, the fluidity was increased to almost the same level irrespective of syngen. Furthermore, analysis of fatty acids extracted from whole cells showed that the ratios of unsaturated to saturated fatty acids was smaller in high thermotolerant syngens than in low thermotolerant syngens. These results suggest that the thermotolerance of P. aurelia syngens is determined by the membrane fluidity which is related to the fatty acids composition.