On-chip non-invasive and label-free cell discrimination by impedance spectroscopy.

OBJECTIVES: Many flow-cytometric cell characterization methods require costly markers and colour reagents. We present here a novel device for cell discrimination based on impedance measurement of electrical cell properties in a microfluidic chip, without the need of extensive sample preparation steps and the requirement of labelling dyes. MATERIALS AND METHODS, RESULTS: We demonstrate that in-flow single cell measurements in our microchip allow for discrimination of various cell line types, such as undifferentiated mouse fibroblasts 3T3-L1 and adipocytes on the one hand, or human monocytes and in vitro differentiated dendritic cells and macrophages on the other hand. In addition, viability and apoptosis analyses were carried out successfully for Jurkat cell models. Studies on several species, including bacteria or fungi, demonstrate not only the capability to enumerate these cells, but also show that even other microbiological life cycle phases can be visualized. CONCLUSIONS: These results underline the potential of impedance spectroscopy flow cytometry as a valuable complement to other known cytometers and cell detection systems.

Stray-light effects of diffractive beam-shaping elements in optical microsystems.

The use of diffractive beam-shaping elements in hybrid or monolithic microsystems is investigated. Compact optical systems require diffractive structures with small grating periods for creating large deflection angles. Such elements are difficult to fabricate while a low stray-light level is maintained. In addition, because of the small geometrical dimensions and the short propagation lengths in an optomechanical microsystem, any stray light generated by the diffractive structure critically affects the overall optical performance. A model for the estimation of the interference effects between the designed and the unwanted diffraction orders is developed and applied to an example of a collimating diffractive optical element. On the basis of theoretical and experimental results, design rules for the application of diffractive beam-shaping elements in microsystems are derived.

Analysis and optimization of fabrication of continuous-relief diffractive optical elements.

The fabrication of continuous-relief diffractive optical elements by direct laser beam writing in photoresist is analyzed. The main limitation and tolerances are identified, and their influence on optical performance is quantified. Fabricated structures show rounded profile steps resulting from the convolution of the desired profile with the writing beam. This leads to a reduction in diffraction efficiency. Optimization techniques are presented to minimize this effect. Scaling the profile depth by a factor of mu > 1 increases the first-order diffraction efficiency for blazed elements. This method is also applied to suppress the zeroth diffraction order in computer-generated holograms. A nonlinear compensation of the exposure data for the Gaussian beam convolution results in an 18% increase of the diffraction efficiency for a blazed grating with a 10-mum period to a value of 79%.