Project 3 (Fraunhofer)
Project 3: Sensitive chip-based assays to investigate physical, chemical and biological factors regarding their influence on the cell biological effects of nanoparticles
Fellow: Abdul Shah (ESR3), 36 months
Tutor: Dr. Yvonne Kohl, Dr. Thorsten Knoll, Dr. Markus Pesch (Fraunhofer)
Within EU research projects we have developed cell based chip technologies for the non invasive characterisation of cells in vitro. Therefore microfluidics for defined medium supply and application of test substances were combined with optical and electrochemical cell characterisation techniques on microchips. Within FP6 DIPNA we have used chip-based technologies and a reporter cell line from partner 1 to analyse sensitively immunomodulatory effects of NP on the single cell level based on the following concept: Cells of a reporter cell line are cultured in micro chambers of a chip. The bottom material of the cell culture chamber is suitable for cell culture, for electron microscopy and for light microscopy. After the application of a defined concentration of NP to the cultured cells, the cell response is determined on the single cell level by quantification of fluorescent protein expression. Afterwards the cells are fixated and the NP are localized by electron microscopy for correlation with the cell response obtained before.
Based on the existing knowledge an ESR in biomedical engineering and a hosted ESR in biology will co-operate in use of sensitive chip-based assays to investigate factors (physical, chemical, biological) regarding their influence on the cell-biological effects of nanoparticles. The ESR focused on bio-engineering will optimize the chip design and experimental setup with respect to the defined application of NP and a more sensitive readout for quantifying expression of fluorescent proteins on the single cell level. For ensuring an application of NP with a known concentration and size distribution the microfluidic of the biochip is combined with a module for light scattering analysis of dispersed NP. The ESR in engineering will be seconded to partner 1 to learn the implementation and interpretation of reporter cell-based immunological assays. Hosted visiting ESRs from partners 1 and 2 will evaluate the behaviour of cells on chip and the results of chip based assays by comparison with standard molecular biological methods like PCR and Western blot. Further for selected nanomaterials the results of chip based assays will be compared to the result obtained by the assays of partner 1. Main skills acquired are engineering of cell-based biochips and application of cell-based biochip technologies for testing nanomaterials. Further the ESR in bio-engineering will acquire basic knowledge in cell and molecular biology. Secondments to partner 1 (reporter gene techniques), 8 (single cell approach for neurons), 12 (characterization of NP parameters under relevant conditions).
In WP1 (tasks 1.1 and 1.3) and WP4 (tasks 4.1 and 4.5).