Lab-on-a Chip Systems. with Optical Detection of Toxicological Effects. in Medicine

Transcription

1 Lab-on-a Chip Systems with Optical Detection of Toxicological Effects in Medicine Karl-Heinz Feller Ernst-Abbe-Fachhochschule Jena Jencolor Innovation Forum Jena

2 RESEARCH GROUP INSTRUMENTAL ANALYTICS supervisor: Prof. Dr. Karl-Heinz Feller 5 PhD students, 3 Post-Doc, 3 scientific co-workers, student research assistants

3 Detection of toxic/irritating effects Development of a cell-based sensorchip for the complex description of the physiological property of a substance towards keratinocytes (HaCaT cell line) in a real time and sensitive manner to be applied within the cosmetic and pharmaceutical industry. Determination of toxic effects: - strong reaction (apoptosis) - weak reaction (cellular stress, irritation) - no reaction Development of a fast method based on a lab-on-achip by application of early biomarker 8. Jahrestagung AK BioMST

4 Detection of toxic/irritating effects test substance microscopy cell density cell adhesion supervision fluorescence cellular stress level promoter reporter gene GFP detection Possibility to differentiate between competing, stress- or growth-related effects cell model electrochemistry cell morphology cell damage impedance spectroscopy detection

5 Detection of toxic/irritating effects test substance fluorescence cellular stress level promoter reporter gene GFP detection Possibility to differentiate between competing, stress- or growth-related effects cell model electrochemistry cell morphology cell damage impedance spectroscopy detection

6 Establishment of the fluorescence-based assay Measurement of fluorescence intensity in dependance on the cellular stress level Identification of a powerful biomarker for detection of cellular stress Cloning Stable Transfection Real-Time quantitative PCR Genomic DNA Establishment of a reporter assay PCR Selection by G418 Implementation of the fluorescence- based assay on the lab-on-a-chip

7 Setup of the Lab-on-a-Chip micro mixer test substance cell culture media reference substance keratinocyte Lab-on-a-Chip

8 Setup of the Lab-on-a-Chip micro mixer test substance cell culture media reference substance stress keratinocyte promoter reporter gene green fluorescent protein Lab-on-a-Chip

9 Setup of the Lab-on-a-Chip micro mixer test substance cell culture media APD detection reference substance stress keratinocyte temperature control electrochemical readout promoter reporter gene green fluorescent protein Lab-on-a-Chip control and data analysis

10 Incubator-independent cultivation Conventional cell culture practice work flow microscope limitation CO 2 - incubator No online-monitoring CO 2 -incubator for optimal culture conditions condition temperature 37 C constant ph HCO H 3 O + <-> CO 2 + 2H 2 O high humidity solution transparent heater CO 2 -independent medium sterile cultivation under flow conditions (flow rate < 10µl/min) 8. Jahrestagung AK BioMST

11 MICROFLUIDICS & MICROSENSORS Systematic built-up of a measuring system with LEDs and multi-color sensors

12 FLUID DYNAMICS SIMULATION WITH ANSYS CFX CFD enables an optimization before the manufacturing process Micromixer improve the homogenity, selectivity and cost efficiency investigation and combination of different micromixer modules

13 NUMERICAL SIMULATIONS fluid flow simulation of mixing and multi- phase- systems fluid- structure interaction simulation of e.g. valve s, pumps etc.

14 OPTICAL SIMULATION FRED is used for numerical simulation of optical processes to optimize the opt. Yield as well as to minimize optical losses

15 DETECTORS true-colour/multicolor Sensor (MAZeT): color regions per sensor (Δλ~50 nm) Mini-Spectrometer (Ocean Optics): - small, high spectral resolution (Δλ=0,5 nm) CCD Camera: - high sensitivity, special resolution Avalanche photodiode: - highest sensitivity, small

16 SENSOR PRINCIPLE with MIPs Glycoprotein Multicolour Sensor MIP Optical Fiber Light source CCD-Camera Sample Micromixer Chip-cuvette with spotted MIP s Spectrometer Solvent

17 NANOTOXISCREEN Lab-on-a-Chip-based detection of the cell damaging potential of nano particels on human cells

18 Conclusions and outlook - Development of a lab-on-a-chip system for the complex analysis of cytotoxic effects of chemical substances - no incubator necessary (flow-through cultivation of keratinocytes) - CFD-optimization of the mixing process and the shear stress - Control of the growing cell culture - Combination of optical and electrochemical detection unit - Automatically monitoring of the cell behavior - Integration of the complex analysis in a three-channel-chip - low-cost option for the optical detection by means of avalanche photodiodes 18

19 Thank you very much for your attention!!! Funding: BMWi (ZIM-Programm), EU (EFRE, ITN), BMBF, u. a.