MICROFLUIDIC MAGNETIC CELL SORTINGSYSTEM FOR CANCER DIAGNOSIS :MICROFLUIDIC MAGNETIC CELL SORTINGSYSTEM FOR CANCER DIAGNOSIS Biochip Technology Seminar Session Murugan Veerapandian 2008-40152 03/Mar/2009


OUTLINE :OUTLINE Introduction to magnetic microfluidic system for cell sorting and analysis Theory What authors did? Results and discussion Advantages My view on this microfluidic diagnostic strategy


INTRODUCTION TO MAGNETIC MICROFLUIDIC SYSTEM FOR CELL SORTING AND ANALYSIS :INTRODUCTION TO MAGNETIC MICROFLUIDIC SYSTEM FOR CELL SORTING AND ANALYSIS Smart system for the diagnosis of rare malignant cells like circulating tumor cells (CTC) Array of magnetic columns coated with antibodies, which has membrane specific protein for target cell capture were taken place inside the chip After trapping in the magnetic columns thier morphology and expression were analysed from microscopy


THEORY :THEORY Immunoaffinity separation matrix made by magnetic beads and its self assembly in the column (Magnetic bead flow) Key ref:biomedcentral.inist.fr/images/1477-044X-1-2-1.jpg


WHAT AUTHORS DID? :WHAT AUTHORS DID? Detection of B-lymphocytes expressing human protein CD19 membrane protein (CD19+) mixed with T- lymphocytes (CD19-) Cell tracker CMFDA dye The capture of epithelial cells (MCF7 cell line) spiked in a mixture of lymphocytes The beads are grafted either with anti-CD19 (antibody) or anti-EpCAM antibody, depending on the targeted population of cells Cells are observed in bright field and in fluorescent light at high magnification Key ref: NATURE. Vol.450, 20/27, 2007


SCHEMATIC REPRESENTATION OF CELL SEPARATION METHOD AND MICROFLUIDIC CHIP :SCHEMATIC REPRESENTATION OF CELL SEPARATION METHOD AND MICROFLUIDIC CHIP (a) Photography of the microfluidic chip. Magnetic beads are injected in channel 2, the mixture of cells in channel 1 and rinsing buffer in channel 3. The channel 4 is the output of the system. (b) The large section of the separation chamber (50µm in height and 1cm in width) allows a significant throughput. The input channel is divided in a “tree structure” that provide a homogeneous flowdistribution in the separation chamber. (c) Hexagonal arrays of 10 µm diameter ferrofluid spots are formed by microcontact printing.


RESULTS AND DISCUSSION :RESULTS AND DISCUSSION In chip immunostaining and May-Grunwald Giemsa coloration of lymphocytes B cells (Raji). (a) Bright field image. (b) Staining with anti-CD19-FITC antibody. (c) Hoechst staining. (d) Merge image of (b) and (c). (e) May-Grünwald-Giemsa staining Cell mixture with 1 positive cell per 1000 negative were studied Small sample size of 0.5 mL/h Magnetic array is directly stamped on a coverslip, which is compatible with modern spectroscopy for high throughput cell sorting and analysis


ADVANTAGES :ADVANTAGES Cheaper in fabrication Disposable cell sorting device Requires smaller sample sizes Useful in immunology and cancer research


MY VIEW :MY VIEW Evaluation of new class DNA interactive molecule for drug screening application In vitro device for targeted drug deilvery binding study Early diagnosis of autoimmune and neurodegenearaive disorders (A- ß amyloid protofibrils in Alzheimer’s disease, mutant a-synuclein linked early onset of Parkinson’s disease, etc.)


REFERENCES :REFERENCES Saias et.al., 552-554, (µTAS2008) Nagrath et al., Nature, 450, 1235 (2007) Doyle et al., Science, 295, 2237 (2002) Fütterer et al., Lab Chip, 4, 351-356 (2004) and www.fluigent.com


Slide 11:Thank you for your kind attention Got question?