Automated Phototransfection

Filed in by on September 11, 2014 • views: 321
Automated Phototransfection

This project is on automating the phototransfection process on neuron and fibroblast cells. Phototransfection is presently done manually in a very tedious manner: First, the cell of interest needs to be identified in the microscope FOV and location recorded. Then an appropriate location of where to administer a laser beam on the cell body needs to be determined. Applying the laser to the wrong location can damage the cell. The laser beam creates a hole in the cell membrane where donor mRNA can be inserted to the cell from a pipette, completing the process. A framework for fully automating this procedure has been designed and proof-of-concept implementation achieved. Computer vision techniques are used to identify the cell of interest in the FOV and determine target locations for the laser beam. A control program takes this information and coordinates movements of the computer controlled XY stage, translating the coordinates of the laser target location to a predefined, fixed, laser firing location. A 20X improvement was shown to be possible with this implementation with room for improvement to greater than 80X.

Selected Publications:
  1. D. Cappelleri, A. Halasz, J-Y. Sul, T. Kim, J. Eberwine, and V. Kumar, “Towards Fully Automated Phototransfection”, Proceedings of the IEEE Conference on Robotics, Automation, and Science (CASE), Bangalore, India, August 22-25, 2009.*
  2. D. Cappelleri, A. Halasz, J-Y. Sul, T. Kim, J. Eberwine, and V. Kumar, “Towards A Fully Automated High-Throughput Phototransfection System”, Journal of the Association for Laboratory Automation (JALA), vol. 15, pp. 329-341, August 2010, DOI information: 10.1016/j.jala.2010.03.003.
  3. J-Y Sul, C. K. Wu, F. Zeng, J. Jochems, M. T. Lee, T. K. Kim, T. Peritz, P. Buckley, D. Cappelleri, M. Maronski, M. Kim, V. Kumar, D. Meaney, J. Kim and J. Eberwine, “Transcriptome Transfer Produces a Predictable Cellular Phenotype”, Proceedings of the National Academy of Science, 2009: May 5;106(18):7624-9. Epub 2009 Apr 20. PMCID: PMC2670883.