Micromanipulator systems, TransferMan 4r and FemtoJet 4i

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Abstract In the parent grant, we plan to learn the state-of-the-art micromanipulation system in Dr. Marko's lab at Northwestern University and then set up our own one at the University of Illinois, Urbana-Champaign. By using the micromanipulation system in Marko's lab, we have obtained many interesting data on meiotic chromosomes and published one paper in Communications Biology. In addition, we are the first group who isolated metaphase I spindle with chromosomes in mouse oocytes. This will provide novel approaches to study chromosome segregation and abnormality. Our findings on inter-chromosomal interaction will also establish a new paradigm on chromosome segregation. My local co-investigator, Dr. Leckband, has a similar micromanipulation system at the University of Illinois, Urbana-Champaign. However, this system lacks automatic micromanipulators. Without the two manipulators, it is impossible to capture the meiotic chromosomes and further measure their biophysical properties. After setting up the system locally, more members from my lab can be trained to use the cutting-edge technology, which is essential for the success of these proposed projects. We recently found that the measurement of meiotic chromosomes from mutant spermatocytes and oocytes could not be conducted in Dr. Marko's lab because fresh mutant testes and ovaries are required. The mutant spermatocytes became unhealthy after the shipment from Urbana to Evanston. However, all the mutant mouse lines are currently maintained at the University of Illinois, Urbana-Champaign. This means we have to use a local micromanipulation system to measure the stiffness of the chromosomes from the mutant spermatocytes and oocytes. The data collected from these mutants is fundamental for the proposed projects. In summary, we are the first group to apply the micromanipulation system onto meiotic chromosomes. Thus, the local system will be vital for completion of the proposed projects, dramatically speed up these projects, and facilitate intriguing discoveries.