Abstract

CELLutions Summit Abstract

Use of a Microfluidic Cell Sorting Device for Live-Cell Isolation from a Small Population of Mouse Hypothalamic Neurons

Kurt A. Krummel, Kacee Jones, Haichuan Zhang, Andy Katz, Satchidananda Panda, E. Robert Wassman

To better understand the regulation of circadian oscillator networks, we isolated neurons from a transgenic mouse model that express GFP in neurons of the hypothalamic suprachiasmatic nucleus (SCN). Cells were isolated from experimental and control animals utilizing a microfluidic cell sorter developed by Celula, Inc. Samples were loaded into a disposable, aseptic cartridge that enables sorting with efficient recovery and minimal dilution. We isolated a target population of the brightest 3% of GFP positive neurons from a single animal along with the GFP-dim hypothalamic neurons from the same sample. The resulting viable sorted cells were analyzed by PCR for gene expression analysis of the target neurons. These data will allow the investigation of the properties of these fragile, rare cells with purity not previously attainable. There are two main reasons why our technology enables these experiments for the first time. Each animal has a target population of only a few thousand neurons of interest in the SCN, and high velocity flow rates of traditional cell sorters create shear forces that impact sensitive cells, altering their relevant gene expression. The use of our technology allows previously impossible or impractical isolation of rare, viable, and biologically relevant cells from a variety of sources with high purity and recovery for further characterization including gene expression, or live cell analysis.

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