Isolation of Neurons from Caenorhabditis elegans via Fluorescence-Activated Cell Sorting
Transcrição
Take a suspension of cells obtained from transgenic Caenorhabditis elegans worms in a culture medium.
The suspension contains cells from various tissues, including neurons from the nervous system.
The transgenic worms were engineered to express a fluorescent protein conjugated to a vesicular transmembrane transporter. This transporter, present on synaptic vesicles carrying the neurotransmitter acetylcholine, is found in cholinergic neurons throughout the body.
Centrifuge the cell suspension and discard the supernatant. Resuspend the cells in a fresh culture medium for fluorescence-activated cell sorting, or FACS.
Analyze the cell suspension using FACS. The fluorescently labeled cholinergic neurons emit fluorescence upon exposure to light of an appropriate wavelength, separating them from other unlabeled cells.
Transfer the isolated neurons into a multi-well plate. Incubate the cells in a humidified chamber to maintain the moisture level.
