Laser Capture Microdissection of Drosophila Peripheral Neurons

Laser capture MICRODISSECTION or LCM has emerged as an extremely powerful tool that can be used to isolate specific cell types from tissue sections with a high degree of spatial resolution and accuracy. Here LCM is used to dissect peripheral neurons from third instar oph larvae. Third instar larvae are embedded in a cryo mold and frozen then a cryo status used to cut serial tissue sections, which are placed on glass slides thawed and fixed in 70%ethanol.

The slides are then incubated with tripsin to remove residual larval cuticles and subjected to dehydration via ethanol gradient and xylene clearing. Next an LCM cap is placed on the tissue section and selective fluorescent neuron cells are captured using a pulsing laser. Finally, RNA is isolated from the captured cells for downstream applications such as quantitative R-T-P-C-R or microarray analyses.