In vivo nanosecond laser axotomy: cavitation dynamics and vesicle transport.
Nanosecond laser pulses (lambda = 355 nm) were used to cut mechanosensory neurons in Caenorhabditis elegans and motorneurons in Drosophila melanogaster larvae. A pulse energy range of 0.8-1.2 microJ and < 20 pulses in single shot mode were sufficient to generate axonal cuts. Viability post-surgery was >95% for C. elegans and 60% for Drosophila. Cavitation bubble dynamics generated due to laser-induced plasma formation were observed in vivo by time-resolved imaging in both organisms. Bubble oscillations were severely damped in vivo and cavitation dynamics were complete within 100 ns in C. elegans and 800 ns in Drosophila. We report the use of this system to study axonal transport for the first time and discuss advantages of nanosecond lasers compared to femtosecond sources for such procedures.
National Centre for Biological Sciences-TIFR, Bangalore 560065, India.
G Nageswara
GN
Rao
Sucheta S
SS
Kulkarni
Sandhya P
SP
Koushika
Kaustubh R
KR
Rau
D019771:Axotomy / Q000295:instrumentation*; D001693:Biological Transport, Active / Q000502:physiology; D004867:Equipment Design; D019544:Equipment Failure Analysis; D053685:Laser Therapy / Q000295:instrumentation*; D008866:Microsurgery / Q000295:instrumentation*; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods; D013572:Synaptic Vesicles / Q000502:physiology* / Q000648:ultrastructure*