Abstract
In this study we present for the first time the use of confocal microscopy and laser scanning brightfield microscopy (LSBF) for real time imaging of femtosecond laser nanosurgery and its dynamics in C. elegans. A single multimodal optical workstation that provides the ability to perform femtosecond laser nanosurgery and simultaneous confocal and LSBF imaging was used for the purpose. With this tool several dynamic phenomena concomitant with laser nanosurgery in C. elegans were observed and imaged. Some of these dynamic phenomena, like muscular contraction and single muscle cell stimulation, have been imaged for the first time during nano-neurosurgery of C. elegans.
Highlights
Light and lasers are increasingly being used as manipulation tools for biological samples and there are several applications that can range from organelle nanosurgery [1], to cell growth stimulation [2]
In this study we present, for the first time to our knowledge, the possibility to simultaneously image highly dynamic phenomena occurring during the femtosecond laser nanosurgery of neuronal axons in C. elegans with a high axial resolution technique such as with confocal microscopy
Laser axotomy was preformed in D-type axonal neurons of C. elegans using MHz femtosecond laser
Summary
Light and lasers are increasingly being used as manipulation tools for biological samples and there are several applications that can range from organelle nanosurgery [1], to cell growth stimulation [2]. There have been significant developments in the technique and recently neuronal processes of few hundred nanometer thickness have been precisely incised without any apparent collateral damage [4]. The most significant being the confinement of induced effects primarily to the focal volume (
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