The Andor ALC-UVP-350i is a computer controlled UV pulsed laser source and optical configuration for use in micro-ablation of sub-cellular organelles and molecular uncaging. The instrument is part of an Andor Revolution laser spinning disk confocal system installed at 'The London Centre for Nanotechnology'.
Dichroic UV objective adapter allows the UV pulses from the fiber (left) to be focused into the specimen. The adapter, from Meopta, has a long pass nature and an edge at 405 nm.
The UVP-350i uses a Spectra Physics 350 nm, 60 μJ solid state device and provides computer control of pulse energy, repetition rate (1 Hz to 5kHz) and number of pulses delivered. Inside the unit, a beam attenuator and a series of ND optical filters control pulse energies in the range ~ 60 nJ to 6 pJ – covering the ranges used for ablation and uncaging. UV pulses are delivered to the microscope via the custom dichroic adapter shown on the right. This adapter is manufactured by Meopta and specified by Andor for UV operation. UV pulses are transmitted to the microscope via a multi-mode UV optical fiber. The fiber core diameter is 25 μm and can be focused into the specimen via a specialized dichroic adapter and microscope objective, as shown on the right. The UV beam diameter in the specimen plane is ~3μm, providing precise targeting.
NB - Preliminary results below, show the effective use of the instrument for the target application.
Cultured neurons in phase contrast – shows before and after ablation of a neurite within the red circle. Circle diameter approximately 10 μm. Pulse energy level was set to 10 nJ and a single pulses was required to ablate the process shown.
Mouse B16 Melanoma cells in phase contrast – shows before and after ablation within the red circle. Circle diameter approximately 10 μm. Pulse energy level was set to 10 nJ and 5 pulses @ at a rate 5kHz were required to ablate the process shown.
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