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and simultaneous imaging of their activity
(see Fig. 2). The setup uses Thorlabs’ rotat-
able Bergamo multiphoton microscope for
two-photon imaging of brain tissue at a la-
ser wavelength suited to the green fluores-
cent calcium sensors. It can record >1000
neurons in one field of view. For photo-
stimulation of the light-sensitive ion chan-
nel at a different wavelength (1040 nm),
The pattern is achieved by repeatedly
applying a spiral motion of the stimulat-
ing laser focus over the area of interest.
Within this pattern, the field illuminat-
ed by the stimulating laser includes only
neurons that have been genetically modi-
fied and identified (through observation of
their behavioral correlates—for instance,
firing in response to a specific sensory
stimulus) as potentially guiding behavior.
In this way, the neural computation can
be causally probed within a brain region,
and specific activity patterns of certain
neurons can be linked to the behavior-
al output of the animal. The stimulating
laser power, duration, and temporal pat-
tern can be set to drive very well-defined
activity patterns in the targeted neurons.
In vivo photon triggering
of action potentials
With the novel photostimulation and imag-
ing setup, the UCL researchers are pioneer-
ing in the field of targeted optogenetics.
FIGURE 2. Experimental setup with the
Thorlabs Bergamo multiphoton fluorescence
microscope in rotated geometry. In the
background is the BlueCut laser from Menlo
Systems microjoule femtosecond fiber laser
used for photostimulation.
FIGURE 3. The light path of the stimulation
laser, with spatial modulator that shapes the
beam into a pattern that effectively activates
the target population of neurons, indicated by
yellow circles shown in the figure on page 39.
FIGURE 1. The all-optical setup highlights
the interplay between behavioral experiment,
imaging of activity patterns in the brain, and
the manipulation of specific functionally