Abstract
Photostimulation has been instrumental in the past two decades for studying the structural synaptic plasticity and functional connectivity of neuronal circuits. With the advent of optogenetic strategies, this approach has been further expanded and used to identify the neuronal substrates of behavior via monitoring and modulating the activity of specific neuronal types in vivo. To date, however, photostimulation has been mainly implemented via full-field illumination and laser scanning protocols, which suffer from limited selectivity and stop short of generating asynchronous and spatially distributed neuronal firing patterns, characteristic for brain activity.
In this chapter, we discuss advances in using novel light patterning techniques which allow shaping illumination to create flexible spatiotemporal photostimulation profiles over large ensembles of neurons, as well as onto subcellular compartments. Specifically, we describe two light patterning strategies implemented through intensity and phase modulation, respectively. We illustrate the underlying physical principles, their applications to date, and the scope and limitations of each method, in an attempt to bridge the gap between the development of optical techniques and their use for neuroscience experiments.
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Acknowledgement
We would like to thank Aurélien Bègue, Adriana ‘Dăbȃcan’, Priyanka Gupta, and Matthew Koh for constructive critique and comments. This work was supported by an EMBO Long-Term Fellowship and a Swartz Foundation Fellowship (FA), and CSHL start-up funds and a Louis and Gertrude Feil Foundation Fellowship (DFA).
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Glossary
Glossary
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1.
4f configuration : an optical configuration involving two lenses of focal length ‘f’. The object is placed at focal length distance from the first lens and the distance between the two lenses is 2f. This configuration ensures that the distance between the object and its image is 4f. Even when two lenses of different focal lengths are used (f1 and f2), the configuration is still commonly referred to as a 4f. system.
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Absorption cross-section: a proportionality constant that determines the probability of a photon being absorbed by an absorber molecule. It has the units of area, which is why it is known as cross-section and can be imagined to be a perfectly absorbing disk of that area.
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Collimated beam: a beam of light with near-zero divergence that propagates through a given medium. Lasers are highly collimated, but light from extended sources like lamps and LEDs is not.
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Comb function: a periodic function composed of individual delta functions repeated at a particular interval. Also known as impulse train or sampling function in engineering. The Fourier transform of the comb function is another comb function with different periodicity.
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Conjugate plane: if all points residing in a given plane P are imaged onto another plane P’ by a lens, then P and P’ are said to be conjugate planes of each other. In an optical system with more than one lens, changes in the amplitude or phase at any given plane propagate to all other conjugated planes.
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Diffraction grating: an optical component with periodic variation of phase across the surface that causes constructive/destructive interference to produce characteristic diffraction patterns.
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Diffraction-limited spot: the theoretical minimum spot size that can be achieved after focusing a coherent light beam using a lens. The size of the spot at the focal plane is proportional to the wavelength of light and inversely proportional to the numerical aperture of the lens. The function that describes the intensity distribution of a diffraction-limited spot in 3D is known as the point-spread function.
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Focal plane: the point where rays parallel to the optical axis converge after passing through a lens is called the focus. The plane perpendicular to the optical axis containing the focus is called the focal plane (technically the front focal plane). Back focal plane refers to the image plane of an object placed at infinity and is located at a focal distance from the center of the lens, but symmetrically opposite from the front focal plane.
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Gaussian spot: a spot generated by a Gaussian beam when focused, whose intensity profile can be fitted with 2D Gaussian function in the lateral plane.
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Ghost-replicas: the higher-order diffraction patterns (low-intensity repeats) of a light mask when using a coherent light source.
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LED (Light-emitting diodes): a small semiconductor device made up of a pn-junction diode that emits photons (whose energy correspond to the band gap) when an electric potential is applied.
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Mean free path: the average distance travelled by light (photons) between successive collisions (scattering) in the propagating medium.
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Rectangular (rect) function: a step function of a particular duration. Its Fourier transform is a sinc function, whose un-normalized form matches the diffracting pattern from a single-slit (rect function) experiment.
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Temporal coherence: two waves are coherent if they have a constant phase difference between them. For light sources, it is a condition where all the individual light emitters (electron transitions in atoms at the source) have a constant phase difference between themselves, as is the case of LASERs.
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Wavefront: the spatial envelope of all the points in a wave that have the same phase.
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Anselmi, F., Banerjee, A., Albeanu, D. (2015). Patterned Photostimulation in the Brain. In: Douglass, A. (eds) New Techniques in Systems Neuroscience. Biological and Medical Physics, Biomedical Engineering. Springer, Cham. https://doi.org/10.1007/978-3-319-12913-6_9
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