Abstract
Stimuli to excitable cells and various cellular processes can cause cell surface deformations; for example, when excitable cell membrane potentials are altered during action potentials. However, these cellular changes may be at or below the diffraction limit (in dendrites the structures measured are as small as 1 µm), and imaging by traditional methods is challenging. Using dual lenses incoherent holography lattice light-sheet (IHLLS-2L) detection with holographic phase imaging of selective fluorescent markers, we can extract the full-field cellular morphology or structural changes of the object’s phase in response to external stimulus. This approach will open many new possibilities in imaging neuronal activity and, overall, in light sheet imaging. In this paper, we present IHLLS-2L as a well-suited technique for quantifying cell membrane deformation in neurons without the actuation of a sample stage or detection microscope objective.
Highlights
Excitable cells, whether neurons or muscle cells, rapidly transport ions during excitation [1]
Lattice Light Sheet Microscopy provides a method for the generation of 3D imaging using a self-reinforcing lattice of Bessel beams as an excitation source and imaging the subsequent fluorescence orthogonally
The lattice light sheet is formed by projecting a linearly polarized circular laser beam stretched in the x axis with cylindrical lenses and compressed in the z axis onto a spatial light modulator (SLM)
Summary
Whether neurons or muscle cells, rapidly transport ions during excitation [1]. Depolarization of the plasma membrane is achieved by cation flux, including Na+ and K+ and Ca2+ ions These processes lead to depolarization, and in excitable cells the explosive activation of voltage gated Na+ channels to cause an action potential. At cell and tissue scale, resolutions in biomedicine, digital holography is used to provide experimental access to the quantitative phase of various cell types and tissues [19,20,21,22]; for example, to reconstruct blood cell movements [15,16], and to perform microsurgeries on cells [17] All these techniques use coherent light, such as laser, which introduces extra noise from speckle and spurious interference that limit the imaging capabilities.
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