We demonstrate a method that reduces the noise caused by multi-scattering (MS) photons in an \invivo optical coherence tomography image. This method combines a specially designed image acquisition (i.e., optical coherence tomography scan) scheme and subsequent complex signal processing. For the acquisition, multiple cross-sectional images (frames) are sequentially acquired while the depth position of the focus is altered for each frame by an electrically tunable lens. In the signal processing, the frames are numerically defocus-corrected, and complex averaged. Because of the inconsistency in the MS-photon trajectories among the different electrically tunable lens-induced defocus, this averaging reduces the MS signal. This method was validated using a scattering phantom and in vivo unanesthetized small fish samples, and was found to reduce MS noise even for unanesthetized in vivo measurement.

pdf/multiple-scattering-suppression-for-in-vivo-optical-2kj2oug9wb.pdf

Multiple scattering suppression for in vivo optical coherence tomography measurement using B-scan-wise multi-focus averaging method

Small teleosts have recently been established as models of human diseases. However, measuring heart rate by electrocardiography is highly invasive for small fish. The physiological nature and function of vertebrate autonomic nervous system (ANS) modulation of the heart has traditionally been investigated in larvae with an incompletely developed ANS or in anesthetized adults, whose ANS activity may possibly be disturbed under anesthesia. Here, we defined the frequency characteristics of heart rate variability (HRV) modulated by the ANS from observations of heart movement in high-speed movie images and changes in ANS regulation under environmental stimulation in unanesthetized adult medaka (Oryzias latipes), a small teleost. The HRV was significantly reduced by atropine (1 mM) in the 0.25 – 0.65 Hz and by propranolol (100 μM) at 0.65–1.25 Hz range, suggesting that HRV in adult medaka is modulated by both the parasympathetic and sympathetic nervous systems within these frequency ranges. Such modulations of HRV by the ANS were remarkably suppressed in anesthetized adult medaka. Continuous exposure to light suppressed HRV only in the 0.25 – 0.65 Hz range, indicating parasympathetic withdrawal. The power of HRV increased along developmental processes. These results suggest that ANS modulation of the heart in adult medaka is frequency-dependent phenomenon, and that the impact of long-term environmental stimuli on ANS activities can be precisely evaluated in unanesthetized adult fish using this method.

Acquirement of the autonomic nervous system modulation evaluated by heart rate variability in medaka (Oryzias latipes)

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Acquirement of the autonomic nervous system modulation evaluated by heart rate variability in medaka (Oryzias latipes)