Seasonal as well as climate changes have immense effect on bud burst, leaf color and leaf abscission. Autumn phenology of leaves is clearly distinguishable in deciduous plant leaves where the leaf color changes from green to red (leaf senescence). In this work, two-photon fluorescence lifetime imaging microscopy (2P-FLIM) and full-field optical coherence microscopy (FF-OCM) were applied to study mitochondrial activity and microstructural changes, respectively, in the senescence of Acer saccharum (Sugar maple) leaves. Fluorescence lifetime of reduced nicotinamide adenine dinucleotide phosphate [NAD(P)H] was recorded using 2P-FLIM to quantify the cellular metabolic changes. Compared to the green leaves, the red leaves showed a 19% increase (P < 0.05) in the average fluorescence lifetime of NAD(P)H, and a 52% decrease (p < 0.005) in the free to protein-bound NAD(P)H ratio. This infers a significant change in mitochondrial metabolic regulation in red leaves in contrast to green leaves. Additionally, en-face sectional images at 0.8 μm axial resolutions of the green and the red color Acer saccharum leaves via FF-OCM using white light emitting diode (WLED) showed a well-defined microstructure of epicuticular waxy layer in green leaves as compared to red leaves where disintegrated microstructure was observed. Our approach can potentially be used to correlate mitochondrial activity with epicuticular microstructural changes in senescing leaves and other biological tissues.
This work reports a Linnik type single shot full-field optical coherence tomography (SS-FF-OCT), which uses a fast generalized analytic signal based complex Riesz transform scheme to reconstruct wide area en-face OCT images. The OCT interferometer is illuminated using a single broad band white light emitting diode (WLED) (wavelength range 470- 850nm, central wavelength 650nm) and detection unit is a two-dimensional (2D) charge complementary metal oxide semiconductor (CMOS) camera. The measured axial and lateral resolution (using 4x, NA=0.1 microscope objective) of the present system is 0.9μm and 3.9μm, very close to theoretical values. The measured imaging area using CMOS camera is 2150 x 2150μm2. The feasibility of the system is demonstrated by imaging scattering samples such as onion bulk and plant leaf. The present SS-FF-OCT is compact, fast (Riesz transform based scheme), stable, cost-effective, and provides comparable axial resolution.
A swept source system was realized in the wavelength range of 810-875 nm with the combination of a broad-band superluminescent diode (SLD) and an acousto-optic tunable filter (AOTF) as a frequency-tuning device. SLD has two spectral centers at 820 nm and 845 nm with spectral bandwidth (FWHM) of around 40 nm. Gaussian spectral shaping
was performed onto the original SLD spectrum while reconstructing OCT images for various test samples such as onion
slice and fingerprint impression taken on a glass substrate. As a pulse can be considered a Gaussian distribution of
frequencies, spectral shaping yields sharper Fourier peaks. Application of Gaussian spectrum facilitates in precisely
locating the reflective boundaries within the sample that results in improved OCT images.
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