KEYWORDS: Collagen, Second-harmonic generation, Interference (communication), Calibration, Skin, In vivo imaging, Statistical analysis, Signal to noise ratio, Optical fibers, Biological research
It is commonly believed that intrinsic skin aging is
associated with the change of the collagen structures.
The influence of the diabetes on the skin collagen is
also considered to be similar to aging. Moreover,
second-harmonic-generation (SHG) in collagen fibers is
known to reflect the detailed collagen structures. It is
thus highly valuable to adopt the SHG intensity as a
collagen structure indicator. With the help of SHG,
recently one can achieve in vivo imaging which
provides the information of what really happens
beneath the human skin. However, when analyzing the
images, the SHG brightness of each pixel highly
depends on the illumination condition, the depth of the
SHG source, and the voltage of PMT. Therefore, it is
important to calibrate these factors before statistical
analysis. In this paper, we present our recent
development that calibrates the in vivo SHG images by
using noises. We first determine the regions of signals
and noises by setting a threshold relating to the standard
deviation of the image. By using the assumption that
the noise was amplified by PMT with an amplification
ratio the same as the SHG signal, we can define the
brightness of the noise region as a parameter
representing the voltage of PMT, and use this parameter
to calibrate all SHG images. After calibrating, we can
then compare different images from volunteers and
analyze the influence of aging and diabetes on the SHG
intensity from collagen fibers, even if the voltage of
PMT was not fixed.
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