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Updated: Jul 5, 2025
Single Molecule Fluorescence Microscopy on Planar Supported Bilayers
Published on: October 31, 2015
Motion-resistant three-wavelength spatial frequency domain imaging system with ambient light suppression using an
Yu Feng1, Chen Cao2, Yuto Shimada1
1Shizuoka University, Graduate School of Integrated Science and Technology, Hamamatsu, Japan.
Significance:
We present a motion-resistant three-wavelength spatial frequency domain imaging (SFDI) system with ambient light suppression using an 8-tap complementary metal-oxide semiconductor (CMOS) image sensor (CIS) developed at Shizuoka University. The system addresses limitations in conventional SFDI systems, enabling reliable measurements in challenging imaging scenarios that are closer to real-world conditions.
Aim:
Our study demonstrates a three-wavelength SFDI system based on an 8-tap CIS. We demonstrate and evaluate the system's capability of mitigating motion artifacts and ambient light bias through tissue phantom reflectance experiments and in vivo volar forearm experiments.
Approach:
We incorporated the Hilbert transform to reduce the required number of projected patterns per wavelength from three to two per spatial frequency. The 8-tap image sensor has eight charge storage diodes per pixel; therefore, simultaneous image acquisition of eight images based on multi-exposure is possible. Taking advantage of this feature, the sensor simultaneously acquires images for planar illumination, sinusoidal pattern projection at three wavelengths, and ambient light. The ambient light bias is eliminated by subtracting the ambient light image from the others. Motion artifacts are suppressed by reducing the exposure and projection time for each pattern while maintaining sufficient signal levels by repeating the exposure. The system is compared to a conventional SFDI system in tissue phantom experiments and then in vivo measurements of human volar forearms.
Results:
The 8-tap image sensor-based SFDI system achieved an acquisition rate of 9.4 frame sets per second, with three repeated exposures during each accumulation period. The diffuse reflectance maps of three different tissue phantoms using the conventional SFDI system and the 8-tap image sensor-based SFDI system showed good agreement except for high scattering phantoms. For the in vivo volar forearm measurements, our system successfully measured total hemoglobin concentration, tissue oxygen saturation, and reduced scattering coefficient maps of the subject during motion (16.5 cm/s) and under ambient light (28.9 lx), exhibiting fewer motion artifacts compared with the conventional SFDI.
Conclusions:
We demonstrated the potential for motion-resistant three-wavelength SFDI system with ambient light suppression using an 8-tap CIS.
