Many imaging technologies, such as two photon microscopy (TPM), second generation microscopy (SHG) and photoacoustic microscopy (PAM), have successfully demonstrated the ability to extract anatomical, functional and molecular information of biological samples. However, they always fail to obtain comprehensive information from biological tissue due to single imaging modality. To address this limitation, we developed a multimodal microscopy with fully integrated PAM, TPM and SHG. The home-built multimodal microscopy system enable label-free imaging of biomedical tissue with sub-micron resolution in vivo and in vitro. The results may offer a new tool to provide the capable of fast comprehensive information capturing for biological studies.
The kidneys are essential regulatory organs whose main function is to regulate the balance of electrolytes in the blood, along with maintaining pH homeostasis. The study of the microscopic structure of the kidney will help identify kidney diseases associated with specific renal histology change. Spectrally encoded microscopy (SEM) is a new reflectance microscopic imaging technique in which a grating is used to illuminate different positions along a line on the sample with different wavelengths, reducing the size of system and imaging time. In this paper, a SEM device is described which is based on a super luminescent diode source and a home-built spectrometer. The lateral resolution was measured by imaging the USAF resolution target. The axial response curve was obtained as a reflect mirror was scanned through the focal plane axially. In order to test the feasibility of using SEM for depth-section imaging of an excised swine kidney tissue, the images of the samples were acquired by scanning the sample at 10 μm per step along the depth direction. Architectural features of the kidney tissue could be clearly visualized in the SEM images, including glomeruli and blood vessels. Results from this study suggest that SEM may be useful for locating regions with probabilities of kidney disease or cancer.
Spectrally encoded microscopy (SEM) is a new microscopic imaging technique in which a grating is used to illuminate different positions along a line on the sample with different wavelengths, reducing the size of system and imaging time. In this paper, a SEM device is described which is based on a swept source and a balanced detection. A fixed gain balanced detector (BD) was employed in the system for detecting the low sample light without amplifier. Compared to conventional SEM detection method, our BD-SEM device has two significant advantages, one is its capability of suppressing common-mode noise and thermal noise, resulting in the lateral resolution better than direct detection, the other is that it can amplify the signal intensity which is particularly helpful for tissue reflectance imaging. The lateral resolution was measured by imaging a USAF resolution target. The images of onion cells were obtained. The data showed that both the lateral resolution and signal noise ratio are better than non-BD method. The method presented in this work is helpful for developing miniature endoscopic probe for in vivo tissue visualization with high acquisition speed and high imaging quality.
We present a cube data correlation-based correlation mapping optical coherence tomography (cube-cmOCT) method to reconstruct small blood vessel maps. In the cube-cmOCT method, the two adjacent cube data are correlated to extract blood flow information. Both phantom experiments and in vivo experiments were performed to demonstrate the advantage of the proposed method in improving the SNR of blood vessel maps without increasing the window size in the xz plane and offering a clear image of the small blood vessels almost missed by the conventional cmOCT method.
A different real-time self-wavelength calibration method for spectral domain optical coherence tomography is presented in which interference spectra measured from two arbitrary points on the tissue surface are used for calibration. The method takes advantages of two favorable conditions of optical coherence tomography (OCT) signal. First, the signal back-scattered from the tissue surface is generally much stronger than that from positions in the tissue interior, so the spectral component of the surface interference could be extracted from the measured spectrum. Second, the tissue surface is not a plane and a phase difference exists between the light reflected from two different points on the surface. Compared with the zero-crossing automatic method, the introduced method has the advantage of removing the error due to dispersion mismatch or the common phase error. The method is tested experimentally to demonstrate the improved signal-to-noise ratio, higher axial resolution, and slower sensitivity degradation with depth when compared to the use of the zero-crossing method and applied to two-dimensional cross-sectional images of human finger skin.