Characterizing refractive index and thickness of biological tissues using combined multiphoton microscopy and optical coherence tomography.

We present a noninvasive method for characterizing the refractive index (RI) and thickness distribution in biological tissues using a combined multiphoton microscopy (MPM) and optical coherence tomography (OCT) system. Tissue layers are distinguished by the MPM and OCT images, and the RI and thickness of each layer are determined by analyzing the co-registered MPM and OCT images. The precision of this method is evaluated on four standard samples which are water, air, immersion oil and cover glass. Precision of within ~1% error compared to reference values is obtained. Biological tissue measurement is demonstrated on fish cornea. Three layers are detected, which are identified as the epithelium and stroma I and II of the cornea. The corresponding RI of each layer is measured to be ~1.446-1.448, 1.345-1.372, and 1.392-1.436, respectively. The difference of RI in the three layers correlates with the tissue compositions including cells in epithelium, large collagen fiber bundles in stroma I, and small collagen fibers in stroma II. The combined MPM/OCT technique is shown to be able to distinguish tissue layers through biochemically specific contrasts and measure RI and thickness of tissue layers at different depths.

Multiscale multimodal imaging with multiphoton microscopy and optical coherence tomography.

A multiscale multiphoton microscopy (MPM) and optical coherence tomography (OCT) system has been developed using a sub-10 fs Ti:sapphire laser. The system performs cross-sectional OCT imaging over millimeter field-of-view and en-face high-resolution MPM imaging with submicrometer resolution from the same sample location. With fish cornea, we have demonstrated cross-sectional imaging of cornea tissue layers using OCT, and the zoom-in imaging of cells and collagen fibers in each layer using MPM. The multiscale MPM/OCT system shows the potential of a rapid coarse scan to search for abnormal regions and the subsequent fine zoom-in imaging for diagnosis.

Selection of convolution kernel in non-uniform fast Fourier transform for Fourier domain optical coherence tomography.

Gridding based non-uniform fast Fourier transform (NUFFT) has recently been shown as an efficient method of processing non-linearly sampled data from Fourier-domain optical coherence tomography (FD-OCT). This method requires selecting design parameters, such as kernel function type, oversampling ratio and kernel width, to balance between computational complexity and accuracy. The Kaiser-Bessel (KB) and Gaussian kernels have been used independently on the NUFFT algorithm for FD-OCT. This paper compares the reconstruction error and speed for the optimization of these design parameters and justifies particular kernel choice for FD-OCT applications. It is found that for on-the-fly computation of the kernel function, the simpler Gaussian function offers a better accuracy-speed tradeoff. The KB kernel, however, is a better choice in the pre-computed kernel mode of NUFFT, in which the processing speed is no longer dependent on the kernel function type. Finally, the algorithm is used to reconstruct in-vivo images of a human finger at a camera limited 50k A-line/s.

High-speed spectral domain optical coherence tomography using non-uniform fast Fourier transform.

The useful imaging range in spectral domain optical coherence tomography (SD-OCT) is often limited by the depth dependent sensitivity fall-off. Processing SD-OCT data with the non-uniform fast Fourier transform (NFFT) can improve the sensitivity fall-off at maximum depth by greater than 5dB concurrently with a 30 fold decrease in processing time compared to the fast Fourier transform with cubic spline interpolation method. NFFT can also improve local signal to noise ratio (SNR) and reduce image artifacts introduced in post-processing. Combined with parallel processing, NFFT is shown to have the ability to process up to 90k A-lines per second. High-speed SD-OCT imaging is demonstrated at camera-limited 100 frames per second on an ex-vivo squid eye.