Application of an advanced maximum likelihood estimation restoration method for enhanced-resolution and contrast in second-harmonic generation microscopy

What is it about?

Second-harmonic generation imaging is an emerging microscopic tool to understand molecular organization of unlabeled biological structures such as collagen. Conventionally, visualizing them using a two photon microscopy is a gold standard in understanding their dynamics at a resolution theoretically limited by the diffraction of light. However, due to scattering, absorption and changes in refractive indices it is hard to reach that theoretical resolution limit. Here we show a custom-built iterative image restoration algorithm titled advanced maximum likelihood algorithm (AdvMLE), that uses deconvolution to restore blurred photons to their original state. From the restored images we recovered high spatial frequencies using Fourier analysis. The AdvMLE processed images of chicken tendon collagen and mouse heart sarcomeres shows substantial improvement in both lateral and axial resolution, which helps in visualizing and analyzing the structures with better precision.

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Why is it important?

We believe that this algorithm could be modified in future to restore images from other optical modalities where a conventional resolution improvement is not possible such as magnetic resonance imaging (MRI) or computer assisted tomography (CATScan), and obtain better precision in diagnosis and treatment.

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