GitHub

Evaluation of the robustness and accuracy of PCA-based algorithms for in-line Digital Holographic Microscopy

In this undergraduate research project, we aim to investigate the performance of the PCA-based method for phase reconstruction in an in-line DHM system. Specifically, we will evaluate the accuracy and robustness of PCA-based phase reconstruction algorithms based on the number of phase-shifted images and the value of the phase step. The PCA algorithm will be tested under both noiseless and noisy conditions to assess its reliability in practical imaging scenarios. This research project seeks to contribute to the understanding of the potential of PCA as a viable method for phase reconstruction in DHM and its applicability in real-world imaging applications.

Classification of olive oil blends by image processing RGB images

This MATLAB code enables the indirect measurement of the chlorophyll molecules present when a green laser diode illuminates the oil sample. Oil blends can be classified into five classes (no olive oil, light olive oil, medium olive oil, olive oil, and extra virgin olive oil) by quantifying the ratio of the red channel versus the green channel along the laser illumination path from a color image.

In-focus quantitative phase imaging from defocused off-axis holograms: synergistic reconstruction framework

Digital Holographic Microscopy (DHM) enables the three-dimensional (3D) reconstruction of quantitative phase distributions from a defocused hologram. We have developed a synergistic computational framework to simultaneously compensate for the linear tilt introduced in off-axis DHM systems and focus the defocused holograms by minimizing a cost function, providing in-focus reconstructed phase images without phase distortions. 

More information and Github repository at: https://github.com/OIRL/3D-QPI-DHM/ 

Documentation: https://sites.google.com/view/3dqpidhm/home 

Semi-heuristic phase compensation algorithm

The proposed Semi-heuristic phase compensation (SHPC) method is an alternative approach for reconstructing phase maps with reduced processing time compared with the brute-force algorithms while avoiding the local-minimum problem of the heuristic proposals. This algorithm is based on nested searches in which the grid size in every iteration is systematically reduced to optimize the compensation time. This algorithm also have a dynamic version called D-SHPC for compensating holographic videos of dynamic samples. Both algorithms are accurate in phase reconstructions and fast enough to compensate full FOV (1280x960 pixels) holograms at rates of 5 FPS.

Global Stress Detection Framework

 We have proposed a global stress detection framework combining eight HRV features and an RF model with a classifying power of 99% or higher during individual testing using two benchmark datasets (i.e., WESAD and SWELL datasets). The proposed global stress detection framework is available on GitHub. We used Google Collab with Python 3 to investigate the proposed global stress framework. The code was implemented using Python libraries such as TensorFlow, Matplotlib, NumPy, Pandas, and Karas. The computational specs for the algorithm are 12 GB RAM using Google Collaboratory.

Open-access tool for Phase Compensation Reconstruction Method in Digital Holographic Microscopy Operating in Non-Telecentric Regime

Among the optical configuration of the imaging system in DHM, imaging systems operating in a non-telecentric regime are the most common ones. The spherical wavefront introduced by the non-telecentric DHM system must be compensated to provide undistorted phase measurements. Here, we present a clear and simple procedure for a total compensation algorithm for non-telecentric DHM holograms independent of the sample's size (i.e., no requiring sample-free field of view within the hologram). The proposed method has been implemented in MATLAB 2021a and Python 3.7.1. Users of the proposed method should have installed the Optimization and Global Optimization MATLAB toolboxes and the scipy library from Python.

More information and Github repository at: https://github.com/OIRL/noteleDHM-Tool 

Documentation: https://sites.google.com/view/noteledhmtool/home?authuser=1

YouTuve video: www.youtube.com/watchv=Hyi5vwgYX7chtps://youtu.be/Hyi5vwgYX7

Design and evaluation of a confocal scanning microscope using off-of-shelf optical components by means of Zemax OpticStudio optical design software

Confocal microscopes are known for optically removing the out-of-focus information (i.e., blur) in each transverse section of the sample’s volume, providing a more accurate three-dimensional image of thick microscopic samples than widefield microscopes. In this work, we have designed and evaluated a confocal microscope using off-of-shelf optical components from Thorlabs’ catalog, one of the major optical manufacturers. The design and evaluation have been implemented using Zemax OpticStudio, the standard optical system design software. Here, we have also reported a practical protocol for building a confocal microscope using sequential mode.

More information and Github repository at: https://oirl.github.io/Design-Confocal.github.io/

Design and evaluation of a spectrometer using off-of-shelf optical components by means of Zemax OpticStudio optical design software

Spectrometers are optical tools to measure the spectral composition of a beam of light. Key features of an optimal spectrometer are: cost, variation of the spot size along the spectral range, spot size compared to the diffraction limit, and minimum presence of aberrations. Here, we present different optical designs to focus the light emerging from a diffraction grating.

More information and Github repository at: https://oirl.github.io/Design-espectometer.github.io/

Mueller Matrix from intensity-based images recorded by a bright-field microscope

The hallmark of polarization-sensitive microscopy is the estimation of polarimetric information of samples, making them a suitable tool for detecting and diagnosis different type of cells and tissues in life sciences. In this work, we describe a practical procedure for measuring the Mueller matrix of samples using a bright-field microscope using 36 intensity-based images. The hallmark of the proposed method is the retrieval of the Mueller matrix across the transverse section of a sample, being suitable for analyzing biological samples. The method has been implemented in Python and MATLAB.

More information and Github repository at: https://oirl.github.io/Muller-Matrix-Microscopy/

pyDHM Library

pyDHM is an open-source Python library aimed at Digital Holographic Microscopy (DHM) applications. The pyDHM is a user-friendly library written in the robust programming language of Python that provides a set of numerical processing algorithms for reconstructing amplitude and phase images for a broad range of optical DHM configurations. The pyDHM implements phase-shifting approaches for in-line and slightly off-axis systems and enables phase compensation for telecentric and non-telecentric systems. In addition, pyDHM includes three propagation algorithms for numerical focusing complex amplitude distributions in DHM and digital holography (DH) setups. We have validated the library using numerical and experimental holograms.

More information and Github repository at: https://catrujilla.github.io/pyDHM/


Examples of how to install and use it are found in our YouTube channel.

pyDHM library structure. The library is composed of utility (1), phase-shifting (2), fully-compensated phase reconstruction (3), and numerical propagation (4) packages.

Blind-PS-DHM-methods

We present two blind-iterative phase-shifting algorithms in which there is no need for prior knowledge of the phase shifts between the raw holograms. These two approaches provide accurate quantitative phase images in Phase-Shifting Digital Holographic Microscopy (PS-DHM) using three or two raw holograms. The proposed methods are based on the demodulation of the spectral components of the recorded holograms. The hallmarks of our methods are blindness (no prior knowledge of any phase shift), accuracy, and reduced acquisition and processing times leading to a PS-DHM system more suitable for dynamic imaging, as is the case of live cell imaging and colloidal systems.  

More information and Github repository at: https://oirl.github.io/Blind-PS-DHM-methods/ 

Exaple of the phase reconstructions with the two algorithms.

Speckle-Hybrid-median-mean

We present a single-shot computational method based on the use of a hybrid median-mean approach for reducing the speckle noise. The proposed method can be applied to both amplitude and phase reconstructed images. This method is based on the combination of multiple median-filtered images with different kernel sizes. Because, for each median-filtered image the speckle position changes, the average of these median-filtered images results in a final image with low speckle contrast and no resolution decrease (e.g., no blurring effect introduced by the median filter). The proposed method has been evaluated experimentally in digital holography and digital holographic microscopy.

More information and Github repository at: https://oirl.github.io/Speckle-Hybrid-median-mean/ 

Example of the result of denoising process with the algorithm.

Learning-based cGAN method for QPI for off-axis DHM systems

The conventional quantitative phase reconstruction in off-axis Digital Holographic Microscopy (DHM) relies on computational processing. Regardless the implementation, any DHM computational processing leads to large processing times, hampering the use of DHM to video-rate renderings of dynamic biological processes. In this work, a conditional generative adversarial network (cGAN) for robust and fast quantitative phase imaging in DHM is reported. The reconstructed phase images provided by the GAN model present stable background levels, enhancing the visualization of the specimens for different experimental conditions. The proposed learning-based method has been trained and validated using human red blood cells. After proper training, the proposed GAN yields to a computationally efficient method, reconstructing DHM images 7X faster than conventional computational approaches.

More information and Github repository at: https://oirl.github.io/cGAN-Digital-Holographic-microscopy/

Example of the result of the CGAN model

tuDHM

tuDHM is an algorithm to recover the complex object information for a single-shot digital holographic microscope (DHM) operating in the telecentric regimen. The algorithm is based on the minimization of a cost function that finds the best numerical conjugated reference beam to compensate the filtered object information, eliminating any undesired phase perturbation due to the tilt between the reference and object waves

More information and Github repository at: https://oirl.github.io/tuDHM/ 

Graphical flowchart of the algorithm