Cell migration is a key feature for living organisms. Image analysis tools are useful in studying cell migration in three-dimensional (3-D) in vitro environments. We consider angiogenic vessels formed in 3-D microfluidic devices (MFDs) and develop an image analysis system to extract cell behaviors from experimental phase-contrast microscopy image sequences. The proposed system initializes tracks with the end-point confocal nuclei coordinates. We apply convolutional neural networks to detect cell candidates and combine backward Kalman filtering with multiple hypothesis tracking to link the cell candidates at each time step. These hypotheses incorporate prior knowledge on vessel formation and cell proliferation rates. The association accuracy reaches 86.4% for the proposed algorithm, indicating that the proposed system is able to associate cells more accurately than existing approaches. Cell culture experiments in 3-D MFDs have shown considerable promise for improving biology research. The proposed system is expected to be a useful quantitative tool for potential microscopy problems of MFDs.
Cell migration studies in 3D environments become more popular, as cell behaviors in 3D are more similar to the behaviors of cells in a living organism (in vivo). We focus on the 3D angiogenic sprouting in microfluidic devices, where Endothelial Cells (ECs) burrow into the gel matrix and form solid lumen vessels. Phase contrast microscopy is used for long-term observation of the unlabeled ECs in the 3D microfluidic devices. Two template matching based approaches are proposed to automatically detect the unlabeled ECs in the angiogenic sprouts from the acquired experimental phase contrast images. Cell and non-cell templates are obtained from these phase contrast images as the training data. The first approach applies Partial Least Square Regression (PLSR) to find the discriminative features and their corresponding weight to distinguish cells and non-cells, whereas the second approach relies on Principal Component Analysis (PCA) to reduce the template feature dimension and Support Vector Machine (SVM) to find their corresponding weight. Through a sliding window manner, the cells in the test images are detected. We then validate the detection accuracy by comparing the results with the same images acquired with a confocal microscope after cells are fixed and their nuclei are stained. More accurate numerical results are obtained for approach I (PLSR) compared to approach II (PCA & SVM) for cell detection. Automatic cell detection will aid in the understanding of cell migration in 3D environment and in turn result in a better understanding of angiogenesis.
Microcirculation lesion is a common symptom of chronic liver diseases in the form of vasculature deformation and circulation alteration. In acute to chronic liver diseases such as biliary atresia, microcirculation lesion can have an early onset. Detection of microcirculation lesion is meaningful for studying the progression of liver disease. We have combined wide-field fluorescence microscopy and a laser speckle contrast technique to characterize hepatic microcirculation in vivo without labeling in a bile-duct ligation rat fibrosis model of biliary atresia. Through quantitative image analysis of four microcirculation parameters, we observed significant microcirculation lesion in the early to middle stages of fibrosis. This bimodal imaging method is useful to assess hepatic microcirculation lesion for the study of liver diseases.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.