What is it about?
We generate an extended focus image of a sample of transparent algae, using a video of holograms. An extended focus image is an image composed of all the objects in the view field in focus, even when in the original image the object is out of focus. In this work, we use a new criterion for the focus plane detection of each transparent object (phase object). In the simulation process, we use a K-means clustering algorithm for reducing the number of positive cell detection. In the experimental process, we noise interferes with the k-means algorithm.
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Why is it important?
The detection of focus plane of transparent objects is hard to accomplish, because of the interference fringes that are produced at the out of focus planes in this case. Using a hologram the complex field of the sample is retrieve and many slices of the sample are calculated. The criterion analyzes in the transversal and longitudinal directions the reconstructed slices. The analysis is done in the amplitude field. And the algorithm is applied autonomously to the video.
Perspectives
The analysis of dynamic cells is a challenging task because most of the cells are transparent, use a colorant could change the cell behavior. Methods as phase contrast microscope have the limitation of a narrow focus depth, that is, only a cell plane is in focus at the time. Holographic microscopy allows to record all the object information and reconstruct the object in different planes even in out of focus planes. We present a method that could help to find the focus plane of all the cell in the view field of the camera using only one hologram. The method could be applied in the study of living cells.
Dr Maria Luisa Cruz Lopez
Universidad Panamericana Campus Guadalajara
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Autonomous generation of extended images of dynamic phase objects in a depth volume sample using a simple focusing criterion and K-means clustering
, June 2017, SPIE,
DOI: 10.1117/12.2270359.
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