Multi-layer holographic reconstruction and focusing strategy-based construction method of spatial distribution of cell group

Abstract

The invention relates to a multi-layer holographic reconstruction and focusing strategy-based construction method of spatial field distribution situations of a cell group, and belongs to the field ofdigital holographic imaging application. The method comprises: setting a starting distance and a termination distance of the reconstruction and step length of each layer, carrying out layer-by-layer reconstruction, and recording a reconstruction situation of each layer; carrying out grayscale counting for a reconstruction image of a certain layer, using lognormal distribution to fit curvature extremums of a curve, setting grayscale thresholds, and combining a morphological processing means to extract a target object region; establishing a cell focusing judgment function, and adding informationof depth, at which focusing is best, after tracking and matching of the same cell imaging centroid between adjacent layers to form spatial positions where cells are located; and respectively calculating focusing depth of each cell in a hologram and horizontal-plane centroid projection in a reconstruction image of a corresponding layer to construct spatial distribution of the cell group. The method lays an important foundation for tracking multi-cell movement trajectories by multi-image holographic reconstruction, and is generally suitable for observation and measurement of microscopic non-cell objects.

Description

technical field

[0001] The invention belongs to the application field of digital holographic imaging, and relates to a cell space field distribution construction method based on multi-layer holographic reconstruction and object focusing strategies. Background technique

[0002] In the actual optical microscopic observation process, since the sample often has a certain thickness, the distribution of cells not only has two-dimensional position information of plane projection, but also has longitudinal depth position information. The strength of cell activity, whether it is apoptotic, or whether it adheres to the wall will cause differences in its spatial distribution. In addition, the focal planes of the optical microscope are distributed in a very narrow range, and it is difficult to ensure that the observed cells in the field of view are well focused at the same time. Even after repeated focusing operations, it is often difficult for observers to gain an intuitive understan...

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