High throughput lens-free three-dimensional tracking of sperm

摘要

A system for three dimensional imaging of motile objects includes an image sensor and a sample holder disposed adjacent to the image sensor. A first illumination source is provided and has a first wavelength and positioned relative to the sample holder at a first location to illuminate the sample. A second illumination source is also provided having a second wavelength, different from the first wavelength, and positioned relative to the sample holder at a second location, different from the first location, to illuminate the sample. The first and second illumination sources are configured to simultaneously, or alternatively, sequentially illuminate the sample contained within the sample holder. Three dimensional positions of the motile objects in each frame are obtained based on digitally reconstructed projection images of the mobile objects obtained from the first and second illumination sources. This positional data is connected for each frame to obtain 3D trajectories of motile objects.

主权项

1. A system for three dimensional imaging of motile objects contained within a sample comprising: an image sensor; a sample holder configured to hold the sample, the sample holder disposed adjacent to the image sensor; a first illumination source having a first wavelength and positioned relative to the sample holder at a first location to illuminate the sample; a second illumination source having a second wavelength, different from the first wavelength, and positioned relative to the sample holder at a second location, different from the first location, to illuminate the sample, and wherein both the first illumination source and the second illumination source illuminate the sample from the same side of the sample holder; at least one digital processor configured to receive a plurality of image frames generated by the image sensor; and wherein the first illumination source and the second illumination source are configured to simultaneously illuminate the sample contained within the sample holder over a period of time and wherein the at least one digital processor receives the plurality of image frames and identifies holographic projections and microscopic images of the motile objects from the first illumination source and the second illumination source and reconstructs a three dimensional trajectory of the motile objects observed over the period of time based on the acquired holographic projections and classifies the three dimensional trajectory of the motile objects into one or more types of trajectories.