Correlative holographic fluorescence microscopy for single-shot volumetric imaging and applications thereof

A system comprising:

• an optical imaging tool configured to perform fluorescence microscopy and holographic microscopy;
• and
• a processor and a non-transitory computer readable medium operably coupled thereto, the processor operationally coupled and configured to control the optical imaging tool and to acquire data from the optical imaging tool, wherein the non-transitory computer readable medium comprising a plurality of instructions stored in association therewith that are accessible to, and executable by, the processor, to perform one or more operations, which comprise:
• acquiring, via the optical imaging tool, a fluorescence image and a digital hologram of a sample comprising one or more objects;
• generating a two-dimensional depth map of the one or more objects based on the digital hologram;
• correlating the two-dimensional depth map of the one or more objects with the fluorescence image of the sample;
• and
• generating a volumetric image of the one or more objects in a three-dimensional volume.

A method, comprising:

• acquiring a fluorescence image and a digital hologram of a sample comprising one or more objects;
• generating a three-dimensional map of the one or more objects based on the digital hologram;
• generating a two-dimensional depth map from the three-dimensional map;
• generating a binary segmentation map from the fluorescence image for one or more fluorescence colors;
• generating a volumetric image of the one or more objects based on the two-dimensional depth map and the binary segmentation map;
• and
• outputting the volumetric image to a display.