Abstract: Multiplexed fluorescent imaging which is essential for finding out how various biomolecules are spatially distributed in cells or tissues is disclosed. The present disclosure may obtain 10 or more different biomolecule images with one labeling and imaging by newly designing selection of fluorophores, detection spectral ranges, and signal unmixing algorithm. The present disclosure is a blind unmixing technology for unmixing an image without an emission spectrum of fluorophore, and in this technology, 4 pairs of fluorophores are used, and each pair consists of two fluorophores in which emission spectra are overlapped. Each pair of fluorophores is strongly excited by only one excitation laser. Two images with different detection spectral ranges are obtained for each pair, and two images are unmixed via mutual information minimization without fluorophore emission spectrum information. Two images also may be unmixed via Gram-Schmidt orthogonalization and fluorescence measurement based unmixing.

Abstract: The present disclosure relates to a method and device for unmixing fluorescent signals based on a joint histogram. The method and device may be configured to obtain a plurality of images of a plurality of fluorescent materials marked in different biomolecules and to unmix the signals of the fluorescent materials based on at least one joint histogram index defined between the images of each of pairs each including two or more of the obtained images. According to the present disclosure, the joint histogram index may include at least one of Kullback-Leibler divergence, cross-entropy, or Rand-index.

Abstract: Disclosed are three-dimensional image registration via dense staining and a super-resolution multiplexed imaging method using the same. The three-dimensional image registration includes: placing at least one fiducial marker in an internal structure of a living tissue via dense staining; obtaining a plurality of images by repeatedly imaging the living tissue while replacing a fluorescent label attached to at least one target molecule within the living tissue where the fiducial marker is placed; and registering the images using the fiducial marker as a reference to obtain a final image of the target molecule.

Abstract: This disclosure provides a method and apparatus for multicolor unmixing through iterative mutual information minimization in a simultaneous channel update structure. In this disclosure, a plurality of images may be obtained with respect to a plurality of fluorophores labeled on different biomolecules, respectively. Images of the fluorophores may be unmixed from the obtained images, respectively, while simultaneously minimizing mutual information shared between the obtained images based on all permutation combinations of the obtained images. According to the present disclosure, signals having three or more fluorophores whose emission spectra are overlapped in one spectral range can be more accurately unmixed.

Abstract: The present disclosure provides a method and apparatus for biomolecular multiplexed imaging through the iterative unmixing of fluorophores. According to the first embodiments of the present disclosure, although signals of two fluorophores are detected in the first fluorescent detection spectral range, the signals of the two images obtained from two detection spectral ranges can be unmixed through the iterative minimization of mutual information. Furthermore, the present disclosure provides a multi-color unmixing method and apparatus through the iterative minimization of mutual information. In the second embodiments of the present disclosure, a plurality of images of a plurality of fluorophores marking different biomolecules, respectively, for example, N fluorophores are obtained. Images, each containing the signals of single fluorophore can be obatined from the obtained images while minimizing the mutual information shared by images of each of pairs each consisting of two of the obtained images.

Abstract: Multiplexed fluorescent imaging which is essential for finding out how various biomolecules are spatially distributed in cells or tissues is disclosed. The present disclosure may obtain 10 or more different biomolecule images with one labeling and imaging by newly designing selection of fluorophores, detection spectral ranges, and signal unmixing algorithm. The present disclosure is a blind unmixing technology for unmixing an image without an emission spectrum of fluorophore, and in this technology, 4 pairs of fluorophores are used, and each pair consists of two fluorophores in which emission spectra are overlapped. Each pair of fluorophores is strongly excited by only one excitation laser. Two images with different detection spectral ranges are obtained for each pair, and two images are unmixed via mutual information minimization without fluorophore emission spectrum information. Two images also may be unmixed via Gram-Schmidt orthogonalization and fluorescence measurement based unmixing.

Abstract: A light field microscope may record a raw light field video of a sample. The raw video recording may be decomposed into a non-negative low-rank component and a non-negative sparse component. The low-rank component may correspond to a static portion of the sample, and the sparse component may correspond to a dynamically changing portion of the sample. Volume reconstruction may be performed on the sparse component to generate a three-dimensional video of the sample, with improved spatial resolution. In some cases, the decomposition is calculated by an alternating direction method of multipliers algorithm, with the non-negativity of the sparse component and low-rank component enforced after each iteration. In some cases, the volume reconstruction is calculated by Richardson-Lucy iteration with regularization. The sample may be fluorescent. The fluorescence may be indicative of neural activity in the sample.

Abstract: The information budget of a light field microscope is increased by increasing the field of view and image circle diameter of the microscope, while keeping the ratio of overall magnification of the microscope to the numerical aperture of the microscope unchanged. Alternatively, the information budget is increased by increasing the field of view and image circle diameter of the microscope by a first factor, while increasing the ratio of overall magnification of the microscope to the numerical aperture of the microscope by a smaller, second factor. In some cases, an infinity-corrected light field microscope has an overall magnification that is greater than the nominal magnification of the objective lens.

Abstract: The information budget of a light field microscope is increased by increasing the field of view and image circle diameter of the microscope, while keeping the ratio of overall magnification of the microscope to the numerical aperture of the microscope unchanged. Alternatively, the information budget is increased by increasing the field of view and image circle diameter of the microscope by a first factor, while increasing the ratio of overall magnification of the microscope to the numerical aperture of the microscope by a smaller, second factor. In some cases, an infinity-corrected light field microscope has an overall magnification that is greater than the nominal magnification of the objective lens.

Abstract: The information budget of a light field microscope is increased by increasing the field of view and image circle diameter of the microscope, while keeping the ratio of overall magnification of the microscope to the numerical aperture of the microscope unchanged. Alternatively, the information budget is increased by increasing the field of view and image circle diameter of the microscope by a first factor, while increasing the ratio of overall magnification of the microscope to the numerical aperture of the microscope by a smaller, second factor. In some cases, an infinity-corrected light field microscope has an overall magnification that is greater than the nominal magnification of the objective lens.

Abstract: The information budget of a light field microscope is increased by increasing the field of view and image circle diameter of the microscope, while keeping the ratio of overall magnification of the microscope to the numerical aperture of the microscope unchanged. Alternatively, the information budget is increased by increasing the field of view and image circle diameter of the microscope by a first factor, while increasing the ratio of overall magnification of the microscope to the numerical aperture of the microscope by a smaller, second factor. In some cases, an infinity-corrected light field microscope has an overall magnification that is greater than the nominal magnification of the objective lens.

Abstract: The information budget of a light field microscope is increased by increasing the field of view and image circle diameter of the microscope, while keeping the ratio of overall magnification of the microscope to the numerical aperture of the microscope unchanged. Alternatively, the information budget is increased by increasing the field of view and image circle diameter of the microscope by a first factor, while increasing the ratio of overall magnification of the microscope to the numerical aperture of the microscope by a smaller, second factor. In some cases, an infinity-corrected light field microscope has an overall magnification that is greater than the nominal magnification of the objective lens.

Abstract: The information budget of a light field microscope is increased by increasing the field of view and image circle diameter of the microscope, while keeping the ratio of overall magnification of the microscope to the numerical aperture of the microscope unchanged. Alternatively, the information budget is increased by increasing the field of view and image circle diameter of the microscope by a first factor, while increasing the ratio of overall magnification of the microscope to the numerical aperture of the microscope by a smaller, second factor. In some cases, an infinity-corrected light field microscope has an overall magnification that is greater than the nominal magnification of the objective lens.