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: 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: The present invention leverages the techniques for expansion microscopy (ExM) to provide improved high-throughput super-resolution whole-organ imaging methodology to image protein architectures over whole organs with nanoscale resolution by using high-throughput microscopes in combination with samples that have been iteratively expanded more than once, in a method referred to herein as “iterative expansion microscopy” (iExM). In the ExM method, biological samples of interest are permeated with a swellable material that results in the sample becoming embedded in the swellable material, and then the sample can be expanded isotropically in three dimensions The process of iteratively expanding the samples can be applied to samples that have been already expanded using ExM techniques one or more additional times to iteratively expand them such that, for example, a 5-fold expanded specimen can be expanded again 3- to 4-fold, resulting in as much as a 17- to 19-fold or more linear expansion.

Abstract: Various embodiments provide a method of conversion and synthesis of materials via biostructures labeled with an antibody which an inorganic particle has been conjugated. According to various embodiments, a structure corresponding to a biostructure is generated by labeling an antibody which an inorganic particle has been conjugated in a biostructure and growing the inorganic particle with respect to the biostructure.

Abstract: Provided are the nucleic acid-mediated pattern replication and a method of manufacturing a 2-D material using the same. A method of manufacturing a 2-D material according to an embodiment may include preparing a first material having a first nucleic acid patterned on a surface thereof, bonding a linker-nucleic acid to the first nucleic acid, bonding the first nucleic acid and a second nucleic acid attached to a surface of a second material through the linker-nucleic acid and replicating a pattern of the first material to the surface of the second material, separating the first material, and applying a third material on a pattern replicated to the surface of the second material.

Abstract: Various embodiments can provide a method of amplifying a fluorescence signal through the cyclic staining of complementary antibodies conjugated with fluorophores. According to various embodiments, the method of amplifying a fluorescence signal may be configured to prepare a primary antibody and antibody pairs with respect to each target protein, bind the primary antibody to the target protein, and bind the antibody pairs to the primary antibody. The multicolor fluorescence signals may be amplified and fabricated by forming fluorescence signals in a way to prepare different antibodies for which cross reactions with different types of target proteins, respectively, have been removed using an agarose gel and to bind different antibody pairs according to combinations of different antibodies to different primary antibodies bound to target proteins, respectively.

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: The present invention leverages the techniques for expansion microscopy (ExM) to provide improved high-throughput super-resolution whole-organ imaging methodology to image protein architectures over whole organs with nanoscale resolution by using high-throughput microscopes in combination with samples that have been iteratively expanded more than once, in a method referred to herein as “iterative expansion microscopy” (iExM). In the ExM method, biological samples of interest are permeated with a swellable material that results in the sample becoming embedded in the swellable material, and then the sample can be expanded isotropically in three dimensions The process of iteratively expanding the samples can be applied to samples that have been already expanded using ExM techniques one or more additional times to iteratively expand them such that, for example, a 5-fold expanded specimen can be expanded again 3- to 4-fold, resulting in as much as a 17- to 19-fold or more linear expansion.

Abstract: This disclosure relates generally to systems and methods of providing standardized topographical configurations for template regions. In one embodiment, a set of array arrangements is selected. Arrays of template structures are then formed on at least one substrate. Each of the arrays is arranged in accordance with an array arrangement in the set of array arrangements such that the arrays correspond surjectively onto the set of array arrangements. After the arrays are formed, a self-assembly material is provided on the arrays. Self-assembly patterns formed by self-assembling material as a result of the arrays may be empirically observed and used to map a set of self-assembly pattern arrangements surjectively onto the set of array arrangements. Using this mapping, a combination of the self-assembly pattern arrangements that match a target pattern arrangement can be used to select a combination of array arrangements from the set of array arrangements for a template region.

Abstract: This disclosure relates generally to systems and methods of providing standardized topographical configurations for template regions. In one embodiment, a set of array arrangements is selected. Arrays of template structures are then formed on at least one substrate. Each of the arrays is arranged in accordance with an array arrangement in the set of array arrangements such that the arrays correspond surjectively onto the set of array arrangements. After the arrays are formed, a self-assembly material is provided on the arrays. Self-assembly patterns formed by self-assembling material as a result of the arrays may be empirically observed and used to map a set of self-assembly pattern arrangements surjectively onto the set of array arrangements. Using this mapping, a combination of the self-assembly pattern arrangements that match a target pattern arrangement can be used to select a combination of array arrangements from the set of array arrangements for a template region.