34 results about "Subcellular organelle" patented technology

The invention relates to the technical field of image processing, and discloses a stem cell microscopic image feature extraction method and system. The method comprises the following steps: carrying out segmentation processing on a stem cell delay 3D confocal microscopic image through a chromatin texture sensitive adaptive threshold segmentation algorithm to obtain a nucleoplasm separation mask image; performing feature extraction processing on the marker fluorescence image according to the mask image to obtain a radial distribution feature vector and a pluripotency maintenance index; performing time sequence quantification processing on the state change sequence based on the index to obtain differentiation track feature data; performing spatial feature extraction processing on the subcellular organelle region through a topological graph construction algorithm to obtain a spatial correlation network graph; and performing feature fusion processing through a multi-layer perceptron fusion algorithm to obtain a stem cell image feature classification result. The accuracy of stem cell state recognition and the reliability of differentiation trajectory prediction are improved.

The invention discloses a Yarrowia lipolytica engineering bacterium for producing astaxanthin in a peroxisome compartment and application of the Yarrowia lipolytica engineering bacterium. A complete astaxanthin synthesis path is expressed in host bacteria by recombinant Yarrowia lipolytica; comprising geranyl geranyl diphosphate synthase CrtE, phytoene synthetase / lycopene cyclase CrtYB, phytoene desaturase CrtI, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR, beta-carotene ketolase CrtW and beta-carotene hydroxylase CrtZ, and the enzymes are relocated into peroxisome, so that the enzyme activity of the peroxisome is improved, and the enzyme activity of the peroxisome is improved. The number of expression hemoglobin genes VHb and peroxisome is increased, the surface area of the hemoglobin genes VHb is enlarged, and one or more of enzymes PEX11, PEX19, DNM1 or PHO85 for prolonging the service life of the hemoglobin genes VHb and peroxisome is / are obtained; the astaxanthin production performance of the recombinant strain is verified based on cytoplasm engineering and subcellular organelle compartment engineering, and the astaxanthin production capacity of the yarrowia lipolytica is further improved.

The present application relates to a kind of electron donor (alpha-trithiophene) modified rhodamine dye, its synthesis method and application.The dye has strong absorption of 500-600 nm, can emit efficiently in 550-700 nm band, suitable for deep tissue imaging.The structure endows mitochondria targeting ability, can be used as biomarker probe to realize subcellular organelle accurate positioning;Meanwhile, under light, it can efficiently produce reactive oxygen species through type I mechanism, suitable for the photodynamic regulation and biosensing for non-disease diagnosis and treatment purposes.In addition, the dye can also be used as organic photocatalyst for environmental or energy-related photocatalytic reaction.The present application provides a new choice for multi-scene optoelectronic functional materials.

The invention relates to the technical field of biological engineering, and discloses a method for enhancing the urea circulation function of a 3D biological printing liver organ, which comprises the following steps: a cell preparation step: through a genetic engineering technology, enabling mitochondria of liver cells to express a light-driven proton pump, and enabling intracellular expression of a fluorescent probe for reporting ammonia concentration; an organ construction step: mixing the cells with hydrogel to prepare bio-ink, and constructing a three-dimensional liver organ by using a 3D biological printing technology; a system integration step: placing the liver organ in an integrated photoelectric regulation and control system capable of monitoring a fluorescence signal in real time and applying illumination; and a closed-loop enhancement step: the system monitors the ammonia concentration in real time, and when the ammonia concentration exceeds a preset threshold value, illumination is automatically applied to activate the proton pump, so that the urea circulation function is enhanced as required. Through closed-loop light control and a subcellular organelle energy channel, on-demand, efficient and high-temporal-spatial-resolution dynamic enhancement of the liver organ urea circulation function is achieved.

The invention relates to the technical field of molecular probes, in particular to a pH response type AIE molecular probe capable of regulating subcellular organelle targeting and a preparation method and application of the pH response type AIE molecular probe, and the structural general formula of the pH response type AIE molecular probe comprises one of a formula (I), a formula (II) and a formula (III). According to the present invention, the molecular probe has the AIE characteristic group, such that the molecular probe can emit strong fluorescence in the aggregation state, and when the low-concentration molecular probe and the HeLa cell are co-incubated, the cell imaging can be only performed within 30 min; meanwhile, the structure of the molecular probe is modified by utilizing A, C, G and U, so that specific targeting mitochondria or lysosome can be regulated and controlled, and the defects of poor biocompatibility and the like are overcome; meanwhile, the molecular probe is good in selectivity and high in biocompatibility, related instruments are convenient to use, and the molecular probe is a high-sensitivity detection means in current chemiluminescence detection methods.

The invention discloses a peroxisome positioning signal peptide and application thereof. The invention relates to an application of an N-terminal II-type peroxisome positioning signal peptide alone or in combination with a C-terminal I-type signal peptide in the biosynthesis of a compartment high-value compound. The invention comprises the construction of an auxiliary carrier of two peroxisome signal peptides, and covers the application of the auxiliary carrier in the biosynthesis of terpene substances such as limonene, linalool and patchouli alcohol, and flavonoid substances such as daidzein, genistein, 3-hydracrylic acid and coenzyme Q10. The peroxisome signal peptide disclosed by the invention can provide a new element for metabolic engineering modification of subcellular organelles of rhodotorula expansa.

The application discloses a kind of nanoclusters with golgi targeting and its preparation method and application, the nanocluster is zinc sulfide nanocluster, the surface of the zinc sulfide nanocluster is coupled with targeting ligand, the targeting ligand can actively target RA synovial activated macrophage, and can target golgi subcellular organelle, the nanocluster can release H2S gas in inflammatory weak acid microenvironment.The application can realize "inflammatory joint enrichment-> macrophage active uptake-> lysosome escape-> golgi deep anchoring" multi-level precise delivery, release H2S gas in inflammatory weak acid microenvironment, fundamentally reshape intracellular Ca 2+ Steady state, relieve endoplasmic reticulum, golgi and mitochondria triple stress, block inflammatory signal pathway, realize the efficient, safe treatment of RA.

The present application relates to a multi-color fluorescence fluctuation imaging system and method based on a slowly varying transmittance dichroic mirror, which comprises: using multiple wavelength quantum dot fluorescence to mark different subcellular organelle structures, and using a single wavelength excitation light to obtain multi-color fluorescence signals; using a slowly varying transmittance dichroic mirror to split the multi-color fluorescence signals, and using a single camera to simultaneously collect multi-color fluorescence signals in the reflection channel and the transmission channel; using a method based on high-order correlation function calculation to separate different wavelength fluorescence signals; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using a multi-wavelength channel signal mutual iteration method to reduce cross-color between different channels; using

The present application relates to the field of image processing technology and discloses a method and system for extracting features from stem cell microscopic images. The method comprises: segmenting a time-lapse 3D confocal microscopic image of a stem cell using a chromatin texture-sensitive adaptive threshold segmentation algorithm to obtain a nuclear-cytoplasmic separation mask image; performing feature extraction processing on a marker fluorescence image based on the mask image to obtain a radial distribution feature vector and a pluripotency maintenance index; performing temporal quantization processing on a state change sequence based on the index to obtain differentiation trajectory feature data; performing spatial feature extraction processing on a subcellular organelle region using a topological map construction algorithm to obtain a spatial correlation network map; and performing feature fusion processing using a multi-layer perceptron fusion algorithm to obtain a stem cell image feature classification result. The present application improves the accuracy of stem cell state recognition and the reliability of differentiation trajectory prediction.

The invention discloses an ultra-stable red fluorescent protein. The fluorescent protein disclosed by the invention is a protein with an amino acid sequence SEQ ID No.4. Experiments prove that the fluorescent protein has excellent electron microscope sample preparation resistance, can retain more fluorescent signals after osmic acid treatment, and can tolerate embedding of Epon resin; meanwhile, the fluorescent protein has excellent thermal stability, and can retain more fluorescent signals at 90 DEG C; in addition, the fluorescent protein has high chemical stability and light stability. Based on the related properties of the fluorescent protein, the fluorescent protein can be used in the fields of protein labeling, various types of fluorescence imaging and super-resolution photoelectric correlated imaging, and is used for tracking the structural and morphological changes of samples such as proteins, subcellular organelles, local cell regions and cells; the kit can be used for packaging related viruses or transgenic animals, marking specific proteins, specific organelles or specific cells, and realizing rapid transparent tissue imaging and expansion super-resolution imaging.

A method for detecting labile zinc (Zn2+) in a biological material is provided herein, the method including: (a) contacting the biological material with a composition including NapBu-BPEA; and (b) imaging the biological material via molecular fluorescence imaging to detect the labile zinc in the biological material. Also provided herein are methods for morphology-correlated detection of labile zinc localization in a subcellular organelle of a living cell and methods for tracking a change in labile zinc localization in a biological material.

The invention provides preparation of an ultra-small nano probe with efficient biomarker performance and super-resolution imaging application of the ultra-small nano probe. The nanodot is of a core-shell structure, and the core-shell structure comprises a fluorescent core, a fluorescent layer and a fluorescent layer, wherein the fluorescent core is composed of a single-chain semiconductor polymer; the shell layer is composed of an amphiphilic compound, a hydrophobic part of the amphiphilic compound is combined with the semiconductor polymer, a hydrophilic group is located on the surface of the shell layer to serve as an active group, the average particle size of the nanodot is smaller than 5 nm, and the nanodot is of a glassy structure. The nanodot can realize high-density specific marking of various subcellular organelles and nano-scale biological fluorescence imaging.

The invention discloses saccharomyces cerevisiae engineering bacteria for efficiently synthesizing mogroside V based on lipid droplet subcellular organelles and a construction method of the saccharomyces cerevisiae engineering bacteria. The construction method comprises the following steps: integrating squalene epoxidase ERG1, cucurbitadienol synthetase SgCDS, cyclic epoxy hydrolase SgEPH3, cytochrome P450 enzyme CYP87D18 and cytochrome P450 enzyme reductase AtCPR1 at a site GAL80 of a saccharomyces cerevisiae genome; the saccharomyces cerevisiae strain is characterized in that phosphoglucose mutase PGM1, alpha-phosphoglucose mutase PGM2, uridine diphosphate glucose pyrophosphorylase UGP1, glycosyl transferase UGTMG1 and glycosyl transferase SgUGT94-289-3 are integrated at a site Exg1 of a saccharomyces cerevisiae genome. The Saccharomyces cerevisiae engineering bacterium constructed by the invention realizes that the yield of MG-V synthesized by shake flask fermentation is 31.4 mg / L. The Saccharomyces cerevisiae engineering bacterium has the characteristics of strong metabolic flux directionality, high yield and wide industrial application prospect.

This disclosure provides methods for producing a sample of subcellular organelles, particularly nuclei, from a tissue. In some embodiments, this disclosure provides a method of processing a tissue sample involves performing enzymatic / chemical disruption of tissue in a chamber to produce disrupted tissue comprising released cells and / or nuclei and debris; separating the released cells and / or nuclei from the debris therein; and moving the released cells and / or nuclei. In some instances, the method comprises mechanical disruption of the tissue sample.

A specific super-resolution fluorescent probe for lysine-specific demethylase 1 and its application relate to the field of molecular probes. The present invention designs and synthesizes a specific small molecule fluorescent probe LP1 for lysine-specific demethylase 1. The probe is designed based on the active cavity configuration of lysine-specific demethylase 1 and the molecular structure of its specific inhibitor, and has excellent light stability, pH stability, and anti-interference ability against intracellular ions and amino acids. The product released after the probe LP1 reacts with lysine-specific demethylase 1 has an excellent near-infrared fluorescence signal. The probe LP1 successfully and specifically detects the activity of lysine-specific demethylase 1 in different biological samples (pure protein, cell lysate, living cells) and detects the subcellular organelle localization of lysine-specific demethylase 1.

The invention belongs to the field of fluorescent probes, and discloses a fluorescent probe for monitoring dynamic change of hydrogen peroxide in different subcellular organelles as well as a preparation method and application of the fluorescent probe, the probe takes flavonol as a mother nucleus, a hydrogen peroxide ratio type fluorescent probe PML targeting lysosome and a hydrogen peroxide ratio type fluorescent probe PZD targeting lipid droplets. The invention also provides a preparation method and application of the probe. The probe provided by the invention has the advantages of novel structure, simple synthesis method, good recognition effect on hydrogen peroxide, especially hydrogen peroxide in subcellular organelles, fast response speed, high sensitivity, reliable examination result, good biocompatibility, subcellular organelle positioning function, and wide application prospect. The probe can be applied to monitoring of hydrogen peroxide in a microenvironment in cells, and has great significance for applying a probe technology to an actual related pathological model to develop an action mechanism of hydrogen peroxide in pathology of the probe, especially in a ferroptosis process.

The invention discloses a subcellular localization method suitable for floating moss, and belongs to the technical field of mutation or genetic engineering and DNA or RNA and vectors related to the genetic engineering. According to the method disclosed by the invention, the tender thallus of the floating moss is used as a material, and a related protein positioning gene is transferred into a floating moss species through a floating moss stable genetic transformation system. A series of subcellular localization genes are obtained through cloning in a floating moss genome for the first time, the genes can assist GFP fusion protein to enter different subcellular organelles, and a marker line (localization strain) is obtained through stable transformation of a floating moss material. The method can be used for verifying a subcellular localization result of a candidate floating moss target gene, and is a simple, convenient and stable floating moss subcellular localization method. The method has a wide application prospect.

Provided herein are methods and systems for processing preserved tissue and / or dissociating tissue to release cells, nuclei and / or other subcellular organelles. Instruments comprise an interface for engaging cartridges through fluidic lines to introduce or remove liquids from a processing chamber of a cartridge. Systems also include reagent subsystems configured to prevent liquid spills and securely engage fittings between reagent containers and fluidic lines. Systems also include real-time feedback on the pressure on a grinding rotor in a processing chamber of a cartridge.

The invention discloses an amphiphilic heptamethine cyanine fluorescent probe as well as a preparation process and application thereof. According to the fluorescent probe, quaternary ammonium salt ions are introduced to an indole ring on one side of a heptamethine cyanine molecule to increase the hydrophilicity of the molecule, and alkyl chains with different lengths are introduced to an indole ring on the other side of the heptamethine cyanine molecule to regulate and control the hydrophobicity of the molecule. The self-assembled nano particle size of the fluorescent probe molecule is gradually increased by increasing the hydrophobicity, and cell mitochondria targeting is converted into cell membrane targeting. Meanwhile, the change of the self-assembled particle size enhances the tumor targeting ability in a living body, and provides important support for enhancing the curative effect and realizing accurate subcellular organelle targeted tumor phototherapy.

The present invention discloses a 3D cell culture method for ultra-high resolution sub-organelle imaging research and its application, which comprises the following steps: Step (1): Drop the collagen I dilution solution onto the surface of the glass substrate for pre-incubation to form a pre-incubation layer; Step (2): Lay the collagen I and culture medium mixture on the surface of the pre-incubation layer for incubation to enable the collagen I to gel and form a network layer as the sedimentation layer; Step (3): Add the collagen I and cell mixture to the surface of the sedimentation layer for incubation to enable the collagen I to gel; Add cell culture medium and place it in an incubator for 5-7 days to allow the cells to grow and spread, complete the 3D cell culture, and form a 3D cell network layer. Through ultra-high resolution sub-organelle imaging, the sub-organelle structure can be presented, and the intracellular endocytosis kinetics of nanoparticles can be characterized and analyzed in real time for a long time.

The application belongs to the technical field of biological diagnosis, and relates to a subcellular organelle active targeting imaging probe and a preparation method thereof. The subcellular organelle active targeting imaging probe of the application is formed into nanoparticles (NPs) by coupling a fluorescent imaging probe with a special sequence of bombesin peptide and a derivative thereof; on one hand, the NPs can be surface-functionalized with lipophilic cations related to the NPs; on the other hand, the NPs can be passively targeted to tumor sites, and have high permeability and retention effect for solid tumors.

The invention discloses a drug effect evaluation method based on living cell in-situ subcellular organelles and FRET (Fluorescence Resonance Energy Transfer) characterization values. The method comprises the following steps: culturing a cell sample and a control group; selecting different visual fields to carry out E-FRET three-channel imaging and subcellular organelle imaging; performing single cell segmentation to obtain a mask; performing feature extraction by using the mask and the imaged image to obtain FRET efficiency feature information and subcellular organelle feature information of the single cell; calculating an FRET efficiency characterization value and a subcellular organelle characterization value; and performing drug effect evaluation. According to the method, the drug efficacy evaluation is performed by combining the subcellular organelle phenotype characterization value and the FRET efficiency characterization value of the in-vitro living cells after the drug action through in-situ real-time measurement of the cells for the first time, the subcellular organelle phenotype change of the drug reaction is quantified by extracting the characteristics from the subcellular organelle fluorescence image, and the drug efficacy is evaluated from the multi-dimensional characteristics; the dynamic process of the medicine in the cells is comprehensively reflected, the limitation of single index evaluation is avoided, and the accuracy rate is relatively high.

The invention discloses a polypeptide drug chimera and a preparation method thereof, and belongs to the technical field of polypeptide drug carriers. The polypeptide drug chimera is constructed by a recognition segment specifically combined with a target protein and a functional polypeptide carrier mediating the target protein to be transferred to lysosome; an antibody is combined with a lysosome signal amino acid sequence, and pathogenic antigen protein is transferred into a lysosome subcellular organelle for degradation through specific combination of the antibody and different antigens.

Disclosed in the present invention is an ultra-stable red fluorescent protein. The fluorescent protein disclosed in the present invention is a protein having an amino acid sequence of SEQ ID NO: 4. Experiments have demonstrated that the fluorescent protein has great tolerance to electron microscopy sample preparation, can retain more fluorescence signals after an osmium tetroxide treatment and can tolerate embedding in an Epon resin. Moreover, the fluorescent protein has great thermal stability and can retain more fluorescence signals even at 90°C. In addition, the fluorescent protein has relatively high chemical stability and light stability. On the basis of related properties of the fluorescent protein, the protein can be used in the fields of protein labeling, various types of fluorescence imaging, and super-resolution correlative light and electron microscopy, and used for tracking changes in structures and morphologies of samples such as proteins, subcellular organelles, local regions of a cell, and entire cells. The fluorescent protein can be used for packaging related viruses or generate transgenic animals, and for labeling specific proteins, specific organelles or specific cells, thereby achieving rapid tissue clearing imaging and expansion super-resolution microscopy.

The invention provides a chemical gating artificial nano channel based on DNAzyme and application, and belongs to the technical field of nano biotechnology and biosensing. The inner surface of a glass nanotube is modified with a multifunctional DNAzyme component, the outer surface of the glass nanotube is modified with a mitochondrial targeting peptide, targeted capture of a single mitochondrial in a single living cell is achieved, in-situ response to three key molecular signals of Ca < 2 + >, ROS and H < + > in the mitochondrial is achieved by integrating Ca < 2 + > specific DNAzyme and pH / ROS sensitive G4-DNAzyme, and the mitochondrial targeting peptide is used for targeting capture of the mitochondrial in the living cell. Different signal input sequences can generate at least six distinguishable ICR evolution paths, and the action sequence of complex signals of a biological system is dynamically monitored. By innovatively combining DNAzyme molecular recognition, nanochannel confinement effect and subcellular targeting technology, a brand new living cell subcellular organelle monitoring platform is established, and the method has important application value in the fields of cell biology research and clinical diagnosis.

The invention discloses a naphthalimide skeleton pyridinium salt two-photon fluorescent material and application thereof. The structural general formula of the naphthalimide skeleton pyridinium salt two-photon fluorescent material is shown in the specification. The naphthalimide skeleton pyridine salt two-photon fluorescent material disclosed by the invention has a function of developing a plurality of subcellular organelles of cancer cells. Compared with a single-photon fluorescent material, the fluorescent material has the characteristics of low excitation energy, long wavelength, strong penetrability, small light damage, low toxicity and the like, so that the fluorescent material has no damage to normal cells, can be used for cancer cell detection, and has obvious application value.

The invention relates to a method for detecting the state of a DNA nanostructure based on co-localization of a single dye label and a subcellular organelle, and the detection method comprises the following steps: incubating a DNA nanostructure connected with a dye and cells, and detecting the co-localization coefficient of the dye and the cellular organelle; when the co-localization coefficient of the dye and the lysosome is greater than 0.5, judging that the DNA nanostructure is complete; when the co-localization coefficient of the dye and the mitochondria is greater than 0.5, judging that the DNA nano structure is damaged; and judging that the DNA nanostructure is partially complete under other conditions. According to the method for judging the fate of the DNA nanostructure and detecting the integrity of the DNA nanostructure based on co-localization of the single dye and the subcellular organelle, not only can the intact or damaged states of the same DNA nanostructure under different conditions be distinguished by utilizing the position of the DNA nanostructure in the subcellular organelle in the cell, but also the intracellular states of different DNA nanostructures can be detected.

The invention relates to an electron donor (alpha-trithiophene) modified rhodamine dye as well as a synthesis method and application thereof. The dye has strong absorption of 500-600 nm, can efficiently emit in a wave band of 550-700 nm, and is suitable for deep tissue imaging. The structure endows mitochondria with targeting ability, and the probe can be used as a biomarker probe to realize precise positioning of subcellular organelles; meanwhile, active oxygen is efficiently generated through an I-type mechanism under illumination, and the compound is suitable for photodynamic regulation and biosensing for non-disease diagnosis and treatment purposes. In addition, the dye can also be used as an organic photocatalyst for environment or energy related photocatalytic reaction. The invention provides a new choice for a multi-scene photoelectric functional material.

The invention discloses saccharomyces cerevisiae engineering bacteria for efficiently synthesizing mogroside V based on peroxisome subcellular organelles and a construction method of the saccharomyces cerevisiae engineering bacteria. A construction method of the engineering bacterium comprises the following steps: (1) integrating mogrol metabolic enzyme and leucine selection marker Leu2 at a site of a transcription inhibition factor GAL80 of saccharomyces cerevisiae; and (2) integrating peroxisome membrane protein Pex11, carnitine acetyltransferase Cat2, phosphoglucose mutase PGM1, alpha-phosphoglucose mutase PGM2, uridine diphosphate glucose pyrophosphorylase UGP1, glycosyl transferase UGTMG1, glycosyl transferase SgUGT94-289-3 and a histidine selection marker His3 at the site of the saccharomyces cerevisiae glucoside hydrolase Exg1. The engineering strain has the advantages of high yield, strong metabolic flux directionality, wide industrial application prospect and the like, and a new technical route is provided for microbial manufacturing of natural sweeteners.

The invention discloses a whole genetic coding NAD < + > probe, a DNA fragment, an expression vector, a cell and application, and relates to the technical field of biological probes. The invention provides a whole genetic coding NAD < + > probe. The amino acid sequence of the whole genetic coding NAD < + > probe is any one sequence shown in SEQ ID NO: 1 to SEQ ID NO: 9. The invention provides nine NAD < + > probes based on fluorescence resonance energy transfer, which can be used for detecting the concentration of NAD < + >, have high temporal-spatial resolution, high specificity, physiological concentration pH interference resistance and NAD < + > analogue interference resistance, and can reflect the real fluctuation of the concentration of NAD < + > in living cells and subcellular organelles thereof. The fluorescence resonance energy transfer-based probe library for detecting the concentration of NAD + is enriched, and more choices are provided for detecting the concentration of NAD +.