22 results about "Subcellular organelle" patented technology

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 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 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.

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.

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.

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 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.

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 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 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 +.

This invention discloses a photosensitive nitric oxide donor that targets both lysosomes and mitochondria, its preparation method, and its applications, belonging to the fields of biomedical materials and medicinal chemistry. The nitric oxide donor has a structure as shown in formula DNOD560, which simultaneously includes a morpholino group for lysosome targeting and a rhodamine fluorophore for mitochondrial targeting, and uses an N-nitroso bond as the photosensitive nitric oxide release unit. The donor undergoes fluorescence quenching under physiological conditions and can undergo N-N bond cleavage under specific wavelength (e.g., 365 nm) illumination, quantitatively releasing nitric oxide and simultaneously generating the strongly fluorescent product DLMF560, enabling self-reporting and visualization of the release process. This donor has advantages such as high release efficiency (95% release efficiency within 70 minutes) and good photostability. Crucially, co-localization experiments have confirmed that it can simultaneously target lysosomes and mitochondria in cells, achieving in-situ, photosensitive release of nitric oxide at the subcellular organelle level, and has been successfully applied to live-cell (A549) imaging, providing a novel tool for studying the function of nitric oxide in subcellular organelles and developing precision treatment strategies.

The application provides an X-ray click chemistry probe based on a synchronous light source and a preparation method and application thereof, and comprises the following steps: 1) synthesizing a polydopamine-metal with a group required for a click reaction on a polydopamine surface; 2) adding a modification reagent to a culture medium with cells to modify biomolecules which can be targeted to various subcellular organelles; 3) adding the polydopamine-metal synthesized in step 1) to the culture medium after modification in step 2) until a click reaction is completed; and 4) adding the cells obtained in step 3) to a substrate suitable for synchronous radiation X-ray imaging, and observing imaging under synchronous radiation X-ray, so that biomolecules in the cells have specific absorption peaks under X-ray, and the method is obtained. According to the application, a method for high-specificity recognition and high-resolution imaging of various biomolecules in cells based on click chemistry and synchronous X-ray is provided, and has a good biomedical application prospect.

The application belongs to the field of fluorescent probes, and discloses a kind of fluorescent probe for monitoring dynamic change of hydrogen peroxide in different subcellular organelles, preparation method and application thereof, the probe is with flavonol as parent nucleus, and hydrogen peroxide ratio type fluorescent probe PML is targeted to lysosome, and hydrogen peroxide ratio type fluorescent probe PZD is targeted to lipid droplet.The application also provides the preparation method and application of the probe.The probe provided by the application has novel structure, simple synthesis method, good recognition effect on hydrogen peroxide, especially hydrogen peroxide in subcellular organelles, fast response speed, high sensitivity, reliable inspection result, good biocompatibility, subcellular organelle positioning function, so that they can be used for monitoring hydrogen peroxide in intracellular microenvironment, and it has great significance to apply probe technology to practical related pathological model to develop the mechanism of hydrogen peroxide in its pathology, especially in the process of ferroptosis.