37 results about "Fluorescent cell" patented technology

The invention belongs to the technical field of biology, and discloses a stable protective agent, a cell antigen dry sheet and a preparation method and application of the dry sheet, the stable protective agent comprises glycerin with the volume fraction being 0.4%, gelatin with the mass volume fraction being 0.1%, EDTA with the mass volume fraction being 0.1 mM, trehalose with the mass volume fraction being 0.03% and resveratrol with the mass volume fraction being 100 [mu] M. The stable protective agent is adopted for preparing the cell antigen dry sheet, and the cell antigen dry sheet is applied to CBA detection. The stable protective agent has good permeability, can penetrate through cell membranes to enter cells, provides a protection effect inside and outside the cells, has moisture retention and oxidation resistance, can prevent cell deformation and cell fixing layer shedding, and can well maintain the stability and reliability of antigen proteins, and the prepared cell antigen dry sheet can be stored for a long time, and has a good application prospect. The antigenicity of a membrane signal index is not damaged. When a positive sample is detected, obvious green fluorescence cells can be seen, red fluorescence quenching is avoided, and co-localization of red fluorescence and green fluorescence can be observed conveniently.

The invention provides a kit for detecting spermatogenic ability of azoospermia patients and application thereof, and belongs to the technical field of reproductive medicines.The kit comprises the following reagents: testis tissue digestive juice and testis single cell flow staining fluid; the testis single cell flow type staining solution comprises a DNA (Deoxyribose Nucleic Acid) staining solution Hoechst 33342 and an RNA (Ribonucleic Acid) staining solution # imgabs0 # RNASelectTM green fluorescent cell staining agent; according to the kit, double staining of DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) is carried out on testis single cells, and a fluorescence activated cell sorting technique is matched, so that refining of testis single cell subgroups of azoospermia patients is realized, and the spermatogenic ability of the azoospermia patients is determined according to a testis single cell subgroup subdivision result. The kit disclosed by the invention is used for pre-operation evaluation of a microscopic sperm extraction operation, is used for judging whether a patient has spermatogenic ability and is suitable for microscopic sperm extraction or not, can also be used for guiding the patient to further clarify the cause of disease, and has important clinical significance.

The invention discloses a fixing reagent and method for double-color fluorescence detection of an anti-NMDAR antibody and application, the fixing reagent comprises paraformaldehyde and a mixture of methanol and acetone, and the mixture comprises 10-25% by volume of methanol, 75-90% by volume of acetone, 10-25% by volume of paraformaldehyde, 10-25% by volume of paraformaldehyde and 10-25% by volume of paraformaldehyde. The fixing effect is better by adopting paraformaldehyde with the mass-volume ratio concentration of 4% and a mixture of methanol with the volume fraction of 15% and acetone with the volume fraction of 85% in two steps. By adopting the fixing liquid and the fixing method, the antigenicity of NMDAR cannot be damaged, obvious green fluorescence cells can be seen in a positive sample, red fluorescence quenching is avoided, and co-localization of red fluorescence and green fluorescence can be observed by using a fluorescence microscope.

The invention relates to the technical field of biological materials, in particular to a biocompatible method for synthesizing quantum dots from living cells, which comprises the following steps: in the living cell culture process, adding an organic selenium compound containing Se-Se or Se-S bonds into a culture medium, and incubating to obtain selenized cells; further culturing by using a culture medium containing a metal ion-glutathione complex to obtain quantum dots containing metal elements and Se elements; according to the method, the generation of an active selenium precursor with cytotoxicity, such as hydrogen selenide, is effectively avoided, and meanwhile, the outbreak of active oxygen species in cells is also avoided, so that high quantum dot synthesis efficiency is shown. In-vitro cell experiment results show that the cells are endowed with a fluorescent function and still maintain good cell viability, and the cell viability is maintained at 80% or above; in-vivo experiment results show that the fluorescent cells with the synthesized quantum dots have good proliferation, tumor formation, invasion and metastasis capabilities in vivo.

A touch display panel having a plurality of light emitting elements; a plurality of color filters formed at positions corresponding to the plurality of light emitting elements; and an infrared fluorescent pattern comprising a plurality of individual infrared fluorescent cell patterns formed at positions corresponding to at least some of the plurality of color filters; and an infrared control film positioned on a light emitting surface of the display panel.

The present invention discloses a fixing agent, method and application for dual-color fluorescence detection of anti-NMDAR antibodies, wherein the fixing agent includes a mixture of paraformaldehyde and methanol and acetone, wherein the methanol volume fraction is 10-25%, the acetone volume fraction is 75-90%, and the fixing effect is better by using a mixture of paraformaldehyde with a mass volume ratio concentration of 4% and a volume fraction of 15% methanol and 85% acetone in two steps. The fixing solution and the fixing method will not damage the antigenicity of NMDAR, and in positive samples, obvious green fluorescent cells can be seen, and there is no red fluorescence quenching, and the co-localization of red fluorescence and green fluorescence can be observed using a fluorescence microscope.

The invention discloses application of a multi-fungus fluorescent staining solution in preparation of a kit for rapidly identifying survival / drug resistance of fungi, and belongs to the technical field of biological detection. The kit can be used for identifying the death and activity of fungi and high-precision rapid drug resistance identification while specific fungi staining is carried out; a fluorescence brightening agent is adopted to mark fungal components in a sample, and the fungal form is clearly observed under a fluorescence microscope. The nucleic acid fluorescent dye capable of selectively permeating and damaging the cell membrane displays the dead and living state of fungi on the basis of fungus dyeing, and the fluorescent dye permeates into cells and is combined with nucleic acid to emit fluorescence. Metabolic reactive dyes of the cells are used for carrying out rapid drug resistance identification on fungi subjected to drug action. The method can simultaneously realize fungal death and activity identification and rapid drug resistance detection, and is simple and convenient to operate, high in detection speed and accurate in result. The kit is suitable for rapid auxiliary diagnosis and treatment of clinical fungal infection. When the multi-fungus fluorescent staining solution is used for preparing the drug sensitivity detection kit, the detection time of the kit is only 1-2 hours.

The invention provides a method for quantitatively analyzing immune response strength promoted by an adjuvant and application of the method in adjuvant screening, and relates to the technical field of vaccine adjuvants. According to the method for quantitatively analyzing the immune response strength promoted by the adjuvant, the immune response level of the to-be-detected adjuvant can be predicted only by taking the control adjuvant as a reference substance and taking the number of KikGR red fluorescent cells 6-120 hours after immunostimulation of the combined antigen as an index. The method has the characteristics of noninvasive and continuous tracing, simplifies the flow and period of adjuvant effect evaluation, improves the screening efficiency, reduces the research and development period, and provides an effective technical means for rapid screening of adjuvants.

The invention provides a dyeing method for removing autofluorescence of a single cell sorting micro-fluidic chip and a chip without autofluorescence. The dyeing method comprises the following steps: completely soaking the single-cell sorting micro-fluidic chip in a dye solution for dyeing until the top layer of the chip is dyed into a non-transparent black top layer, thereby obtaining the single-cell sorting micro-fluidic chip without autofluorescence, the dye in the dye solution is black dye; the single cell sorting micro-fluidic chip at least comprises a photosensitive dry film serving as a runner layer. The staining method provided by the technical scheme of the invention can effectively solve the problem of autofluorescence of a single cell sorting micro-fluidic chip prepared from a traditional photosensitive dry film, so that when the stained chip is applied to fluorescence activated cell sorting, recognition of automatically screened fluorescent cells is facilitated, the operation difficulty of fluorescence activated cell sorting is reduced, and the working efficiency is improved. And accuracy and reliability are improved.

The application discloses a double-color fluorescence detection anti-NMDAR antibody fixing reagent, a method and application thereof, and the fixing reagent comprises a mixture of paraformaldehyde, methanol and acetone, wherein the volume fraction of methanol in the mixture is 10-25%, and the volume fraction of acetone is 75-90%; the fixing effect is better when the fixing is carried out in two steps by using 4% paraformaldehyde in mass-volume concentration and the mixture of 15% methanol and 85% acetone in volume fraction. By using the fixing liquid and the fixing method, the antigenicity of NMDAR is not damaged, obvious green fluorescent cells can be observed in positive samples, red fluorescence is not quenched, and co-localization of red fluorescence and green fluorescence can be observed by using a fluorescence microscope.

The application provides a fluorescence cell multilayer microscopic imaging method and system based on multi-feature fusion, and relates to the technical field of microscopic imaging. The method comprises the following steps: acquiring a multilayer Z-axis image sequence of a fluorescence cell sample at an X-Y scanning point; dividing an optimal focal plane image into blocks, and identifying the definition state through a convolutional neural network classification model; searching for a replacement block in a neighboring Z layer according to the classification result for a fuzzy block; and outputting a full field of view image after local registration and feathering fusion. The application solves the problems of unstable automatic focusing and insufficient single focal plane coverage under weak fluorescence conditions by fusing multi-feature definition evaluation and deep learning block classification, and realizes full field of view clear imaging of a fluorescence sample with uneven thickness.

The application belongs to the technical field of biology, and discloses a stabilizing protective agent, a cell antigen dry sheet, and a preparation method and application of the dry sheet. The stabilizing protective agent comprises 0.4% glycerol in volume fraction, 0.1% gelatin in mass volume fraction, 0.1 mM EDTA, 0.03% trehalose in mass volume fraction, and 100 muM resveratrol. The stabilizing protective agent is used to prepare the cell antigen dry sheet and is applied in CBA detection. The stabilizing protective agent has good permeability, penetrates into the cell interior through the cell membrane, provides protection inside and outside the cell, has moisturizing and antioxidant properties, can prevent cell deformation and cell fixation layer from falling off, well maintains the stability and reliability of antigen protein, and the prepared cell antigen dry sheet can be stored for a long time without damaging the antigenicity of the membrane signal index. When a positive sample is detected, obvious green fluorescent cells can be seen, and red fluorescent quenching does not occur, so that the co-localization of red fluorescence and green fluorescence can be observed.

The purpose of the present invention is to provide an extracellular vesicle, a preparation method thereof, and a typing analysis method. By removing the surface N-sugar of the extracellular vesicles, the uptake rate of the extracellular vesicles by specific cells is increased. This technology uses co-culture of living cells and extracellular vesicles, fluorescent labeling, and flow cytometry to detect fluorescent cells and fluorescence intensity in cell suspensions in different cell culture dishes. The interaction between living cells and extracellular vesicles is used to perform typing analysis and verification of the extracellular vesicles. In addition, the invention provides a method for preparing extracellular vesicles that can be more efficiently taken up by HCT116.

1. Name of the Design Product: Inverted Fluorescence Cell Imager. 2. Use of the Design Product: It is used for an inverted fluorescence cell imager, which is used for real-time fluorescence observation and measurement of living cells in biological and medical research. 3. Design Key Points of the Design Product: Lies in the shape. 4. Picture or Photo that Best Illustrates the Design Key Points: Perspective view.

The invention discloses a head and neck cancer chemotherapy recurrence animal model and a construction method and application thereof, and relates to the technical field of animal model construction. Comprising the following steps: S1, culturing head and neck cancer tumor cells with fluorescence labels under an in-vitro condition; s2, subcutaneously planting the head and neck cancer tumor fluorescent cells in a groin part of a model animal; s3, 5 days after planting, imaging and observing subcutaneous tumor formation of the model animal, and performing periodic chemotherapy; s4, monitoring through a living body imaging system until a tumor fluorescence signal basically disappears, and determining tumor regression; s5, stopping chemotherapy, continuing feeding for 5-6 weeks, and observing the tumor recurrence condition of the model animal to obtain the head and neck cancer chemotherapy recurrence animal model. The method is easy and convenient to operate, short in experimental period and low in consumption, non-invasive dynamic monitoring can be achieved through the living body imaging technology, the variability of the result is small, the success rate is high, and an ideal platform is provided for head and neck cancer recurrence research and drug screening.

Provided is an in-vitro electrophysiological experiment method. The method comprises: injecting into a target tissue or organ an rAAV expressing a low-abundance fluorescent protein, the genome of the rAAV expressing a low-abundance fluorescent protein containing any one among the following a)-c): a) a promoter sequence, a fluorescent protein gene sequence and a PEST sequence; b) a promoter sequence, a fluorescent protein gene sequence and an ecDHFR DD sequence; and c) a TRE promoter sequence and a fluorescent protein gene sequence, the TRE promoter sequence being located at the upstream of the fluorescent protein gene sequence; and when the fluorescent protein has been expressed for a period of time, performing an in-vitro electrophysiological experiment on the target tissue or organ. The rAAV expresses a low-abundance fluorescent protein, the accumulation of fluorescent proteins in cells is reduced, the cytotoxicity is reduced, and the influence on experimental results is relatively small. In addition, low-abundance fluorescent proteins make cells easier to observe and distinguish from other fluorescent cells in the surrounding area in experiments.

A method of identifying a fluorescent cell in a sample using a microfabricated chip having a top surface including a plurality of microwells. At least one cell of a sample is loaded into at least one microwell of the plurality of microwells. The microfabricated chip is incubated to grow a population of cells from the at least one cell in the at least one microwell. Fluorescence exhibited by at least one microwell is detected by analyzing an image of the microfabricated chip, to thereby determine a presence of a fluorescent cell of interest in the sample.

The cell image registration method provided by the embodiment of the present invention includes: inverting the brightness of the background and target of the first bright field cell image to obtain a second bright field cell image, and using the first fluorescent cell image as the second fluorescent cell image; corroding the one with more pixels covered by the target in the second bright field cell image and the second fluorescent cell image to obtain a third bright field cell image and a third fluorescent cell image; and registering the first bright field cell image and the first fluorescent cell image according to the phase correlation method, the third bright field cell image and the third fluorescent cell image. The embodiment of the present invention first makes the directions of the relative brightness of the background and the target in the bright field cell image and the fluorescent cell image the same, and then corrodes one of the bright field cell image and the fluorescent cell image, and then uses the phase correlation method for registration after the two-step preprocessing, so that the corresponding target centroids in the bright field cell image and the fluorescent cell image are aligned, and the registration is more accurate and stable.

The invention discloses a biological coenzyme resistant reduced methylene blue modified photosensitizer as well as a preparation method and application thereof. The photosensitizer is modified by introducing an electron-donating alkyl group into a methylene blue side chain. The preparation method comprises the following steps: treating specific aniline and benet salt in strong acid by using an oxidizing agent to obtain an intermediate, and reacting in an organic solvent under the action of a catalyst and the oxidizing agent to generate a target product. The photosensitizer can maintain continuous and efficient photodynamic activity in a biological environment, it is guaranteed that active oxygen is efficiently generated under illumination due to the biological coenzyme reduction resisting characteristic of the photosensitizer, and the ROS generation performance is remarkably improved. Through modification, the oil-water distribution coefficient is increased, the hydrophilicity is reduced, the fluorescent powder is more easily taken by cells, bright fluorescence can be observed in a short incubation time, and an intracellular fluorescence signal is greatly enhanced. The photosensitizer realizes effective improvement of the cell killing ability, shows excellent tumor inhibition ability in a tumor model, remarkably improves the tumor inhibition rate, and has excellent tumor phototherapy application potential.

The invention discloses application of glycyrrhizic acid in preparation of a medicine for relieving toxic and side effects of osimertinib. In an in-vitro HaCaT cell strain, glycyrrhizic acid can significantly improve the survival rate of keratinocytes under the action of osimertinib, reduce the apoptosis rate, reduce the expression level of Cleved PARP and a DNA damage marker gamma-H2A. X protein of the cells, and reduce the transcriptional level of inflammation-related factors CCL2, CCL27, IL18, TNF alpha and IL6. In an osimertinib-induced mouse skin toxicity model, glycyrrhizic acid can significantly improve skin unhairing and rash symptoms, recover the expression level of Ki67 and improve proliferation disorder caused by osimertinib; the osimertinib can obviously reduce the number of green fluorescent cells in epidermal tissues, recover the thickness of mouse epidermis, improve the phenomenon of epidermis damage, effectively relieve the toxic and side effects of osimertinib, obviously improve rash, unhairing and inflammatory response caused by osimertinib and improve the safety of osimertinib.

A touch display panel having a plurality of light emitting elements; a plurality of color filters formed at positions corresponding to the plurality of light emitting elements; and an infrared fluorescent pattern comprising a plurality of individual infrared fluorescent cell patterns formed at positions corresponding to at least some of the plurality of color filters; and an infrared control film positioned on a light emitting surface of the display panel.

The invention discloses a fluorescence report system for evaluating homologous mediated DNA (deoxyribonucleic acid) repair efficiency in rice cells. According to the reporting system, plasmids for coding CRISPR / Cas and repairing template DNA are delivered into rice cells, so that a nucleotide sequence for coding a fluorescent protein gene is inserted into the tail end of a coding region of a rice histone gene OsH2B (LOCOs01g62230) with high expression quantity at a fixed point through a homologous mediated DNA repairing mechanism, and frame-combined protein coding sequence fusion is formed; according to the method, the cells subjected to precise editing have the property of emitting fluorescence, the proportion of the cells emitting the fluorescence is detected by utilizing flow cytometry (FACS), and the homologous mediated DNA repair efficiency in the rice cells is calculated. The method effectively solves the technical bottlenecks of high cost, long period and complex analysis in evaluation of editing efficiency depending on deep sequencing, can be used for developing a precise gene editing technology based on homologous mediation DNA repair and the like, and has the characteristics of low cost, short period, simplicity and convenience in analysis and the like.

The present disclosure relates to an intelligent event imaging flow cytometer, comprising a microfluidic system, an optical system and a hardware processing system. The microfluidic system is used for continuously importing a cell sample comprising at least two kinds of cells, and exporting each kind of cell according to the classification result fed back by the hardware processing system. The optical system is used for jointly irradiating the cell sample by a bright field light source and a fluorescent light source, and collecting multiple double-channel coupling information simultaneously comprising bright field cells and fluorescent cells. The hardware processing system is used for obtaining the classification result of the target cell according to multiple target coupling information corresponding to the target cell determined in the double-channel coupling information, and feeding back the classification result to the microfluidic system. In the present disclosure, the cell classification in the cell sample can be realized through the information interaction transmission of the three systems, which simplifies the overall structure of the cytometer. At the same time, the cell classification result is improved in accuracy by jointly judging the cell classification through the cell characteristics collected under the bright field and fluorescent light sources.

The invention discloses an immune cell killing activity detection method based on fluorescently labeled tumor target cells, and relates to the technical field of detection.The immune cell killing activity detection method comprises the steps that peripheral blood or umbilical cord blood of a healthy volunteer is collected, mononuclear cells are obtained through separation with a human lymphocyte separation medium and induced into immune effector cells, and multiplication culture is conducted; taking the fluorescently-labeled tumor cells as target cells, and co-incubating the immune effector cells and the target cells for a period of time; collecting a cell product after co-incubation is finished, and detecting the number of fluorescently-labeled positive cells in the co-incubation product by using a flow cytometer or a fluorescent cell counter; and calculating the killing rate of the immune effector cells to the target cells based on the detected number of the positive cells. The scheme is simple and convenient to operate, rapid, accurate, low in cost and free of cell damage.

The invention provides a detection method and a detection device for multicellular clusters in circulation. The detection method specifically comprises the following steps: exciting and receiving fluorescent pulse signals when fluorescent cells flow through a detection area; the exciting light is focused on an interested blood vessel through a microscope objective; when the fluorescently-labeled cells flow through the laser detection part, fluorescence is released; converting into an electric signal through a photoelectric conversion device; the fluorescence pulse signal subsequently enters a signal conversion device; signal conversion is carried out through a comparator; and when the fluorescence trigger signal is transmitted to the imaging device, the lighting equipment is turned on, and the imaging device records an image sequence of the cells flowing through the detection area after receiving the fluorescence trigger signal. According to the detection device and the detection method for the multicellular clusters in circulation, the physiological environment of the human body can be efficiently, quickly and accurately detected, in-vitro operation such as blood sampling and centrifugation is not needed, a detection target does not leave the living body environment, and a detection result can better reflect the real situation in the physiological environment.

The present invention relates to a cell-based method for detecting botulinum toxin or measuring the activity thereof, and relates to: a genetically modified cell into which a gene construct consisting of an anchor protein group and a dim fluorescent peptide group is introduced; and a use thereof, and, more specifically, to: a genetically modified cell into which a gene construct consisting of a cell membrane-binding peptide and a recombinant fluorescent protein coding sequence, which form an anchor protein group, and a botulinum toxin recognition sequence and a dim fluorescence-inducing peptide coding sequence for fluorescent protein expression, which form a dim fluorescence-inducing peptide group, is introduced; and a use thereof. According to the method of the present invention, botulinum toxin can be detected or the activity thereof can be quantitatively analyzed through fluorescence measurement without using cell lines that are difficult to culture, such as stem cells.

The invention discloses a virus protein dynamic monitoring system and method based on a fluorescent gene editing technology, and relates to the technical field of fluorescent tracer, the system comprises a gene editing module, a fluorescent gene and a Tax response promoter are used for constructing a co-transcription unit by using CRISPR / Cas9, the co-transcription unit is inserted into a virus genome non-functional region through homologous recombination repair, and the gene editing module is used for editing the gene. Subculturing and screening a stable cell line with the fluorescence positive rate reaching the standard; the signal response module is used for realizing synchronous expression of fluorescent protein and Tax through a co-transcription mechanism; the cell carrier module simulates infection through co-culture; the detection and analysis module quantifies the fluorescence intensity and the number of fluorescent cells through a fluorescence microscope and a flow cytometer, the detection period is shortened, the detection efficiency is improved, the system does not need antibiotic treatment, and Tax dynamics can be monitored in real time.

The invention discloses a method for detecting the infectivity of oncolytic viruses in urine and swab preserving fluid based on an ELISpot technology, and relates to the technical field of detection of the infectivity of the oncolytic viruses in the urine and swab preserving fluid, and the specific method comprises the following steps: firstly, placing a proper amount of cells in a transparent cell culture plate for overnight culture, then transferring a sample into a cell plate, and carrying out ELISA detection on the cell plate; culturing for a period of time. An indirect immunofluorescence method is established by adopting an immunofluorescence technology and taking a rabbit anti-Vaccinia Virus Polyclonal Antibody polyclonal antibody as a primary antibody and an Alexa FluorR 488 labeled goat anti-rabbit antibody as a secondary antibody, and the virus infectivity is judged according to the quantity of fluorescent cells infected by the virus. The invention fills the technical blank in the field of detecting the infectivity of the oncolytic virus by using the ELISpot technology, is accurate in detection and high in sensitivity, and has important significance for new drug research and development of oncolytic virus drugs.

The invention discloses a cell communication signal acquisition method and system, and belongs to the technical field of three-dimensional cell optical microscopic imaging. The invention aims to solve the problem that a clear image of a cell communication signal cannot be obtained through single photographing due to small axial depth of field of a fluorescence microscope. The method comprises the following steps: emitting an excitation light beam to a dyed fluorescent cell, and determining three focusing positions of the dyed fluorescent cell along the depth direction; after the fluorescence emitted by the dyed fluorescent cells is split, four light beams to be collected are obtained; the first light beam is collected after passing through the micro-lens array to obtain light field fluorescence data; the second to fourth beams are respectively collected to obtain fluorescence images from the first position to the third position; and performing cell communication signal reconstruction based on the light field fluorescence data, the first position fluorescence image, the second position fluorescence image and the third position fluorescence image to obtain a visual three-dimensional cell communication signal. The method is used for acquiring the cell communication signal.