[ Objective] The paper aimed to search new identification methods of Encephalitozoon cuniculi on tissue sections. [ Method] Using improved Gram staining method and methyl green pyronin staining method, the pathologica...[ Objective] The paper aimed to search new identification methods of Encephalitozoon cuniculi on tissue sections. [ Method] Using improved Gram staining method and methyl green pyronin staining method, the pathological sections of sick rabbits were stained and identified. [ Result] The pathological changes in brain tissue could be clearly observed on sections, but parasites were not examined in pathological brain tissues stained by common staining method. When the pathological section was stained by improved Gram staining method, the pathological changes in brain tissue were not ouly stained very clearly, but blue parasites were also found in brain tissues. The parasites in epithelioid cells were stained into purple ones by methyl green pyronin staining method. [ Conclusion] The im- proved Gram staining method and methyl green pyronin staining method performed good staining effects of E. cuniculi in pathological sections, which were conducive to rapid diagnosis of encephalitozoonosis in rabbit.展开更多
Fluorescence nanoscopy has become increasingly powerful for biomedical research,but it has historically afforded a small field-ofview(FOV)of around 50μm×50μm at once and more recently up to∼200μm×200μm....Fluorescence nanoscopy has become increasingly powerful for biomedical research,but it has historically afforded a small field-ofview(FOV)of around 50μm×50μm at once and more recently up to∼200μm×200μm.Efforts to further increase the FOV in fluorescence nanoscopy have thus far relied on the use of fabricated waveguide substrates,adding cost and sample constraints to the applications.Here we report PRism-Illumination and Microfluidics-Enhanced DNA-PAINT(PRIME-PAINT)for multiplexed fluorescence nanoscopy across millimeter-scale FOVs.Built upon the well-established prism-type total internal reflection microscopy,PRIME-PAINT achieves robust singlemolecule localization with up to∼520μm×520μm single FOVs and 25−40 nm lateral resolutions.Through stitching,nanoscopic imaging over mm^(2)sample areas can be completed in as little as 40 min per target.An on-stage microfluidics chamber facilitates probe exchange for multiplexing and enhances image quality,particularly for formalin-fixed paraffin-embedded(FFPE)tissue sections.We demonstrate the utility of PRIME-PAINT by analyzing∼106 caveolae structures in∼1,000 cells and imaging entire pancreatic cancer lesions from patient tissue biopsies.By imaging from nanometers to millimeters with multiplexity and broad sample compatibility,PRIMEPAINT will be useful for building multiscale,Google-Earth-like views of biological systems.展开更多
Fluorescence labeling and imaging provide an opportunity to observe the structure of biological tissues,playing a crucial role in the field of histopathology.However,when labeling and imaging biological tissues,there ...Fluorescence labeling and imaging provide an opportunity to observe the structure of biological tissues,playing a crucial role in the field of histopathology.However,when labeling and imaging biological tissues,there are still some challenges,e.g.,time-consuming tissue preparation steps,expensive reagents,and signal bias due to photobleaching.To overcome these limitations,we present a deep-learning-based method for fluorescence translation of tissue sections,which is achieved by conditional generative adversarial network(cGAN).Experimental results from mouse kidney tissues demonstrate that the proposed method can predict the other types of fluorescence images from one raw fluorescence image,and implement the virtual multi-label fluorescent staining by merging the generated different fluorescence images as well.Moreover,this proposed method can also effectively reduce the time-consuming and laborious preparation in imaging processes,and further saves the cost and time.展开更多
基金Supported by National Natural Science Foundation of China(31372407)
文摘[ Objective] The paper aimed to search new identification methods of Encephalitozoon cuniculi on tissue sections. [ Method] Using improved Gram staining method and methyl green pyronin staining method, the pathological sections of sick rabbits were stained and identified. [ Result] The pathological changes in brain tissue could be clearly observed on sections, but parasites were not examined in pathological brain tissues stained by common staining method. When the pathological section was stained by improved Gram staining method, the pathological changes in brain tissue were not ouly stained very clearly, but blue parasites were also found in brain tissues. The parasites in epithelioid cells were stained into purple ones by methyl green pyronin staining method. [ Conclusion] The im- proved Gram staining method and methyl green pyronin staining method performed good staining effects of E. cuniculi in pathological sections, which were conducive to rapid diagnosis of encephalitozoonosis in rabbit.
基金supported by the Cancer Early Detection Advanced Research(CEDAR)Center of the OHSU Knight Cancer Institutesupported in part by the OHSU Knight Cancer Institute,the Damon Runyon Cancer Research Foundation,the Cancer Systems Biology Consortium from the National Cancer Institute(CSBC,grant number U54 CA209988,PI:Joe W.Gray)the National Institute of General Medical Sciences(grant number R01 GM132322,PI:X.N.).
文摘Fluorescence nanoscopy has become increasingly powerful for biomedical research,but it has historically afforded a small field-ofview(FOV)of around 50μm×50μm at once and more recently up to∼200μm×200μm.Efforts to further increase the FOV in fluorescence nanoscopy have thus far relied on the use of fabricated waveguide substrates,adding cost and sample constraints to the applications.Here we report PRism-Illumination and Microfluidics-Enhanced DNA-PAINT(PRIME-PAINT)for multiplexed fluorescence nanoscopy across millimeter-scale FOVs.Built upon the well-established prism-type total internal reflection microscopy,PRIME-PAINT achieves robust singlemolecule localization with up to∼520μm×520μm single FOVs and 25−40 nm lateral resolutions.Through stitching,nanoscopic imaging over mm^(2)sample areas can be completed in as little as 40 min per target.An on-stage microfluidics chamber facilitates probe exchange for multiplexing and enhances image quality,particularly for formalin-fixed paraffin-embedded(FFPE)tissue sections.We demonstrate the utility of PRIME-PAINT by analyzing∼106 caveolae structures in∼1,000 cells and imaging entire pancreatic cancer lesions from patient tissue biopsies.By imaging from nanometers to millimeters with multiplexity and broad sample compatibility,PRIMEPAINT will be useful for building multiscale,Google-Earth-like views of biological systems.
基金This work was supported in part by the National Natural Science Foundation of China(61871263,12274092,and 12034005)in part by the Explorer Program of Shanghai(21TS1400200)+1 种基金in part by the Natural Science Foundation of Shanghai(21ZR1405200)in part by the Medical Engineering Fund of Fudan University(YG2022-6).
文摘Fluorescence labeling and imaging provide an opportunity to observe the structure of biological tissues,playing a crucial role in the field of histopathology.However,when labeling and imaging biological tissues,there are still some challenges,e.g.,time-consuming tissue preparation steps,expensive reagents,and signal bias due to photobleaching.To overcome these limitations,we present a deep-learning-based method for fluorescence translation of tissue sections,which is achieved by conditional generative adversarial network(cGAN).Experimental results from mouse kidney tissues demonstrate that the proposed method can predict the other types of fluorescence images from one raw fluorescence image,and implement the virtual multi-label fluorescent staining by merging the generated different fluorescence images as well.Moreover,this proposed method can also effectively reduce the time-consuming and laborious preparation in imaging processes,and further saves the cost and time.
