Since the 1990s,continuous technical and scientific advances have defied the diffraction limit in microscopy and enabled three-dimensional(3D)super-resolution imaging.An important milestone in this pursuit is the cohe...Since the 1990s,continuous technical and scientific advances have defied the diffraction limit in microscopy and enabled three-dimensional(3D)super-resolution imaging.An important milestone in this pursuit is the coherent utilization of two opposing objectives(4Pi geometry)and its combination with superresolution microscopy.Herein,we review the recent progress in 4Pi nanoscopy,which provides a 3D,non-invasive,diffraction-unlimited,and isotropic resolution in transparent samples.This review includes both the targeted and stochastic switching modalities of 4Pi nanoscopy.The schematics,principles,applications,and future potential of 4Pi nanoscopy are discussed in detail.展开更多
Objective To estimate the predictive value of double fluorescence [acridine orange （AO）/propidium iodide （PI）] staining results for fertilization rate and clinical outcomes and analyze the correlation between th...Objective To estimate the predictive value of double fluorescence [acridine orange （AO）/propidium iodide （PI）] staining results for fertilization rate and clinical outcomes and analyze the correlation between the results of AO/P1 staining and sperm apoptosis. Methods A prospective study was carried out, 235 infertile couples remedied using traditional in vitro fertilization （IVF） were included. Semen collected from 235 patients were stained by fluorescence dye, AO and PL at the same time the spermatozoa apoptosis rate was calculated by using flow cytometry to detect the rate of Annexin V＋/ PI- . The result of fluorescence was divided into 3 groups as green （G）, yellow （Y） and red （R） according to the color of fluorescence. The correlation between the percent of the 3 colors and clinical outcomes and spermatozoa apoptosis rate was evaluated. Results Significant negative correlation was observed between the percentage of Y and fertilization rate （r= -0.42, P=0.04）, no significant correlation was observed between the percentage of G, R and fertilization rate. The percentage of G, Y, R was not significantly different between pregnant patients and non-pregnant patients, respectiw,ly, while the percentage of Y was significant different between miscarriage patients and liveborn patients （r=0.6L P=0.01） and no significant difference exist in the percentage of G and R between liveborn patients and miscarriage patients. There was a significant positive correlation between the percentage of Y and the rate of Annexin V＋/ PI （r=0.53, P=0.04）, and no significant correlation was shown between the percentage of G, R and the rate of Annexin V＋/ PI-.Conclusion The percentage of yellow group of double fluorescence （AO/PI） staining will affect the fertilization rate and the miscarriage rate, and this group of spermatozoa may be connected with the spermatozoa apoptosis.展开更多
Recently,we theoretically demonstrate that utilization of silica nanobeads co-doped with Cy3 and Cy5 molecules instead of single dye molecules asfluorescent labels can enable optical resolutions far beyond the diffrac...Recently,we theoretically demonstrate that utilization of silica nanobeads co-doped with Cy3 and Cy5 molecules instead of single dye molecules asfluorescent labels can enable optical resolutions far beyond the diffraction-limit.Here,we show that by combining the 4Pi microscopy and the novelfluorescent label,it is possible to completely suppress the sidelobes in 4Pi focal spot and significantly enhance the optical resolution in the axial direction.展开更多
基金financially supported by the grants from National Key Research and Development Program of China (2018YFA0701400 and 2018YFE0119000)the Fundamental Research Funds for the Central Universities (2019QNA5006)+2 种基金ZJU-Sunny Photonics Innovation Center (2019-01)Zhejiang Provincial Natural Science Foundation of China (LR18H180001)startup grant from Southern University of Science and Technology
文摘Since the 1990s,continuous technical and scientific advances have defied the diffraction limit in microscopy and enabled three-dimensional(3D)super-resolution imaging.An important milestone in this pursuit is the coherent utilization of two opposing objectives(4Pi geometry)and its combination with superresolution microscopy.Herein,we review the recent progress in 4Pi nanoscopy,which provides a 3D,non-invasive,diffraction-unlimited,and isotropic resolution in transparent samples.This review includes both the targeted and stochastic switching modalities of 4Pi nanoscopy.The schematics,principles,applications,and future potential of 4Pi nanoscopy are discussed in detail.
基金supported by a grant from STCSM (Subject No.:074207002)
文摘Objective To estimate the predictive value of double fluorescence [acridine orange （AO）/propidium iodide （PI）] staining results for fertilization rate and clinical outcomes and analyze the correlation between the results of AO/P1 staining and sperm apoptosis. Methods A prospective study was carried out, 235 infertile couples remedied using traditional in vitro fertilization （IVF） were included. Semen collected from 235 patients were stained by fluorescence dye, AO and PL at the same time the spermatozoa apoptosis rate was calculated by using flow cytometry to detect the rate of Annexin V＋/ PI- . The result of fluorescence was divided into 3 groups as green （G）, yellow （Y） and red （R） according to the color of fluorescence. The correlation between the percent of the 3 colors and clinical outcomes and spermatozoa apoptosis rate was evaluated. Results Significant negative correlation was observed between the percentage of Y and fertilization rate （r= -0.42, P=0.04）, no significant correlation was observed between the percentage of G, R and fertilization rate. The percentage of G, Y, R was not significantly different between pregnant patients and non-pregnant patients, respectiw,ly, while the percentage of Y was significant different between miscarriage patients and liveborn patients （r=0.6L P=0.01） and no significant difference exist in the percentage of G and R between liveborn patients and miscarriage patients. There was a significant positive correlation between the percentage of Y and the rate of Annexin V＋/ PI （r=0.53, P=0.04）, and no significant correlation was shown between the percentage of G, R and the rate of Annexin V＋/ PI-.Conclusion The percentage of yellow group of double fluorescence （AO/PI） staining will affect the fertilization rate and the miscarriage rate, and this group of spermatozoa may be connected with the spermatozoa apoptosis.
基金J.Chen acknowledges thefinancial support from NSFC(Grant No.61078016).
文摘Recently,we theoretically demonstrate that utilization of silica nanobeads co-doped with Cy3 and Cy5 molecules instead of single dye molecules asfluorescent labels can enable optical resolutions far beyond the diffraction-limit.Here,we show that by combining the 4Pi microscopy and the novelfluorescent label,it is possible to completely suppress the sidelobes in 4Pi focal spot and significantly enhance the optical resolution in the axial direction.
