Bioimaging最新文献_第9页

Two-photon laser scanning fluorescence microscopy-from a fluorophore and specimen perspective 从荧光团和样品的角度看双光子激光扫描荧光显微镜

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<168::AID-BIO7>3.0.CO;2-8

J D Bhawalkar, A Shih, S J Pan, W S Liou, J Swiatkiewicz, B A Reinhardt, P N Prasad, P C Cheng

引用次数: 25

Multiphoton excitation cross‐sections of molecular fluorophores 分子荧光团的多光子激发截面

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<198::AID-BIO10>3.0.CO;2-X

Chris Xu, Rebecca M. Williams, W. Zipfel, W. Webb

引用次数: 153

Effect of the detector size and the fluorescence wavelength on the resolution of three- and two-photon confocal microscopy 探测器尺寸和荧光波长对三光子和双光子共聚焦显微镜分辨率的影响

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<129::AID-BIO3>3.0.CO;2-L

Min Gu, X S Gan

引用次数: 13

Multiphoton excitation of the DNA stains DAPI and Hoechst DNA染色剂DAPI和Hoechst的多光子激发

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<138::AID-BIO4>3.0.CO;2-K

Ignacy Gryczynski, Henryk Malak, Joseph R Lakowicz

{"title":"Multiphoton excitation of the DNA stains DAPI and Hoechst","authors":"Ignacy Gryczynski, Henryk Malak, Joseph R Lakowicz","doi":"10.1002/1361-6374(199609)4:3<138::AID-BIO4>3.0.CO;2-K","DOIUrl":"https://doi.org/10.1002/1361-6374(199609)4:3<138::AID-BIO4>3.0.CO;2-K","url":null,"abstract":"The DNA stains 40′ ,6-diamidino-2-phenylindole hydrochloride (DAPI) and Hoechst 33342 were found to display two- or three-photon excitation from 810 to 910 nm. We examined the effect of excitation wavelength on the mode of excitation for DAPI and Hoechst 33342 in the solvent isobutanol and when bound to double helical DNA. For DAPI and Hoechst 33342 in isobutanol the mode of excitation changed from two- to three-photon excitation over this wavelength range, with apparent transition wavelengths of 855 and 880 nm, respectively. However, when bound to DNA, the transition wavelength from two- to three-photon excitation increased for both probes. In the case of DAPI-DNA, the apparent transition wavelength increased to 868 nm, and three-photon excitation occurred above 900 nm. For Hoechst 33342-DNA the mode of excitation was a mixture of two- and three-photon excitation to the longest excitation wavelength of 910 nm, and we were unable to observe pure three-photon excitation for Hoechst 33342-DNA. In the transition region the anisotropy of DAPI was dependent on laser power, illustrating that the mode of excitation and transition wavelengths will depend on the precise experimental conditions. Higher spatial resolution was observed for three-photon excitation of DAPI than for two-photon excitation of Hoechst 33342. These results suggest that these probes can be used for either two- or three-photon imaging of DNA or chromosomes.","PeriodicalId":100176,"journal":{"name":"Bioimaging","volume":"4 3","pages":"138-148"},"PeriodicalIF":0.0,"publicationDate":"1996-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1361-6374(199609)4:3<138::AID-BIO4>3.0.CO;2-K","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72354780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mechanisms of photobleaching investigated by fluorescence correlation spectroscopy 荧光相关光谱法研究光漂白机理

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<149::AID-BIO5>3.0.CO;2-D

Jerker Widengren, Rudolf Rigler

{"title":"Mechanisms of photobleaching investigated by fluorescence correlation spectroscopy","authors":"Jerker Widengren, Rudolf Rigler","doi":"10.1002/1361-6374(199609)4:3<149::AID-BIO5>3.0.CO;2-D","DOIUrl":"https://doi.org/10.1002/1361-6374(199609)4:3<149::AID-BIO5>3.0.CO;2-D","url":null,"abstract":"Fluorescence correlation spectroscopy (FCS) can be used to investigate the photobleaching properties of fluorophores in solution. The advantage with this method is that in addition to the photobleaching rate the formation and decay rates of the triplet state can be measured. In this way, it is possible to calculate the photodestruction quantum yield and relate the photostability of a fluorescent compound in a certain environment to the photodynamical behaviour of the singlet-triplet transitions. This is likely to contribute to a better understanding of the mechanisms of photobleaching given the central importance of dye triplet states in photobleaching processes. The approach was applied to the measurement and characterization of the photobleaching of Rh6G in aqueous solution and FITC in 1 mM sodium carbonate buffer (pH 9). The photobleaching yields measured are discussed in view of the simultaneous triplet properties at different excitation intensities, oxygen concentrations as well as in the presence or absence of quencher molecules. This study suggests that FCS is likely to provide a valuable tool for the elucidation of the mechanisms of photobleaching, which are far from understood in all their details.","PeriodicalId":100176,"journal":{"name":"Bioimaging","volume":"4 3","pages":"149-157"},"PeriodicalIF":0.0,"publicationDate":"1996-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1361-6374(199609)4:3<149::AID-BIO5>3.0.CO;2-D","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72354782","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 166

Using offset interfering beams for improved resolution in confocal imaging: the potential of the PSAF-technique 利用偏移干涉光束提高共聚焦成像的分辨率:psaf技术的潜力

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<179::AID-BIO8>3.0.CO;2-1

M. Müller, G. Brakenhoff

引用次数: 5

Multiphoton excitation cross-sections of molecular fluorophores 分子荧光团的多光子激发截面

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<198::AID-BIO10>3.0.CO;2-X

Chris Xu, R M Williams, Warren Zipfel, Watt W Webb

{"title":"Multiphoton excitation cross-sections of molecular fluorophores","authors":"Chris Xu, R M Williams, Warren Zipfel, Watt W Webb","doi":"10.1002/1361-6374(199609)4:3<198::AID-BIO10>3.0.CO;2-X","DOIUrl":"https://doi.org/10.1002/1361-6374(199609)4:3<198::AID-BIO10>3.0.CO;2-X","url":null,"abstract":"Nonlinear excitation of fluorophores through molecular absorption of two or three near-infra-red photons from the tightly focused femtosecond pulses of a mode-locked laser offers the cellular biologist an unprecedented panoply of biomolecular indicators for microscopic imaging and cellular analysis. Measurements of the two-photon excitation spectra of 25 ultra-violet and visible absorbing fluorophores from 690 to 1050 nm reveal useful cross sections for near infra-red excitation, providing an artist's palette of emission markers, chemical indicators, and native cellular absorbers for living biological preparations. Measurements of three-photon fluorophore excitation spectra now suggest relatively benign wavelengths to excite deeper UV fluorophores. The inherent optical sectioning capabilities of focused nonlinear excitation provides three-dimensional resolution for imaging and avoids out-of-focus background. Measured nonlinear excitation spectra are described and implications to nonlinear microscopy for biological imaging are defined.","PeriodicalId":100176,"journal":{"name":"Bioimaging","volume":"4 3","pages":"198-207"},"PeriodicalIF":0.0,"publicationDate":"1996-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1361-6374(199609)4:3<198::AID-BIO10>3.0.CO;2-X","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72324630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 156

Image formation in three-photon fluorescence microscopy 三光子荧光显微镜成像

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<124::AID-BIO2>3.0.CO;2-2

C J R Sheppard

引用次数: 16

Mechanisms of photobleaching investigated by fluorescence correlation spectroscopy 荧光相关光谱法研究光漂白机理

Bioimaging Pub Date : 1996-09-01 DOI: 10.1002/1361-6374(199609)4:3<149::AID-BIO5>3.0.CO;2-D

J. Widengren, R. Rigler

{"title":"Mechanisms of photobleaching investigated by fluorescence correlation spectroscopy","authors":"J. Widengren, R. Rigler","doi":"10.1002/1361-6374(199609)4:3<149::AID-BIO5>3.0.CO;2-D","DOIUrl":"https://doi.org/10.1002/1361-6374(199609)4:3<149::AID-BIO5>3.0.CO;2-D","url":null,"abstract":"Fluorescence correlation spectroscopy (FCS) can be used to investigate the photobleaching properties of fluorophores in solution. The advantage with this method is that in addition to the photobleaching rate the formation and decay rates of the triplet state can be measured. In this way, it is possible to calculate the photodestruction quantum yield and relate the photostability of a fluorescent compound in a certain environment to the photodynamical behaviour of the singlet-triplet transitions. This is likely to contribute to a better understanding of the mechanisms of photobleaching given the central importance of dye triplet states in photobleaching processes. The approach was applied to the measurement and characterization of the photobleaching of Rh6G in aqueous solution and FITC in 1 mM sodium carbonate buffer (pH 9). The photobleaching yields measured are discussed in view of the simultaneous triplet properties at different excitation intensities, oxygen concentrations as well as in the presence or absence of quencher molecules. This study suggests that FCS is likely to provide a valuable tool for the elucidation of the mechanisms of photobleaching, which are far from understood in all their details.","PeriodicalId":100176,"journal":{"name":"Bioimaging","volume":"74 1","pages":"149-157"},"PeriodicalIF":0.0,"publicationDate":"1996-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79676853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 163

Fluorescent dot counting in interphase cell nuclei 间期细胞核荧光点计数

Bioimaging Pub Date : 1996-06-01 DOI: 10.1002/1361-6374(199606)4:2<93::AID-BIO7>3.0.CO;2-7

H. Netten, L. Vliet, H. Vrolijk, W. Sloos, H. Tanke, I. Young

{"title":"Fluorescent dot counting in interphase cell nuclei","authors":"H. Netten, L. Vliet, H. Vrolijk, W. Sloos, H. Tanke, I. Young","doi":"10.1002/1361-6374(199606)4:2<93::AID-BIO7>3.0.CO;2-7","DOIUrl":"https://doi.org/10.1002/1361-6374(199606)4:2<93::AID-BIO7>3.0.CO;2-7","url":null,"abstract":"Fluorescence in situ hybridization allows the enumeration of chromosomal abnormalities in interphase cell nuclei. This process is called dot counting. To estimate the distribution of chromosomes per cell, a large number of cells have to be analysed, particularly when the frequency of aberrant cells is low. Automation of dot counting is desirable because manual counting is tedious, fatiguing, and time consuming. We have developed a completely automated fluorescence microscope system that counts fluorescent hybridization dots for one probe in interphase cell nuclei. This system works with two fluorescent dyes—one for the DNA hybridization dots and one for the cell nucleus. A fully automated scanning procedure has been used for the image acquisition. After an image is acquired it has to be analysed in order to find the nuclei and to detect the dots. This article focuses upon the dot detection procedure. Three different algorithms are presented. The problems of 'overlapping' dots and split dots are discussed. The automated dot counter has been tested on a number of normal specimens where DAPI was used for the nucleus counter stain and a centromeric probe was used to mark the chromosome 12. The slides contained lymphocytes from cultured blood. The performance of the different algorithms has been evaluated and compared with manually obtained results. The automated counting results approximate the results of manual counting.","PeriodicalId":100176,"journal":{"name":"Bioimaging","volume":"44 1","pages":"93-106"},"PeriodicalIF":0.0,"publicationDate":"1996-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81011104","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 54