دورية أكاديمية
أعرض في EDS

Simultaneous multiple-excitation multiphoton microscopy yields increased imaging sensitivity and specificity.

التفاصيل البيبلوغرافية
العنوان: Simultaneous multiple-excitation multiphoton microscopy yields increased imaging sensitivity and specificity.
المؤلفون: Butko MT; Department of Neuroscience and Biomedical Graduate Program, Howard Hughes Medical Institute, University of California, San Diego, La Jolla, CA 92093, USA., Drobizhev M, Makarov NS, Rebane A, Brinkman BC, Gleeson JG
المصدر: BMC biotechnology [BMC Biotechnol] 2011 Mar 02; Vol. 11, pp. 20. Date of Electronic Publication: 2011 Mar 02.
نوع المنشور: Evaluation Study; Journal Article; Research Support, N.I.H., Extramural
اللغة: English
بيانات الدورية: Publisher: BioMed Central Country of Publication: England NLM ID: 101088663 Publication Model: Electronic Cited Medium: Internet ISSN: 1472-6750 (Electronic) Linking ISSN: 14726750 NLM ISO Abbreviation: BMC Biotechnol Subsets: MEDLINE
أسماء مطبوعة: Original Publication: [London] : BioMed Central, 2001-
مواضيع طبية MeSH: Microscopy, Fluorescence, Multiphoton/*methods, Animals ; Cell Line ; Cells/metabolism ; Humans ; Sensitivity and Specificity
مستخلص: Background: Multiphoton microscopy (MPM) offers many advantages over conventional wide-field and confocal laser scanning microscopy (CLSM) for imaging biological samples such as 3D resolution of excitation, reduced phototoxicity, and deeper tissue imaging. However, adapting MPM for critical multi-color measurements presents a challenge because of the largely overlapping two-photon absorption (TPA) peaks of common biological fluorophores. Currently, most multi-color MPM relies on the absorbance at one intermediate wavelength of multiple dyes, which introduces problems such as decreased and unequal excitation efficiency across the set of dyes.
Results: Here we describe an MPM system incorporating two, independently controlled sources of two-photon excitation whose wavelengths are adjusted to maximally excite one dye while minimally exciting the other. We report increased signal-to-noise ratios and decreased false positive emission bleed-through using this novel multiple-excitation MPM (ME-MPM) compared to conventional single-excitation MPM (SE-MPM) in a variety of multi-color imaging applications.
Conclusions: Similar to the tremendous gain in popularity of CLSM after the introduction of multi-color imaging, we anticipate that the ME-MPM system will further increase the popularity of MPM. In addition, ME-MPM provides an excellent tool to more rapidly design and optimize pairs of fluorescence probes for multi-color two-photon imaging, such as CFP/YFP or GFP/DsRed for CLSM.
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معلومات مُعتمدة: DK80506 United States DK NIDDK NIH HHS; P30 NS047101 United States NS NINDS NIH HHS
تواريخ الأحداث: Date Created: 20110304 Date Completed: 20110613 Latest Revision: 20211020
رمز التحديث: 20231215
مُعرف محوري في PubMed: PMC3062589
DOI: 10.1186/1472-6750-11-20
PMID: 21366923
قاعدة البيانات: MEDLINE
الوصف
تدمد:1472-6750
DOI:10.1186/1472-6750-11-20