Direct eye visualization of Cy5 fluorescence for immunocytochemistry and in situ hybridization

被引:6
作者
Ferri, GL [1 ]
Isola, J
Berger, P
Giro, G
机构
[1] Univ Cagliari, Dept Cytomorphol, I-09042 Cagliari, Italy
[2] Univ Cagliari, Troina, Italy
[3] Oasi IRCCS, Troina, Italy
[4] Tampere Univ, Inst Med Technol, Tampere, Finland
[5] Univ Hosp, Tampere, Finland
[6] Austrian Acad Sci, Inst Biomed Aging Res, Innsbruck, Austria
[7] CNR, FRAE, I-40126 Bologna, Italy
关键词
Cy5; fluorescence; immunohistochemistry; immunofluorescence; FISH; multiple staining; fluorescence filter; fluorochrome;
D O I
10.1177/002215540004800314
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Cyanine 5.18 (or Cy5) is a fluorochrome emitting in the long-red/far-red range, usually regarded as unsuitable for direct observation by the human eye. We describe here the optimization Bf a direct visualization approach to Cy5 labeling, based on a standard fluorescence microscope with mercury light excitation and applicable to both immunocytochemistry and fluorescent in situ hybridization. Crucial factors were (a) an excitation path in the microscope not absorbing light in the orange-red range, up to 640 nm, (b) a 588-640-nm excitation filter range, distinctly below the excitation optimum for Cy5, (c) a 650-700-nm emission filter range, transmitting the low-wavelength portion of Cy5 emission, and (d) high-efficiency filter set components allowing a narrow gap between excitation and emission ranges without visible cross-talk of excitation light in the emission path.
引用
收藏
页码:437 / 444
页数:8
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