Imaging of human meiotic chromosomes by scanning near-field optical microscopy (SNOM)

被引：13
作者：
Hausmann, M
Liebe, B
Perner, B
Jerratsch, M
Greulich, KO
Scherthan, H
机构：
[1] Inst Mol Biotechnol, Dept Single Cell & Single Mol Tech, D-07708 Jena, Germany
[2] Max Planck Inst Mol Genet, D-14195 Berlin, Germany
[3] Univ Kaiserslautern, Dept Human Biol, D-67653 Kaiserslautern, Germany
来源：
MICRON | 2003年 / 34卷 / 08期
关键词：
immuno-fluorescence; scanning near-field optical microscopy; meiotic chromosomes; synaptonemal complex; telomere;
D O I：
10.1016/S0968-4328(03)00021-0
中图分类号：
TH742 [显微镜];
学科分类号：
摘要：
Centromeres and telomeres are key structures of mitotic and meiotic chromosomes. Especially telomeres develop particular structural properties at meiosis. Here, we investigated the feasibility of scanning near-field optical microscopy (SNOM) for light-microscopic imaging of meiotic telomeres in the sub-hundred nanometer resolution regime. SNOM was applied to visualise the synaptonemal complex (SC) and telomere proteins (TRF1, TRF2)after differential immuno-fluorescent labelling. We tested and compared two different preparation protocols for their applicability in a SNOM setting using micro-fabricated silicon nitride aperture tips. Protocol I consisted of differential labelling of meiotic chromosome cores (SC) by SCP3 immuno-fluorescence and telomeres by TRF1 or TRF2 immuno-fluorescence, while protocol II combined absorption labelling with alkaline phosphatase substrates of cores with fluorescent labelling of telomeres. The results obtained indicate that protocol I reveals a better visualisation of structural (topographic) details than protocol II. By means of SNOM, meiotic chromosome cores could be visualised at a resolution overtopping that of far-field light microscopy. (C) 2003 Elsevier Ltd. All rights reserved.
引用
收藏
页码：441 / 447
页数：7
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