Bentolila Laurent A, Weiss Shimon
Department of Chemistry and Biochemistry, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095-1569, USA.
Cell Biochem Biophys. 2006;45(1):59-70. doi: 10.1385/CBB:45:1:59.
We report a rapid method for the direct multicolor imaging of multiple subnuclear genetic sequences using novel quantum dot-based fluorescence in situ hybridization (FISH) probes (QD-FISH). Short DNA oligonucleotides were attached on QDs and used in a single hybridization/detection step of target sites in situ. QD-FISH probes penetrate both intact interphase nuclei and metaphase chromosomes and showed good targeting of dense chromatin domains with minimal steric hindrances. We further demonstrated that QD's broad absorption spectra allowed different colored probes specific for distinct subnuclear genetic sequences to be simultaneously excited with a single excitation wavelength and imaged free of chromatic aberrations in a single exposure. Thus, these results demonstrate that QD-FISH probes are very effective in multicolor FISH applications. This work also documents new possibilities of using QD-FISH probes detection down to the single molecule level.
我们报告了一种使用基于新型量子点的荧光原位杂交（FISH）探针（QD-FISH）对多个亚核基因序列进行直接多色成像的快速方法。短DNA寡核苷酸连接在量子点上，并用于原位靶位点的单一杂交/检测步骤。QD-FISH探针可穿透完整的间期细胞核和中期染色体，并显示出对致密染色质结构域的良好靶向性，且空间位阻最小。我们进一步证明，量子点的宽吸收光谱允许对不同亚核基因序列具有特异性的不同颜色探针在单一激发波长下同时激发，并在单次曝光中无色差成像。因此，这些结果表明QD-FISH探针在多色FISH应用中非常有效。这项工作还记录了使用QD-FISH探针检测单分子水平的新可能性。
