There is a need for methods that can image chromosomes with genome-wide coverage, as well as greater genomic and optical resolution. We introduce OligoFISSEQ, a suite of three methods that leverage fluorescence in situ sequencing (FISSEQ) of barcoded Oligopaint probes to enable the rapid visualization of many targeted genomic regions. Applying OligoFISSEQ to human diploid fibroblast cells, we show how four rounds of sequencing are sufficient to produce 3D maps of 36 genomic targets across six chromosomes in hundreds to thousands of cells, implying a potential to image thousands of targets in only five to eight rounds of sequencing. We also use OligoFISSEQ to trace chromosomes at finer resolution, following the path of the X chromosome through 46 regions, with separate studies showing compatibility of OligoFISSEQ with immunocytochemistry. Finally, we combined OligoFISSEQ with OligoSTORM, laying the foundation for accelerated single-molecule super-resolution imaging of large swaths of, if not entire, human genomes.

需要能够对具有全基因组覆盖范围以及更高的基因组和光学分辨率的染色体进行成像的方法。我们推出了 OligoFISSEQ，这是一套包含三种方法的方法，利用带条形码的 Oligopaint 探针的荧光原位测序 (FISSEQ) 来实现许多目标基因组区域的快速可视化。将 OligoFISSEQ 应用于人类二倍体成纤维细胞，我们展示了四轮测序如何足以在数百至数千个细胞中的六条染色体上生成 36 个基因组靶标的 3D 图谱，这意味着只需五到八轮测序即可对数千个靶标进行成像。测序。我们还使用 OligoFISSEQ 以更高分辨率追踪染色体，沿着 X 染色体的路径穿过 46 个区域，单独的研究表明 OligoFISSEQ 与免疫细胞化学的兼容性。最后，我们将 OligoFISSEQ 与 OligoSTORM 结合起来，为大片（如果不是整个）人类基因组的加速单分子超分辨率成像奠定了基础。