公开(公告)号：WO2018026873A1

公开(公告)日：2018-02-08

申请号：PCT/US2017/044994

申请日：2017-08-01

Applicant: CALIFORNIA INSTITUTE OF TECHNOLOGY

Inventor： CAI, Long ,  ENG, Chee, Huat ,  SHAH, Sheel

IPC: C12Q1/68 ,  G01N21/64

CPC classification number: C12Q1/6841 ,  G01N21/64 ,  C12Q2525/161 ,  C12Q2563/107 ,  C12Q2563/179 ,  C12Q2565/514

Abstract: The present invention, among other things, provides technologies for detecting and/or quantifying nucleic acids in cells, tissues, organs or organisms. Pre-designed barcodes are associated specific molecular targets through sequential hybridization experiments. A pseudo¬ color based barcoding scheme is developed to overcome limitations in the previous generation of the technology such as lack of visual signals that can be associated with the probes or small internal within cell when carrying out in situ experiments.. The current method can be applied to both in vitro and in situ analysis. According to the method, each barcoding round comprises multiple serial hybridizations where a small number of colored signals (that are associated with probes) are used in each hybridization experiment within a serial hybridization round. Images from each serial hybridization experiment within the same serial hybridization round are combined to form a composite image for each barcoding round. In each barcoding round, the same set of molecular targets are analyzed. After all barcoding rounds are completed, associated of the barcode with these molecular targets is completed.

Abstract translation: 本发明尤其提供了用于检测和/或定量细胞，组织，器官或生物体中的核酸的技术。 预先设计的条形码通过顺序杂交实验与特定的分子靶标相关联。 伪＆不; 开发基于颜色的条形码方案以克服上一代技术中的限制，例如在进行原位实验时缺乏可与探针相关联的视觉信号或细胞内小内部。目前的方法可应用于 体外和原位分析。 根据该方法，每个条形码包含多个连续杂交，其中在连续杂交轮次内的每个杂交实验中使用少量有色信号（与探针相关联）。 来自每个序列杂交实验中的图像在相同的连续杂交轮内被组合以形成每个条形码的合成图像。 在每个条形码中，分析同一组分子靶标。 在所有条形码完成后，条形码与这些分子目标的关联就完成了。