一种无线离心力显微镜（CFM）能够在商用离心机中进行多重单分子实验。

A wireless centrifuge force microscope (CFM) enables multiplexed single-molecule experiments in a commercial centrifuge.

作者信息

Hoang Tony, Patel Dhruv S, Halvorsen Ken

机构信息

Department of Chemistry, University at Albany, State University of New York, 1400 Washington Avenue, Albany, New York 12222, USA.

Department of Biology, University at Albany, State University of New York, 1400 Washington Avenue, Albany, New York 12222, USA.

出版信息

Rev Sci Instrum. 2016 Aug;87(8):083705. doi: 10.1063/1.4961477.

DOI:10.1063/1.4961477

PMID:27587129

Abstract

The centrifuge force microscope (CFM) was recently introduced as a platform for massively parallel single-molecule manipulation and analysis. Here we developed a low-cost and self-contained CFM module that works directly within a commercial centrifuge, greatly improving accessibility and ease of use. Our instrument incorporates research grade video microscopy, a power source, a computer, and wireless transmission capability to simultaneously monitor many individually tethered microspheres. We validated the instrument by performing single-molecule force shearing of short DNA duplexes. For a 7 bp duplex, we observed over 1000 dissociation events due to force dependent shearing from 2 pN to 12 pN with dissociation times in the range of 10-100 s. We extended the measurement to a 10 bp duplex, applying a 12 pN force clamp and directly observing single-molecule dissociation over an 85 min experiment. Our new CFM module facilitates simple and inexpensive experiments that dramatically improve access to single-molecule analysis.

摘要

离心力显微镜（CFM）最近作为一种用于大规模平行单分子操作和分析的平台被引入。在此，我们开发了一种低成本且自成一体的CFM模块，它可直接在商用离心机内工作，极大地提高了可及性和易用性。我们的仪器集成了研究级视频显微镜、电源、计算机以及无线传输功能，能够同时监测许多单独拴系的微球。我们通过对短DNA双链体进行单分子力剪切来验证该仪器。对于一个7碱基对的双链体，我们观察到由于从2皮牛到12皮牛的力依赖性剪切，出现了超过1000次解离事件，解离时间在10 - 100秒范围内。我们将测量扩展到一个10碱基对的双链体，施加12皮牛的力钳，并在一个85分钟的实验中直接观察单分子解离。我们新的CFM模块便于进行简单且低成本的实验，显著改善了单分子分析的可及性。