Thermophoretic Trap for Single Amyloid Fibril and Protein Aggregation Studies

Frank Cichos,Michael Mertig,Juliane Adler,Daniel Huster,Juliane Posseckardt,Martin Fränzl,Tobias Thalheim

doi:10.1038/protex.2019.031

Abstract

Thermophoretic Trap for Single Amyloid Fibril and Protein Aggregation Studies

Highlights

The tracking of the dynamics of single molecules and nano-objects over time has attracted growing interest[1,2]
We present a force-free trapping method in this protocol, which is able to confine the motion of these objects to a region of interest
The amyloid fibrils are trapped in the temperature minimum created above the structure since most molecules and nano-objects move towards the cold

Summary

Introduction

The tracking of the dynamics of single molecules and nano-objects over time has attracted growing interest[1,2]. **Fig. 1: Sketch of the trapping mechanism.** A static temperature field is generated by a chrome ring which is heated by a focused laser beam rotating at a high frequency on the ring. This protocol describes the setup for such single-fibril trapping experiments and provides a step-by-step manual for the production of the chrome structure using UV lithography, the preparation of Aβ40, Aβ42 and pGlu3-Aβ3-4017 amyloid fibrils as example systems, and setting up a measurement sample. Spin coater \ (WS-650S-6NPP/Lite Spin Processor with 650 Controller, Laurell Technology Corporation)

Procedure

Anticipated Results