Using Pre-Cleaned Coverslips to Optimize Coverslip Preparation for Single-Molecule Fluorescence Microscopy

Using Pre-Cleaned Coverslips to Optimize Coverslip Preparation for Single-Molecule Fluorescence Microscopy

Single-molecule fluorescence microscopy has become a popular tool for exploring structural changes and dynamics of biological systems. In our laboratory, we use single-molecule techniques to track conformational changes of immobilized nitric oxide synthase (NOS) and determine their relationship to catalytic activity. Properly immobilizing biomolecules like NOS on a glass surface requires careful attention to coverslip cleaning and preparation. There are many protocols available for cleaning glass coverslips, but these protocols are time-consuming and often use harsh conditions. Alternatively, commercially cleaned and passivated coverslips are available but are quite expensive. In this poster, we examine the possibility of using purchased pre-cleaned coverslips (Schott Nexterion) that come ready to be prepared for single-molecule measurements. We present figures and measurements of merit comparing the pre-cleaned coverslips with ozone cleaned coverslips; demonstrating the effectiveness of pre-cleaned coverslips for single-molecule fluorescence microscopy.