RK-Spectro Technical Note 53: Determination of your experimental dead time in fluorescence mode

Introduction

This Technical Note demonstrates how to calculate the experimental dead time of your stopped-flow instrument. The described experiment presupposes that the user understands how to design stopped-flow sequences and correctly use Biokine software. The dead time of a stopped-flow refers to the age of the solution when it reaches the observation point; in other words, it is the time for the mixed solution to pass from the centre of the last mixer to the observation point. Besides total flow rate and cuvette volume, the dead time depends on many other different factors. In this technical note, only flow rate and cuvette volume influences will be discussed. A slight difference between the estimated dead time (given by the Biokine software) and the real dead time may be observed. This is due to the hydrodynamics phenomenon….