Prometheus Series Instruments; Excitation And Detection For Fluorescence Measurements - Nano Temper Prometheus Series User Manual

1. Prometheus Series Instruments

The Prometheus Series offers nanoDSF technology (see section 2). nanoDSF is the method
of choice for easy, rapid and accurate analysis of protein stability and aggregation, with
applications in protein engineering, membrane protein research, formulation development and
quality control.
The Prometheus Series comprises two instruments. The Prometheus NT.48 is filled with up to
48 single capillaries for most flexible assay design and easy handling. The Prometheus
NT.Plex employs 24-Capillary chips, which enables manual or automated sample loading. It
can be coupled with robotic and liquid handling platforms for unattended operation and
complete automation.
1.1.

Excitation and Detection for Fluorescence Measurements

The Prometheus Series instruments use advanced UV-LEDs for the excitation of fluorophores,
which are superior to UV-lasers in terms of stability, robustness and flexibility. Tryptophan
fluorescence is collected by a proprietary Dual-UV detection unit which is optimized for the
rapid and sensitive acquisition of tryptophan fluorescence at 330 nm and 350 nm.
LED
UV (maximum at 285 nm)
The detection unit scans the sample tray every 3 seconds, recording the fluorescence intensity
at 330 nm and 350 nm (Figure 1). During a thermal unfolding experiment, the samples are
measured continuously, yielding ultra-high resolution with more than 20 data points per minute.
In case the instrument is equipped with the optional Aggregation Optics, scattering information
is collected simultaneously and with equal data point density.
Figure 1: Prometheus high precision capillary format. Capillaries containing 10 µl sample are loaded onto a
thermal element, which can be set to temperatures from 15 °C-95 °C. Within 3 seconds, the whole capillary tray is
read by the Dual-UV detection unit.
Fluorophore
Tryptophan, Tyrosine
Detection
Dual-UV detector:
330 nm and 350 nm
Detection range:
0-20,000 fluorescence units
10