The LightCycler® 480 System is a compact and versatile benchtop real-time PCR platform that can hold multiwell plates for either 96 or 384 samples.
The LightCycler® 480 System is capable of running PCRs over a very broad range of volumes, ranging from 5 to 20 μl (384-well plate) and 20 to 100 μl (96-well plate). This allows for a highly flexible assay design, for example, when sensitivity is crucial because of low target concentrations. Special multiwell plates to optimally fit the LightCycler®480 thermal block are an integral part of the system. They are used in combination with sealing foils that prevent evaporation and contamination. Bar codes on each plate allow easy sample tracking.
For fluorescence excitation, the LightCycler® 480 Instrument uses a high-intensity xenon lamp. The broad spectrum of this light source supports the use of a wide range of dyes and at the same time provides maximal sensitivity for a broad range of assay formats in all channels.
To make excitation as specific as possible and to reduce crosstalk between channels when more than one dye is used, the LightCycler® 480 Instrument includes filters (five different wavelength maxima – 440, 465, 498, 533 and 618 nm – for excitation; six wavelength maxima – 488, 510, 580, 610, 640, 660 nm – for detection). These filters can be flexibly combined according to the requirements of the probe format and dyes used in an experiment.
For signal capture, a large field lens and a charge-coupled device (CCD) camera are specifically arranged to attain uniform collection of signals across the plate. Fluorescence detection is therefore largely independent of sample position and can be done in the absence of additional dyes (e.g., ROX) used otherwise to compensate for two dimensional differences in intensity.