Qiagen QIAcuity 911000, User Manual

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QIAcuity User Manual 06/2022 24
Optical system
The optical system of the QIAcuity is a camera-based fluorescence microscopy system. The
excitation source for the fluorescence dyes is a high-power white LED. This source in combination
with a specific excitation filter is used to illuminate a whole well at a time. The fluorophores in the
single partitions absorb that light and emit light that is being filtered by a detection filter, collected
and imaged through an objective lens on a CMOS-camera chip (see image below for a detailed
overview of the components). Depending on the configuration of the instrument (2plex/5plex), there
are up to 2 or 5 selectable detection channels. An additional channel is used for detecting the base
fluorescence of the master mix, to determine the exact number of filled partitions and normalization
of fluorescence data.
Scheme of the imaging module.
Available channels
Table 2. Available channels in QIAcuity
Channel Excitation (nm) Emission (nm) Example fluorophores
Green 463–503 518–548
FAM™, EvaGreen
®
Yellow 514–535 550–564
HEX™, VIC
®
Orange 543–565 580–606
TAMRA™
Red 570–596 611–653
ROX™, Texas Red
®
Crimson 590–640 654–692
Cy5
®