Synergy H4 Hybrid Multi-Mode Microplate Reader
End point, kinetic, spectral scanning, well-area scanning
Monochromator system 1- to 384-well plates, Filter system 1- to 1536-well plates, PCR plates, Compatible with Take3™ Multi-Volume Plate with 2 µL microspots
4°C above ambient to 65°, C + 0.5°C at 37°C
Fluorescence intensity (FI)
As a result of the illumination, the sample emits light (it fluoresces) and a second optical system collects the emitted light, separates it from the excitation light (using a filter or monochromator system), and measures the signal using a light detector.
Monochromators: Fluorescein 2 pM typical (0.2 fmol/well; 384-well plate) (Top); Fluorescein 2.5 pM typical (0.25 fmol/well; 384-well plate) (Bottom)
Filters/mirrors: Fluorescein 1 pM typical (0.1 fmol/well; 384-well plate)
Monochromators: 250 - 850 nm Filters: 200 - 700 nm (850 nm option)
Monochromators: 5 decades; Filters/mirrors: >6 decades
96: 11 seconds; 384: 22 seconds; 1536: 42 seconds
Fluorescence Polarization (FP)
Fluorescence Polarization can be used to measure the binding constants and kinetics of reactions that cause a change in the rotational time of the molecules. If the fluorophore is bound to a small molecule, the rate at which it tumbles can decrease significantly when it is bound tightly to a large protein. If the fluorophore is attached to the larger protein in a binding pair, the difference in polarization between bound and unbound states will be smaller
3 mP at 1 nM fluorescein typical
400 - 700 nm (320 - 850 nm option)
Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET)
HTRF (TR-FRET) combines two techniques FRET (Fluorescence Resonance Energy Transfer) which is homogeneous and TRF (Time Resolved Fluorescence) which is low background. Advantages of TR-FRET: high sensitivity, high throughput, low false-positive, reliable results and easy protocol.
Europium 60 fM typical with filters (6 amol/well in 384-well plate)
Filters: 200 - 700 nm (850 nm option); Monochromators: 250 - 850 nm
Detect the emission of cold body strong light signal by a substance not resulting from heat.
10 amol ATP typical (flash)
300 - 700 nm
Molecules containing π-electrons or non-bonding electrons (n-electrons) can absorb the energy in the form of ultraviolet or visible light to excite these electrons to higher anti-bonding molecular orbitals. The more easily excited the electrons (i.e. lower energy gap between the HOMO and the LUMO), the longer the wavelength of light it can absorb.
230 - 999 nm, 1 nm increment
2 nm (230-285 nm), 4 nm (>285 nm)
0.0001 OD; Monochromator wavelength accuracy: +2 nm; Monochromator wavelength repeatability: +0.2 nm; Reading speed: 96: 11 seconds; 384: 22 seconds
Minimum Sample Amount
2 μL microspots for low volume assays