Synergy-2 Multi-mode Microplate Reader: Absorbance,Fluorescence & Luminescence

The Synergy-2 Multimode Microplate Reader

The BioTek Synergy 2 offers three modes of operation for 6- to 384-well plates:

  1. Absorbance,

  2. Luminescence,

  3. Fluorescence.

Absorbance can be measured in the UV and Visible ranges, up to 4.0 OD, with a resolution of 0.001 OD (absorbance units). Any wavelength between 200 and 999 nm can be selected with 1 nm increments. A Xenon flash is used as light source and a monochromator serves for wavelength selection. Three basic modes of absorbance measurement can be selected: 1. Concentration measurements using calibration curves, 2. Kinetic measurements carried out by recording absorbance changes versus time and 3. Absorbance Spectra that are obtained by measuring absorbance as function of wavelength. The microplate reader is set up for temperature control and for shaking. A reading of a 96-well plate can be performed in as little as 11 seconds.

The Two reagent dispensers

Luminescence between 300 and 700nm can be monitored as a function of time. The system can detect as little as 0.01 femtomole of ATP, and up to six orders of magnitude higher concentrations.

The instrument is equipped with two Reagent Dispensers. The dispensers can be programmed to deliver two different reagents during the reaction time course. A syringe (#1) draws the reagent solution from the vial (#2), and then drives it through the switching valve (#3) towards the wellplate (#4).

The fluorescence system has a tungsten-halogen lamp as light source, and uses filters and dichroic mirrors to select wavelengths for excitation and emission. Synergy-2 can detect 1pM of fluorescein, i.e. 0.2 femtomole / well in a 96-well plate. The dynamic range is > 6 decades. The excitation and emission pathways are each equipped with a four-position filter wheel - the filters are replaceable (see the scheme at the bottom of this page).

Table I shows our current collection of filters. If you plan to use the Synergy-2 for fluorescence measurements, you should first determine what wavelengths you want to use for the excitation and the emission, and then select the appropriate filters from table I. Call Israel Meir at 08-9489810 to set up the instrument for your experiment.

Table I - Optical Filters available for Fluorescence Measurements

wavelength (nm)bandwidth (nm)colorMain Applications
28410uvTryptophan excitation
34030uvNADH excitation and tryptophan emission
36040uvMUB, caspace-3, europium chelate excitation
38020 ImageFura-2 and EBFP excitation
42050 ImageCFP excitation and Quanta-Blu emission
44040 ImageNADH emission
460 40 ImageNanoOrange excitation and EBFP and MUB emission
48520 ImageFluorescein, EGFP excitation and CFP emission
50027 ImageYFP excitation
50820 ImageFura-2 emission
52820 ImageVIC excitation and Fluorescein and EGFP emission
54025 Image Alexa Fluor 546, CY3, and rhod2 excitation and EYFP emission
57515 Image ROX excitation and CY3 and 5-Tamra emission
590 20 ImageAlexa Fluor 594 and Texas Red excitation and Cell Titer Blue emission
60040 Image Alamar Blu, Amplex Red, RFP and porphyrin emission
64540 ImageTexas Red and Propidium iodide emission

Synergy 2 - Optical Scheme of Fluorescence Measurements

Fluorescenc measurement using filters