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BMG Labtech Application Notes
Five Measurement Principles Including Full Absorbance Spectrum in One Reader
The new Omega series of microplate readers is the world's first family of multifunctional readers capable of capturing a full absorbance spectrum (220 to 1000 nm) at a resolution of 1 nm. Read times faster than 1 second per well are possible.
POLARstar Omega, the multi-detection microplate reader for life science laboratories, offers the following five detection modes:
- Fluorescence Polarization (FP) - including Far Red™
- Fluorescence Intensity - including FRET
- Time-Resolved Fluorescence (TRF) - including DELFIA™
- High-Performance Luminescence (flash and glow) - including BRET
- UV/Vis Absorbance - measurement of complete spectra or individual wavelengths
The POLARstar Omega has all of the features of the FLUOstar Omega, plus fluorescence polarization and simultaneous dual emission detection. The POLARstar has two emission photomultiplier tubes (PMTs), giving it a unique advantage over other readers in that it can read two emission wavelengths simultaneously. This distinct feature allows the POLARstar to measure up to 50 points per second on both channels.
Because the POLARstar Omega does not have to switch filters or polarizers to read the second channel, its Simultaneous Dual Emission Detection (SDED) makes FP, FRET, and BRET assays faster and more precise.
Single channel FP readers must read the microplate twice - once for parallel emission and once for perpendicular emission. But the POLARstar Omega collects both emission vectors simultaneously for a fantastic speed advantage. The same is true for FRET and BRET experiments that have two or more emission wavelengths.
- Ca2+ measurements (e.g. Ca2+flux with Fura-2, Indo-1, FLUO-3/4)
- Enzyme activity
- FRET/BRET assays
- Cell toxicity, proliferation and viability
- Protein and nucleic acid quantitation (e.g. Bradford, Lowry, DNA, 260/280 nm)
- ATP quantification
- Immunoassays, ELISA
- Luciferase, dual luciferase, BRET with simultaneous dual emission detection
- ORAC and ABEL antioxidant assays
- Fluorescent proteins, including FRETs (e.g. CFP/YFP)
By integrating a high intensity xenon flash lamp with sophisticated light guide technology the POLARstar Omega covers the full wavelength range from 230 to 900 nm for both excitation and emission. The POLARstar Omega can read all plate formats from 6- up to 384-wells in all five detection modes.
The software´s automatic optic switching (AOS) feature allows for controlled changes between detection mode and for switching from top to bottom reading. The "QuickStart" mode allows users to read plates within just 3 mouse clicks. Impressive well-scanning software allows cell-based assays to be performed with ease.
Stable and uniform incubation is achieved by temperature-regulated heating plates above and below the entire microplate movement area. The top incubation plate is at a slightly higher temperature than the bottom plate to help reduce condensation when lids or sealers are used. Incubation up to 60°C is available for users with elevated temperature needs.
Most recently the atmospheric control unit (ACU) was introduced which offers a unique independent control of O2 and Co2. It can be ordered as an option or as an upgrade for excisting Omage users
On-board reagent injectors
The POLARstar Omega can be equipped with one or two integrated syringe injectors (no bulky add-on boxes beside or on top of the microplate reader). The injectors can deliver agonists, substrates, enzymes - even cells, for advanced microplate assays. Injection speed and timing is fully adjustable within the control software. Up to four different volumes, between 5 to 350 µL in 0.5 µL increments, can be delivered to each well; thereby allowing the user to produce dilution schemes, concentration ranges, etc.
Endpoint, slow and fast kinetic measurements
Kinetic data can be collected as fast as 50 reading points per second or as slow as one reading point every 2½ hours. You can capture a fast calcium signal that happens in a few seconds or measure bacterial growth over a period of days. Furthermore, data can be collected at different rates within the same experiment; this allows the user to collect more data points at regions of interest (e.g. growth phase) and less data points in areas of little change (e.g. lag phase).
The POLARstar Omega can be used to measure up to eight fluorophores / lumiphores / chromophores per well. In absorbance mode, the reader simultaneously provides chromatic values at each nanometer from 220 to 1000 nm. This allows the user to perform 260/280 nm DNA quantitation, SNPs, and other multichromatic assays easily. In dual excitation (e.g. FURA-2) and in dual emission (FRET and BRET assays) applications, fast filter switching enables rapid measurement at both wavelengths.
Stacker and robot compatibility
The standardized reader footprint and robotic software interface make it easy to integrate the POLARstar Omega into all existing robotic systems. The Stacker II with a 50 plate capacity and optional barcode reader, is the modular attachment that can be integrated with the POLARstar Omega for medium throughput plate handling.
The Omega Windows™ based software has an easy to use control and a complete new powerful data evaluation package. It is compatible with Windows 7/ XP/ 2000 and Vista™ operating system and is fully compliant with FDA regulation 21 CFR Part 11.
The control software allows flexible definition of microplate layouts, filter selection, interval and injection times, as well as measurement frequency, heating, and shaking parameters. During measurement, it is possible to observe the data with the Real Time well display feature, which allows viewing of raw values, ratios, absorbance spectra, kinetic curves, or even color coded pictorials.
The new proprietary developed evaluation software allows displaying and analyzing endpoint, kinetic and spectral data using comprehensive data reduction algorithms and various curve fitting options. Up to 384 complete spectra or kinetic curves can be viewed individually or overlaid. For further data processing results can be easily exported to Microsoft Excel™.
Windows 7/ XP/ 2000 and Vista are a trademark of Microsoft Corporation.< br />DELFIA® is a registered trademark of PerkinElmer, Inc.
|Detection modes: || UV/Vis absorbance spectra |
Luminescence (flash and glow)
|Measurement modes: || Endpoint and kinetic measurements |
Sequential dual excitation measurements
Simultaneous dual emission measurements
|Light source: || Xenon flash lamp |
|Detector: || Two matched side window photomultiplier tubes |
|Filters: || Excitation and emission filter wheels for 8 filters each |
|Wavelength range: || 240 - 1000nm |
|Reagent Injection: || Up to two built-in reagent injectors |
Injection at measurement position (6 to 384-well)
Individual injection volumes for each well (3 to 350 µL)
Variable injection speed (100 to 420 µL/s)
Up to four injection events per well
|Temperature Control: || + 5°C above ambient to 45°C (60°C optional) |
Temperature stability: 0.2°C
Temperature gradient: < 0.5°C
|Shaking: || Linear, orbital and double orbital |
|Microplate Formats: || 6- to 1536-well plates |
|Sensitivity: || FI: < 0.2 fmol/well sodium fluorescein |
FP: < 5 mP SD at 1 nM sodium fluorescein
TRF: < 30 amol/well europium
LUM: < 30 amol/well ATP
|Reading Speed (96-well): || 9s |
|Reading Speed (384-well): || 16s |
|Special: || Simultaneous Dual Emission Detection of two signals for FP, BRET, and FRET assays |