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August 20, 2002
Olympus Launches MF20 System
for Analysis of Biomolecular Interactions
Single molecular fluorescence spectroscopy enables analysis
of protein interactions in solution in just a few seconds
MF20 molecular interaction analysis system
MF20 molecular interaction analysis system
Olympus Optical Co., Ltd. (President: Tsuyoshi Kikukawa) has completed the joint development of the MF20 molecular interaction analytical system with Evotec Technologies GmbH (Hamburg, Germany; CEO: Dr. Carsten Claussen). The MF20 system can analyze molecular interactions, thus providing valuable clues about protein function. This promises to help in the detailed elucidation of biological phenomena, and to play an important role in genome-based drug discovery. The company plans to start taking orders for the system in Japan from the end of August 2002 and will make the product available in overseas markets in due course. The MF20 system employs single molecular fluorescence spectroscopy (Note 1), a technology that not only permits high-speed measurements relative to existing techniques, but also facilitates inexpensive, high-precision analysis. The principal target market for the system comprises pharmaceutical firms and other drug research companies, who could use the MF20 in receptor (Note 2) studies to search for ligands (Note 3) with optimal binding characteristics. Other potential users include universities and research institutions.
Main Features
  1. High-speed analysis of protein or DNA interactions with high sensitivity and low noise
  2. High-throughput analysis possible
  3. Low running costs
  4. Various analytical modes simultaneously available on single system
  5. No pre-analysis sample preparation or washing required
  6. No use of solid-phase bioassay, thus eliminating reliance on sensor chips
Product Background
Following the elucidation of the broad outline of the human genome, the focus of genomic research has switched to the analysis of protein function (proteomics). Many researchers in the field are now actively searching for ligands and other substances that interact with proteins. There has therefore been a strong demand within the market for equipment that can facilitate the analysis of biomolecular function under conditions close to those that exist in the human body.

The MF20 employs a technique known as single molecular fluorescence spectroscopy that does not require the sample to be in a solid phase. Instead, interactions between biological molecules can be studied directly in buffer solution. High-speed analysis yields data on functional biomolecular interactions and binding within a short period of time ranging from a few seconds to less than a minute. A combination of confocal laser optical system (Note 4) with a high-sensitivity fluorescence detection device permits measurements in an extremely small volume of around 1 femtoliter (1 fL = 1 x 10-15 L), thereby capturing any interactions at the level of single molecules. This set-up operates at low noise and detects fluorescent signals with extremely high sensitivity. In addition, since measurements are performed in solution, damage to the sample is minimal and artefacts from unspecific surface interaction are avoided. This allows the same sample to be rapidly put through other tests subsequently.

The MF20 system comes equipped with three standard analytical modes that provide different methods for studying samples-FCS (Note 5), FIMDA (Note 6), and FxCS (Note 7). It can be used in various applications, from high-throughput analysis to complex studies of molecular interactions. The system promises to make a significant contribution to the detailed elucidation of biological phenomena, thereby playing an important role in genome-based drug discovery.

Details of Main Features
  High-speed analysis of protein or DNA interactions with high sensitivity and low noise
    The system enables reduced noise and highly sensitive measurement of the fluorescent signal (over 0.1 nmol/L) by means of the combination of confocal laser optical system with a high-sensitivity fluorescence detection device (Avalanche Photodiode). This permits measurements in an extremely small volume of solution of around 1 femtoliter (1 fL = 1 x 10-15 L), thus capturing any interactions at the level of single molecules.
  High-throughput analysis possible
    Analysis of a single sample is complete within a short period of time ranging from a few seconds to less than a minute. The MF20 system can thus be used in applications ranging from high-throughput analysis to complex studies of molecular interaction.
  Low running costs
    The system design minimizes the need for reagents and other consumables (such as glass-bottomed microplates and distilled water), thereby keeping running costs low.
  Various analytical modes simultaneously available on single system
    The MF20 is equipped with FCS, FIMDA, and FxCS analytical modes as standard features. These permit the analysis of various molecular parameters, including size, concentration, fluorescence brightness, and the respective amounts of different molecules present. The system can also be applied to various types of analysis that were previously difficult to undertake, such as that of interactions between molecules of similar weight or simultaneous measurements at two different wavelengths.
  No pre-analysis sample preparation or washing required
    The MF20 system can simultaneously measure and distinguish between the fluorescent signals of both, bound and free molecules in a solution composed of both types. The time-consuming yet vital sample preparation and washing steps required with previous analytical methods are thus rendered unnecessary.
  No use of solid-phase bioassay, thus eliminating reliance on sensor chips
    Operation involves the injection of the reaction sample solution into a glass-bottomed microplate that is then loaded into the device. After these simple steps, analysis is completely automated. There is no need to undertake any difficult preparatory steps to transfer the sample onto a solid phase carrier such as a sensor chip or microplate well bottom, a process that is required with other analytical methods such as ELISA (Note 8) or Surface Plasmon Resonance (Note 9). In addition, tedious coating or regeneration procedures, which are required for most solid phase assay techniques are not necessary at all with the MF 20 system.
    (NOTES)
    1. Single molecule fluorescence spectroscopy
    Through the use of confocal laser optical system, this technology enables the measurement at the level of single molecules of fluorescent-labeled molecules. For example, a fluorescently labeled ligand will interact with specific receptors. The ligand-receptor complex will take more time to diffuse across the focused laser beam and thus generate a different signal from a free ligans.
Another example of molecular interaction is the detection of DNA molecules. They can be detected by the use of fluorescently labeled "primers" integrated to the template strands through an amplification method such as PCR (Polymerase Chain Reaction: the primers are DNA chains with nucleic acid sequences that are complementary to the template DNA which has to be measured). Such measurements can capture the behavior of biological molecules in extremely small volumes, measured in femtoliters (1 fL = 1 x 10-15 L), by analyzing the size of the amplified DNA molecules or of proteins, as well as the number of molecules within the measurement field. In this way, molecular interactions can be studied in solution close to the level at which molecules such as proteins and DNA work. The system was developed jointly with Evotec Technologies GmbH (Hamburg, Germany; CEO: Dr. Carsten Claussen).
    2. Receptors
    Proteins inside a cell that are used to translate specific physical or chemical stimuli into a certain response by the cell.
    3. Ligands
    Proteins, peptides, or other substances such as drugs that can transmit information by binding to other molecules, e.g., receptors.
    4. Confocal laser optical system
    A system of optics that uses a pinhole at the point of focus in order to exclude all light except that from the focal position. This arrangement generates much better S/N (signal-to-noise) ratios than those produced by conventional methods for reading fluorescent signals.
    5. FCS (Fluorescence Correlation Spectroscopy)
    A method for studying multiple molecular parameters (such as translational diffusion time (Note 10), which indicates molecular size, or molecular concentrations) for various types of molecules using analysis of fluorescent signals.
    6. FIMDA (Fluorescence brightness Multiple Distribution Analysis)
    A calculation method that measures, in addition to translational diffusion time and molecular concentration, the fluorescence brightness per molecule.
    7. FxCS (Fluorescence Cross-Correlation Spectroscopy)
    A calculation method that analyzes fluorescent signals from molecules labeled with two different dyes to determine whether molecules are moving individually or bound together. This permits highly sensitive analysis of molecular interactions, which is particularly useful with types of analysis that were previously difficult to undertake, such as that of interactions between molecules of similar weight.
    8. ELISA (Enzyme-linked immuno-sorbent assay)
    A type of solid-phase enzyme immunoassay, ELISA involves labeling antigens or antibodies with enzymes and then using an enzyme reaction to detect the presence of either antibodies or antigens.
    9. Surface Plasmon Resonance (SPR)
    A method that analyzes binding and release between two molecules by optically detecting as an SPR signal any minute changes in mass in the sample, which is studied on the surface of a solid-phase sensor chip.
    10. Translational diffusion time
    A physical coefficient for a molecule that increases in line with its molecular size.
Overview of Evotec Technologies GmbH
Address : Schnackenburgallee 114, 22525 Hamburg, Germany
President & CEO : Dr. Carsten Claussen
Year of foundation : 2002, before division of EvotecOAI AG, founded 1993
Nature of business : Evotec Technologies offers innovative solutions for complex life science applications. The company provides seamless integration of hardware, software and bioware modules, combining cutting-edge technologies for measurement, miniaturization and automation. It employs around 60 people at three locations in Germany (Hamburg, Dusseldorf and Berlin).
URL : http://www.evotec-technologies.com/
Main Specifications
Analysis Method
Single molecular fluorescence spectroscopy (FCS, FIMDA, FxCS)
Measurement Parameters
Concentration, translational diffusion time, molecular fluorescence brightness, correlation coefficients
Laser Wavelengths
488nm, 543 nm, 633 nm (simultaneous measurement possible at two wavelengths)
External Appearance
Bench-top device measuring W710mm x H445mm x D600mm(argon laser affixed to exterior)
Weight
Approx. 120kg
Automated Adjustment
Focal position, confocal optical system, correction of objective lens
Power
Input voltage = single-phase AC 100V-240V;
power consumption rated at 600VA
(argon laser requires separate power source rated at 2000VA)
Software
Proprietary software running on Windows 2000 O/S
Accessories
PC, monitor
Consumables
Glass-bottomed microplate (96,384 wells/plate), distilled water
*Olympus Optical Co., Ltd. was changed to OLYMPUS CORPORATION as of October 1, 2003.
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