A unique Software Package for Fluorescence Spectroscopy
Fluorescence spectroscopy is one of the most commonly used spectroscopic techniques in modern biochemistry and molecular biophysics research. No other technique can offer as much in terms of sensitivity and selectivity with such moderate operational costs.
Fluorescence measurements do not yield such detailed structural information as infrared spectroscopy, for example, nonetheless the technique has become extremely popular due to its high sensitivity and selectivity, which can be used to dynamically follow structural and dynamic properties of physiological importance in intact living systems while using probe concentrations low enough to avoid perturbing the system. Fluorescence is also widely used in solid state measurements, in industry and in pharmaceutical Quality Control to great effect.
The range of applications and measurement types in fluorescence spectroscopy is huge. Measurements can be carried out at the simplest level, collecting simple fixed wavelength intensity data for quantitation, through kinetic measurement of biomolecular interactions to 3-Dimensional spectral studies to help resolve complex samples.
An important factor in the use of fluorescence spectroscopy is the wide range of sampling accessories which are available for handling diverse sample types and for the automation of multiple sample measurement.
Equally important is the functionality of the software used to control fluorescence measurements. On the one hand this must be easy to use for simple, routine measurements. On the other hand, the software must be able to handle more complex and demanding automated experiments, both in terms of sample handling and data calculations and calibration.
Ease of use
The BioLight Studio Software package was designed for intuitive operation from the very start:
At the most direct level, this offers the researcher an easy-to-use, walk up system. Open a method, click on ‘Start’, and you’re up and running. Measure samples till you are finished, then click on stop. All data from the current sample batch is saved automatically in a single multifile: No more searching the hard disk to find which data belongs to an experiment, because all the data from the current experiment is stored conveniently in one file. Additionally all data can easily be copied and pasted into Word or Excel.
At the other extreme, BioLight Studio allows the user to add sophisticated programmable calculations, automated concentration calibrations, and detailed but easily user-designed reports. And all steps in between. And all of this can be automatically applied to all samples within a batch for high throughput, routine users. In the basic version of BioLight Studio all records are stored in a file system. Thus it is possible to use the Windows file explorer to easily identify and archive results and methods.
Integrated Sample Handling
All sample information are defined in a customizable table structure. Specific sample information like a dilution factor can easily be added to the table. This information can be used in calculations and be inserted into reports.
When a series of samples is to be measured, the Samples page is used to define the number and type (Blank, Reference, Unknown). The Samples page is also used to define sample positions for automated accessories such as the Plate Reader or Autosampler. Figure 2 shows the Samples page for a set of 6 standards and 18 unknown samples:
Sampling accessories (for example, the Plate Reader Autosampler or Peltier temperature controller) are fully integrated into all data acquisition types in the system. This means that the user can collect spectra or even 3D EEM spectra from selected wells in a microtitre plate for instance, or to automatically sample and quantitate samples using the Autosampler.
Setting Instrument Parameters - it can't be easier
Instruments parameters are visualized showing the light-path diagram, together with the relevant instrument components.
So it is very straight-forward to define the scan wavelengths, scan speeds, the light sources, the slit settings, the cell holders and the measurment mode and other instrument settings.
These parameters are accessible in the "Acquisition" tab of the Biolight Studio Software Package.
The basic version BioLight Studio offers 3 calculation modules that allow to perform an automated data processing during the measurement: The standard calculator, the concentration calculator and the 3D viewer.
The screen shot to the right shows a series of scans, where the peak areas and the peak heights were calculated automatically. In addition the readings at certain wavelengths had been extracted. To obtain spectral purity information the ratio at certain wavelengths had been calculated. Finally, even, a decision was introduced. In the case that the calculated ratio is below a certain given threshold an information will be output, to tell the researcher that the substance is not anymore spectrally pure.
The screen shot to the left displays the concentration calculator. In this example a calibration curve based on 4 references was calculated and displayed. The calculated concentration is shown for each of the six measured samples, together with the sample ID, the measured signal, and dilution factor.
All calculators allow the user to easily copy the results, either as tabular data or as vector-graphic, to the clipboard.
From there they can be pasted directly into word processing or table calculation programs for further evaluation and reporting. Of course all results can be inserted into the BioLight Studio report module without the need for an external program.
In addition to the automatable calculation modules, a stand-alone calculator is delivered free of charge. Your colleagues can analyse and re-process raw data and results off-line, free of charge.
The BioLight Studio report system is very intuitive and easy to use, offering a comprehensive array of information which can be quickly added to the report template.
Even for the casual user, a report can be produced which precisely describes all aspects of the experim-ent. Quickly.
The example to the left shows a report for the measurement of Nitrate concentration in contaminated water. Six standards and three unknown samples were measured. The report includes the relevant measurement parameters and a calibration curve which displays the position of standards and samples within the curve. The raw data, together with the calculated concentrations, are displayed together with sample name and sample information.
BioLight Studio stores the report template together with the raw data, rather than the final report, thus allowing the user to add or remove results, tables and graphs and re-generate the report whenever required. The final reports can be printed or exported in various formats, including the .pdf format.
21 CFR Part 11 Support
In the basic version BL Studio identifies the current user from the Windows Login. It is possible to assign different method and data directories for each user.
All data are stored in binary records, secured with a checksum. To ensure the reproducibility of results all raw data are stored together with the acquisition methods, all calculation parameters and the report template.
On opening a Biolight Studio data set, the method used to collect the data is automatically re-loaded in its entirety. New data can be measured using exactly the same conditions as before.
All methods, results, configuration data and protocol records support electronic signatures. In the basic version electronic signatures are added automatically whenever new results or methods are stored.
To allow for the traceability of results BL Studio offers a comprehensive audit trail. The audit trail is organized as one record per day, thus allowing to easily archive data!
The Verification module is provided with the BL Studio, to allow the user to regularly check instrument performance. The Validation module automatically carries out a series of tests to determine the instrument’s performance in terms of wavelength accuracy, sensitivity and drift.
Data is displayed in real-time, and the results are automatically saved for later archiving. A report can be printed, or copied to the clipboard for further processing with a word processing program.
BioLight are constantly developing new, plug-in applications. These can be easily added to the system individually: The system structure remains unchanged. This means minimal re-validation, no down-time and instant expansion of the Biolight Studio.