BioKine32 is BioLogic's powerful and user-friendly software platform for instrument control, data acquisition and analysis of recorded spectral and kinetics traces. It is running under Microsoft Win XP on any recent personal computer.

Instrument control

Control of Stopped-Flow mixers

BioKine32 is used to control all actual stopped-flow and quench-flow mixer models: SFM-20, SFM-300, SFM-400, QFM-400. Also older models can be controlled, such as SFM-3, SFM-4, and QFM-5.

Configuration of stopped-flow mixers and accessories

Filling and purging of syringes is performed using the syringe command window. The filling speed can be controlled manually in six steps in order to adapt operation to washing and purging procedures (high speed) or filling of very small volumes (low speed). Due to the stepping-motor technique, the exact actual filling volume of all syringes is displayed continuously at any time during syringe movement. In the shot mode, the number of left-over shots is displayed and provides the user with useful information to plan his experimental schedule and autorun sequences.

Two modes (advanced and classical) allow an easy and straightforward programming of mixing sequences from one simple mixing step up to more complex mixing events such as intermediate ageing (sequential mixing), concentration-dependent series or quench-flow application. Also automated titration sequences with variable user-defined volume or concentration steps can be designed in combination with the titrator accessory.

Advanced programming of a mixing sequence for protein folding studies (SFM-300 with micro-cuvette, 1:10 dilution of a denatured protein from 6 M to 0.6 M Guanidine-HCl, estimated dead time: 0.3 ms).

Classical programming of a mixing sequence for a double-jump protein folding experiment (SFM-300 with micro-cuvette); first jump: partial unfolding of protein from 0 M to 6 M Guanidine-HCl during 100 ms (72.2 ms + 27.8 ms), second jump: protein refolding from 6 M to 1 M Guanidine (estimated dead time: 0.4 ms).

Start window for programming of automated titration sequences.

• Control of detection devices

Besides the control of BioLogic's stopped-flow and quench-flow mixer modules, BioKine32 allows also to control data acquisition from a large variety of detection devices, such as: MOS-200, MOS-250, Diode array, and JASCO J-810 spectropolarimeters, as well as external devices such as FP-6500.

Installation of detection device

J-810 control and setup window

Data Acquisition

The Bio-Kine32 software can control the stopped-flow mixers and detection devices described above and acquire data for transient kinetics, spectral scans, and act as a virtual chart recorder. The use of similar controls for all modes makes Bio-Kine32 easy to use and eliminates the hassles of learning multiple programs for different types of experiments.

Transient Recorder (kinetics)

The Transient Recorder is similar to a digital oscilloscope. Up four data channels of 8000 points each can be acquired and displayed simultaneously. Sampling rates from 10 µs/point (100 kHz) to 100 s/point (0.1 Hz) can be used. Both linear and logarithmic sampling is available through the use of up to three times bases. This allows to optimise data acquisition (integration times) for multi-exponential kinetics traces. Data can be acquired in two different spectroscopic modes simultaneously (Volt, Transmittance, Absorbance, CD or Anisotropy).

Transient recorder (kinetics): example of setup for data acquisition with a MOS-200 spectrometer

Scanning Spectrophotometer

The Scanning Spectrometer is used to control one or two monochromators to perform spectral scans. Bio-Kine32 is capable of making spectral acquisitions in the same spectroscopic modes that are available for transient kinetics. In conjunction with the appropriate spectrometer system, Bio-Kine32 can act as a full feature absorbance, fluorescence or CD spectrometer.

Scanning spectrometer: example of setup for data acquisition with a MOS-250 spectrofluorimeter

Global mode

Some detection devices (MOS-200/M and MOS-250) offer the possibility to record 3D-kinetics in a global mode, i.e. to make one shot (or an average of several shots) per wavelength and subsequently increment the actual wavelength in defined steps. This allow to obtain a three-dimensional kinetics with the signal as a function of time and of wavelength without the use of a diode array detector.

Scan kinetic (global mode) : example of setup for data acquisition with a MOS-200/M spectrometer

Scan kinetic (global mode) : example of shot control and data acquisition with a MOS-200/M spectrometer

Chart Recorder

The Chart Recorder is similar to the Transient Recorder, but specialized for slower acquisitions over long periods of time. Up to four data channels and two spectroscopic modes can be recorded simultaneously. Data is displayed in real time, and scrolls along acquisition window. Files of several million points can be recorded with event markers being added at anytime.

Data analysis

BioKine32 provides a waste number of tools for data analysis and data treatment:

• Multi-exponential fitting of kinetics using a simplex algorithm
• Second order fitting of kinetics
• fitting with user-defined functions
• average, integral and rms noise of a defined portion of the data trace
• Baseline calculations and differences
• data smoothing
• display and analysis of multi-wave length kinetics (global baseline, multi-extract)
• mathematical operations with one trace (factor and offset), two traces (difference, sum ratio, average) or up to 10 traces (average)

Analysis Tools in BioKine32

Math options in BioKine32

Example of fitting of a fast kinetics trace with a single-exponential function (data yellow; fit green). Please note that start of data acquisition was 20 ms before the stop, thus, the kinetics traces starts at 0.02 s (marker “0”). In the upper part, residuals between data trace an fit is displayed in green.