- always choose file format first;

- accepted file formats:

(a) Bruker *.par and *.spc files (OS-9 format);

(b) generic one column ASCII;

(c) generic two column ASCII (x,y) - make sure that the delimiter between the columns is correctly set in "ASCII File Read 2 Columns"  routine (in PF.llb library);

(d) PF ASCII format:   one column ASCII, first row is number of points;

(e) Global Lab "prf" format of gel scans;

(f)  Lambda 50 fluorometer spectral files;

- make sure that the cursor order on the graphs follows the order on the legend below the graphs;

- most programs allow for following baseline corrections:

(a) single horisontal line defined by first cursor;

(b) two points sloping line defined by first and fourth cursor;

(c) range averaging: as in (b) but the extreme points are defined by the spectral averages in between the 1 & 2 cursor on the left and 3 & 4 cursor on the right;

(d) no baseline subtraction;

- all spectra are automatically stored in the HPGL format for import to Powerpoint ot Word, name "*.plt" where "*" is input file name root;

- most programs allow for storage of the manipulated spectra in PF_ASCII format;

- MATLAB has to have the LABVIEW-EPR directory added to its path !

- fits can be to following functions:

(a) Gaussians;

(b) Lorentzians;

(c) cosine square;

(d) straight line;

(e) cubic;

(f) quadratic;

- each function is defined by upto three corresponding parameters e.g. each Gaussian has amplitude, position and width;

- maximum and minimum of the range parameter will be searched for is defined by cursosrs or in boxes;

- initial guess parameter is defined by the middle slide;

- function can be:

(a) single simulation (use for initial guess estimates till ref fit in the panel is more or less the shape of experimental data);

(b) Simplex algorithm:  robust but not efficient;

(c) Levenberg-Marquardt: efficient but guesses must be near the answers;

(d) Newton:  ibido;

- fitting is finished when "green" light light up;

- x-position differences are displayed in small boxes (below the best fir for the first component);

- peaks are stored in Excel fiel (ASCII delimited format) upon request, values are appended to "Peaks.xls" file on the root directory;

-  collects the data on 2 channells from National Instruments A/D board.

- reads the gel image (jpg, tiff formats);

- sets optical density profile along chosen line;

- passes to integration routine;

- reads the parallel and perpendicular intensities (Lambda 50 *.rls files);

- calculates anisotropies with G-factor correction;

- preview of spectra to be averaged;

- current average in the lower panel;

- current slot in a small inset

- two independent spectral regions defined in the upper-right panel;

- each can be fitted with polynomials of 0,1,2,3-rd order:

(a) horisontal line (average) - 0th order;

(b) sloping line - 1st order;

(c) parabola - 2nd order;

(d) cubic fit (good for non symmetric peaks) - 3rd order;

- amplitude dfifference and x-axis difference, amplitude ratio between the peaks is given on the bottom;

- the above values are stored in Excel file (ASCII delimited format) upon request, values are appended to "Peaks.xls" file on the root directory;

- two independent spectral regions defined in the upper-right panel;

- each can be fitted with polynomials of 0,1,2,3-rd order:

(a) horisontal line (average) - 0th order;

(b) sloping line - 1st order;

(c) parabola - 2nd order;

(d) cubic fit (good for non symmetric peaks) - 3rd order;

- peak x-axis and y-axis positions are displayed on the rhs boxes;

- select correct fits (by entering "1" next to peak position) for further processing;

- amplitude dfifference and x-axis difference, amplitude ratio between the peaks is given on the bottom;

- the above values are stored in Excel file (ASCII delimited format) upon request, values are appended to "Peaks.xls" file on the root directory;

- variety of baseline choices

- regions of integration set by cursors

- upper, left panel: original spectrum, set the baseline

- upper, rigtht panel: baseline corrected spectrum, set integration region;

- lower left: first integral - absorption;

- lower right: second integral

- integral is stored in Excel file (ASCII delimited format) upon request, values are appended to "Integrals.xls" file on the root directory;

- spectrum can be shifted;

- new scan range;

- number of points can be divided by 2 (smooth first with Smoothing program);

- smoothing of spectra via i+1 or i+2 spline;

- two experimental spectra are combined according to chosen formula: molar fraction; arbitrary amounts and compared to the third experimental spectrum.

- each spectrum can be shifted horizontally (field) and vertically with respect to other.

- each spectrum can be mulitplied for display;

- the composite spectrum can be stored in a file;

- upper panel displays the four spectra, lower panel shows the difference between the third spectrum and the composite;

- spectra can be independently shifted along x- and y-axis and multiplied;

- stepsize along x-axis can be changed for each spectrum (for comparison of different scan range spectra);

- two spectra can be ovelayed for comparson;

- spectra can be independently shifted along x- and y-axis and multiplied;

- difference spectrum with arbitrary subtraction factor can be created and stored;

- preview of all spectra to be averaged;

- current average in the lower panel;

- preview of current slot in a small inset;

- to average enter 1 in the slot corresponding to a given spectrum;

- a spectrum is displayed with x-axis in Gauss units read from the parameter file;

- horisontal and vertical cursors to aid in a quick measurement of splittings, linewidth or amplitudes;

- two independent spectral regions defined in the upper-right panel;

- each can be fitted with polynomials of 0,1,2,3-rd order:

(a) horisontal line (average) - 0th order;

(b) sloping line - 1st order;

(c) parabola - 2nd order;

(d) cubic fit (good for non symmetric peaks) - 3rd order;

- peak x-axis and y-axis positions are displayed on the rhs boxes;

- select correct fits (by entering "1" next to peak position) for further processing;

- amplitude dfifference and x-axis difference, amplitude ratio between the selected peaks is given on the bottom;

- the above values are stored in Excel file (ASCII delimited format) upon request, values are appended to "Peaks.xls" file on the root directory;

- finds areas under selected spectral regions;

- upper panel: baseline correction definition;

- lower-left: selection of the region to be integrated;

- lower-right: display of the integral;

- integral is stored in Excel file (ASCII delimited format) upon request, values are appended to "Integrals.xls" file on the root directory;

- variety of baseline choices

- regions of integration set by cursors

- upper, left panel: original spectrum, set the baseline

- upper, rigtht panel: baseline corrected spectrum, set integration region;

- lower left: first integral - absorption;

- lower right: second integral

- integral is stored in Excel file (ASCII delimited format) upon request, values are appended to "Integrals.xls" file on the root directory;