CardioPlex
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The analytical objectives of the CardioPlex analysis software are to identify and characterize the variety of dynamic spatial patterns in cardiac tissue. This analysis is made of 3 interfaces.


General Interface

The first user interface is the primary CardioPlex interface (shown at right). It contains two windows for viewing data. The right window, called the Page Display, displays data from all pixels as they are positioned in the array. The Trace Display (left window) provides intensity-versus-time traces from a reduced number of pixels, groupings of pixels, or electrode inputs. Pixels can be selected for the Trace Display by pointing and clicking on the Page Display. A variety of low- and high-pass filters and display adjustments are available for each display. In particular, the two displays can be filtered and processed separately for ease of inspection, before the full spatiotemporal processing of the whole recording. Additional controls are available for averaging over pixels, superimposing, and scaling traces.

Trace Display (Intensity versus Time):
Traces can be displayed as intensity change (I) versus time or as fractional intensity change (I/I) versus time. Segments of the traces may be expanded graphically to focus on critical features.

The Trace Display can display:
1. Traces from individual pixels
2. The spatial averages of selected pixels
3. The FFT of selected traces,
4. The correlation between traces, and
5. Separated or superimposed traces. Superimposed traces can be scaled to equal height for comparing time courses.

Page Display (Spatial Array):
The Page Display can display an array of intensity-versus-time traces for all pixels. The Page Display can also display gray scale or pseudocolor images of the Resting Light Intensity or intensity differences between user selected frames. Multiple groups of pixels may be identified and color-coded on the Page Display; the corresponding individual or average traces appear in the Trace Display. An omit array may be created to exclude selected pixels from the display and analysis. An option is available to replace bad pixels with the average of the four adjacent pixels. The Page Display can display:
1. A map of Traces
2. Differences between user selected frames
3. RMS Noise Levels
4. Light Intensities relative to a Dark Frame (Resting Light Intensity)

Filtering:
The following kinds of temporal high- and/or low- pass filtering are available:
Gaussian,
Butterworth,
Bessel
median,
binomial, and
tau

In addition, spatial filtering options include up to 10 iterations of a 3x3 Median, 3x3 Mean, or 5x5 Gaussian low-pass filter and a variable-width Gaussian high-pass filter.

Other Options:
Both the Page Display and the Trace Display can be exported as TIFF, PostScript, or ASCII files.



Cardio/Movie

The analytical process continues with a Movie of the surface membrane potential, progressing for up to 10240 frames. For immediate review, a grey-scale image of unfiltered data may be scanned by scrolling manually using the Scan feature.

The second analysis interface within CardioPlex is the Cardio/Movie interface. It provides pseudo-color displays of the data (shown at right). Pseudo-color images may be displayed as single frames, multiple frame layouts, and as an animated movie. The color-coded pixel intensities may be displayed directly, as contour plots, or as surfaces in 3D. A variety of color schemes, scaling, and filtering options are available. A trace inset is available to link the current movie frame to a time series for an individual pixel or group of pixels.


Trace Inset:
The Trace Inset supplies an intensity-versus-time plot for an individual or group of pixels selected from the pseudo-color movie frame on display. A red bead on the trace identifies the current frame while the movie is running. A frame may be selected by clicking on the trace inset, as well.

Multiple Frame Layouts:
Primary movie frames may be selected in regular sequences automatically or manually edited to select irregular sequences for simultaneous display in the movie window. Special CardioAnalysis frames may be associated by frame number or frame type in various patterns and accumulated in an editable list for simultaneous display.

Spatial Filtering:
Spatial smoothing and filtering options for the pseudo-color images include up to 10 iterations of a 3x3 Median, 3x3 Mean, or 5x5 Gaussian low-pass filter and a variable-width Gaussian high-pass filter.

Overlay of grey-scale Image:
For anatomical reference, a grey-scale image of the preparation, imported as a TIFF or BMP file, can be overlaid on the pseudo-color images.

Pseudo-Color Scaling:
The pseudo-color scaling of the frames can be manipulated by using different color tables and four different scaling schemes:


The scaling schemes are:
1. Variable (with each detector scaled individually to a standard range);
2. Fixed, using the overall minimum and maximum among all traces;
3. Fixed, using the range of a user-chosen detector for all traces; and
4. Manual.
(When you've tried all these options, it will be clear why it's called pseudo-color - as in "lies, damn lies, and pseudo-color").

Other Options:
The movie can be saved as a movie file and the file can then be read and replayed by CardioPlex. The display on the movie window can be exported (in color or grey-scale) as a TIFF file and a sequences of frames from the movie may be exported as a TIFF-Stack or MPEG file.


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