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Dual-Channel Acquisition

Two fluorescence channels for nuclear reference plus reporter

Definition
Simultaneous or sequential capture of two fluorescence channels, typically DAPI for nuclei and GFP for a reporter. The two images are co-registered, so each nucleus can be classified as marker-positive or negative.
Co-Registered
Both channels captured at identical positions
Filter Switching
Automatic excitation and emission filter changes

The Two-Channel Configuration

The most common PanoBrain configuration captures two channels per section:

  • Channel 1 (DAPI): Labels all nuclei, providing the reference for cell detection and total cell counts
  • Channel 2 (GFP or another reporter): Labels the population of interest (transduced neurons, transgenic cells, activity-dependent expression)

Because both channels are captured at the same tile positions, they are co-registered by construction. Each DAPI nucleus can be checked directly for GFP signal without aligning the images.

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