Protocol: Converting Time Lapse Images into Movies Using Photoshop CS4

Load Raw Images

• Navigate to folder containing time lapse images using Adobe Bridge
• Highlight all images in series
• Tools…Photoshop…Load Files into Photoshop Layers…

Adjust LUTs and Sharpen Images

• Image…Auto Tone
  • NOTE: If adjustment looks incorrect, use Image…Adjustments…Levels, for finer control
  • Filter…Sharpen…Smart Sharpen…
  • Increase the Amount to approx 200%. Increase the radius by increments of 1.0 until there is a good tradeoff between the object of interest and the background noise. Ensure “more accurate” is selected
  • Hide the layer
  • Repeat adjustment steps for remaining layers
    • NOTE: This can be done more efficiently by using actions (see Below)

Align Image Stack

• Make the last two image layers visible
• Select the higher layer and in the LAYERS panel, set the opacity to 50%
• Select the move tool and shift the upper layer so that it aligns with the layer below
  • NOTE: If the image seems to skip to incorrect positions automatically and you want more control go to View … Snap, and ensure it is deselected
  • To ensure alignment is accurate, toggle opacity between 50% and 100%. Once satisfied leave the upper image opaque.
  • Select the next layer above and repeat the alignment process

Crop Image Stack

• Once all images are aligned, Select …All Layers, and zoom out so that the entire image is visible
• Using the crop tool select only the region of the stack that is common to all images
  • NOTE: To get a rough idea of where this region is, scroll through the LAYERS panel and plan to remove all of the checkered grey/white regions
  • Hit enter to apply crop

Export Layers to Images

• File…Scripts…Export Layers to Files…
• Set a destination folder and an image format (TIFF/JPEG)

Convert Images to Film

• Using ImageJ, File…Import…Image Sequence, navigate to your exported images
• Ensure the images are well aligned by moving the horizontal slider back and forth
• File…Save As…Avi (settings = uncompressed and increase/decrease frame rate as required)