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Detection windows available:

  • 430-520 nm  (blue)
  • 500-550 nm  (green)
  • 570-650 nm  (red)
  • 665-760 nm  (far-red)
Tipp

What about my specific excitation line?
For fluorophores with an excitation peak at for example 510, use the closest available light source: 475 nm

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Tipp

Use overview image to pick interesting regions
Using "Test Images", and marking regions, right-click and "use as background for well" for a visual aid for selecting further regions.

Run the experiment:

  • Evaluate that the correct assay is loaded.
  • Name the plate and add keywords.
  • → Start! And observe live-preview images during acquisition (when enabled)
  • Select the whole plate, right-click, organise images in "realistic" (corresponding to the plate) or "packed" (best for display) layoutsinfo:
Tipp

Flat-field correction
The software can do flat-field correction based on acquired images, provided that foreground/background and areas without cells are present.
This also works in wells filled with tissue, or more-than-confluent cell cultures.

  • When done: open the lid (settings - Operetta CLS - open) and remove plate/slide holder.

Image processing and analysis:


Image Added





Loose info pieces:


Operetta CLS: zZ-steps always > 100 nm


Fluorescent staining strategy,:

To facilitate auto-detection in subsequent image analysis steps:

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c) stain nuclei separately


Computer attached to the microscope:

DO NOT SWITCH OFF!

The PC is running a live image data and image analysis server.

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