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Fluorescence Imaging Workflow

  1. Insert objective; select matching Objective in Micro‑Manager (keep metadata accurate).
  2. Set filter layer to Multiband (DAPI/FITC/TRITC).
  3. Place the slide and cover with the stage cover to reduce ambient light.
  4. Select Kikker camera; Auto Shutter on; set mirror slider right (to Kikker).
  5. In Channels, set OFF initially; click Live.
  6. On IRIS‑400, select R/G/B/UV wavelength; adjust intensity; press the button to turn ON.
  7. Increase exposure until visible (100–800 ms typical). Start with longer wavelengths to reduce photodamage.
  8. Focus; refine exposure to maximize contrast without clipping. In 12‑bit mono, avoid maxed pixels at 4095.
  9. Save via Save or Stop Live → Snap → Save.