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Camera

The Hamamatsu ORCA-Fusion BT in NIS-Elements offers different options for bit-depth and readout speed. The correct choice will be determined by the experiment.

Bit Depth#

Bit-depth option Dynamic range Offset Typical use case
12-bit Sensitive Low 100 Live-cell
12-bit Medium 0 Fixed
16-bit High 100 Fixed

Scan Mode#

Scan mode Readout noise Readout speed Typical use case
Ultra-quiet Very low Slow Fixed imaging 1
Standard Low-medium Medium Live-cell 2
Fast High Fast Very fast imaging3

ROI#

  • The CSU-X1 limits the field of view to a max of 7x10mm. With a 100x objective and no other sources of magnification, the field of view roughly corresponds to 70um by 100um.
  • Saved camera ROIs are saved in the Documents folder and can be loaded by clicking on the "ROI Size" button and pressing "Load ROI..."

Laser illumination is not even across the field of view

  • If you care about intensity measurements or will be using intensities for thresholding, include a "flat field correction" processing step.
  • If acquiring tiled images, consider using the multipoints tab for grid selection and stitching after correcting each tile with BaSiC or dye solutions4.

  1. Permissible for live-cell imaging if acquiring single images with a delay between timepoints longer than the readout time ↩

  2. Or fixed imaging with bright signal and wanting to save time on tiled z-stacks ↩

  3. When 10s of milliseconds matters ↩

  4. Contact Will for the dyes ↩