Acumen

Laser Scanning Cytometer

The TTPLabTech Acumen eX3/HCI (High Content Imager) was purchased with funding from Harvard Medical School’s Tools and Technologies Committee.

  • Bottom-reading fluorescent imager
  • 3 excitation lasers (405, 488 & 561 nm)
  • Detect up to 4 different emission wavelengths simultaneously.
  • Read and analyze a 384-well microplate in as little as 7 minutes (1 laser ex., up to 4 channel em, 0.5um/pixel resolution).
  • Capable of imaging and analyzing 3D cultures and whole organisms due to a large depth of field.
  • Two virtual access analysis workstations with Cellista software.
  • Additional information can be found on the TTP LabTech website.

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Designing an Image-Based Screen

Assay plates:

Black-walled, clear-bottom (plastic or glass) microplates are used for image-based screens. A common 384-well plate type is Corning #3764 (previously #3712).

Seeding density:

Test a variety of cell densities under the conditions you plan to use for the screen. Image analysis is often improved when the cells are sub-confluents. Depending upon cell size and length of assay, a starting range may be 500 to 2,000 cells per well.

Imaging live cells:

This is possible when using the Acumen. Remember that the microscope stage is not heated and your cells will be at room temperature during imaging.

Treating plates after imaging:

After processing cells for imaging, fill your wells with PBS to prevent the cells from drying out and to reduce background noise. Some investigators include an anti-fungal agent (eg sodium azide) for long term storage at 4oC.

Selecting fluorophores:

Please make sure the fluorophores you select do not conflict with the filter sets on the screening scope. See details for each microscope. For high throughput screens, antibodies are frequently used at higher concentrations to decrease acquisition time.

Filters

Wavelength (nm) Dichroic (nm)
420 490
500-530 540
575-640 640
655-705 Mirror