#  Acumen 

 



## Laser Scanning Cytometer

 ![Acumen Laser Scanning Cytometer](/sites/g/files/omnuum5406/files/iccb/files/acumen.png)

 

- Bottom-reading fluorescent imager
- 3 excitation lasers (405, 488 & 561 nm)
- Detect up to 4 different emission wavelengths simultaneously.
- Large depth of field, enabling imaging and analysis of 3D cultures and whole organisms.
- Integrated with a Thermo Orbitor, enabling walk away capability for 60+ assay plates.

The [STP Labtech](https://www.sptlabtech.com/) Acumen eX3/HCI (High Content Imager) was purchased with funding from Harvard Medical School’s Tools and Technologies Committee.



 

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###    Designing an Image-Based Screen  expand\_more  

 

####  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](/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.

####  Plate Seals:

 Clear and foil microplate seals can significantly increase background fluorescence to the point where images may be unusable. Black opaque plate seals are ideal for fluorescence imaging. Microplates must be sealed prior to imaging in the microscope to protect the equipment.



 

 

 



 

 

 

 

##  Filters 

SortWavelength (nm)Dichroic (nm)420

490

500-530

540

575-640

640

655-705

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