ImageXpress Micro Confocal

Molecular Devices ImageXpress Micro Confocal

  • An inverted fluorescent microscope with a transmitted light option for label-free imaging. 
  • Features a 60 uM pinhole single spinning disc for confocal imaging. 
  • Widefield imaging option for high-throughput screening. 
  • High-quantum efficiency sCMOS detection camera with a large field of view

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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.

ImageXpress C Objective Options

There are 4 objective positions within the IXM-C, the standard instrument configuration has the 4x, 10x, 20x, and 40x objectives in the instrument (bold in the table below). If you are interested in using one of the objectives not installed in the instrument, please contact the screening facility with your request before your scheduled usage time. 

  • The 2x objective captures the whole well of a 96-well microplate.
  • The 4x objective captures the full well of a 384-well microplate, or four 1536-microplate wells at a time. 
  • The 10x objective captures a whole well in a 1536-well microplate.  
MagnificationObjective TypeNA (uM)WD (mm)
2XPlan Apo λ0.108.5
4XS Fluor0.2015.5
10XPlan Fluor0.3016
10XPlan Apo0.454
20XS Plan Fluor *0.458.2-6.9
20XPlan Apo0.751
40XPlan Apo λ *0.950.25-0.17

* These objectives have a correction collar

ImageXpress C Filters

For excitation wavelengths, instead of using traditional excitation filters or lasers, the IXM-C uses a power-optimized solid-state light source. In addition there is a 5 position dichroic wheel, and an 8 position emission filter wheel. 

Fluorophores Excitation (nm) Emission (nm) Dichroic (nm)
DAPI, Hoechst 380-420  417-477 409
FITC, GFP 440-500 516-556 506
CY3, TRITC 490-550 573-613 562
Texas Red, mCherry 530-610 604-644 593
CY5 610-660 672-712  660