The DeltaVision OMX V4 from Applied Precision (GE Healthcare) is a fully automated inverted microscope - objectives are below the stage. The OMX is a multi-mode imaging platform that enables imaging of fixed or live specimens at better spatial resolution than conventional microscopy (wide-field and confocal microscopy).
![Super-resolution Deltavision OMX V4](/~/media/Images/core-facilities/CIC/2020-equipment/Super-resolution1.ashx?la=en)
Due to the wave nature of light, any point source of light (even from a single GFP molecule with a size of 3 nm) is observed by the microscope as a small diffracted blur. As a result, the spatial resolution (i.e. ability to distinguish 2 points) is limited to ~250 nm in the x-y plane and ~600 nm in the z-direction. Compared to conventional microscopy, the OMX super-resolution microscope improves spatial resolution by a factor of 2 to 5.
The OMX is equipped with a structured illumination microscopy (SIM) module, which enables 3D super-resolution imaging of live and fixed samples at ~120 nm lateral and ~300 nm axial resolution using standard fluorophores.
The OMX also contains a DeltaVision localization microscopy (DLM) module, which achieves imaging of fixed samples at a 20-50 nm lateral and 100-150 nm axial resolution using photoactivatable and photoswitchable fluorophores.
The OMX imaging platform also offers TIRF (Total Internal Reflection Fluorescence) microscopy, which illuminates fluorophores within ~100 nm depth from the coverslip-sample interface without interference from out-of-focus light.
For live cell imaging, a “Ultimate Focus” module maintains samples at the focal plane over time, and a temperature-controlled chamber and objective warmer maintain optimal environmental conditions.
A better spacial resolution, but what for?
Super-resolution imaging techniques have become commercially available in the past 5 years and their use will continue to grow exponentially. Super-resolution is revolutionizing many different biological and biomedical research fields. The reason why is simple: we can now visualize and study molecular assemblies and structures that were not accessible to conventional light microscopes.
Examples of applications include the study of micro-organisms (e.g. bacteria, viruses), organelles (nucleus, mitochondria, vesicles), protrusions (filopodia, spines, cilia), plasma membrane organization (lipid rafts), and structural organization of large protein complexes (cytoskeleton, synaptic protein, nuclear pore complex).
![Microvilli WF vs SIM](/~/media/Images/core-facilities/CIC/equip-subpage-photos/deltaomx.ashx?la=en)
Light Sources
Halogen lamp for bright-field microscopy (i.e. white light illumination to acquire a contrast image)
Solid state illumination module for wide-field fluorescence microscopy capable of simultaneous operation of 4 user-selectable wavelengths from 6 possible excitation wavelengths (405, 445, 488, 514, 568, and 642nm)
Laser lines for TIRF, 3D SIM, and Localization microscopy:
- 405 nm for DAPI staining
- 445 nm for CFP / Alexa 430
- 488 nm for GFP / FITC / Alexa 488 / Cy2
- 514 nm for YFP
- 568 nm for tdTomato / mCherry / TRITC / Alexa 568 / Cy3
- 642 nm for Alexa 647 / Cy5
Objectives
(Magnification / Numerical aperture)
- 60x/1.42 (Olympus)
- 60x/1.49 (Olympus) – for TIRF
Detector
3 pco.edge sCMOS cameras (custom version for DeltaVision OMX V4)
Acquisition software
OMX Master (GE)
Image reconstruction and analysis software
softWoRx 6.1.1 (GE)