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Further Information on the OMX 3D-SIM Super-Resolution Microscope
Offering resolution at twice the diffraction limit of traditional microscopes, the DeltaVision|OMX system is the practical solution for three dimensional super resolution imaging that does not depend on special fluorophores or fluorescent proteins. The OMX is also set-up for fast conventional imaging of live cells.
Features of the OMX-SR 3D-SIM Super-Resolution System
- Structured Illumination mode for high-resolution imaging. X,Y resolution to ~ 100nm, Z resolution to ~ 300nm.
- Conventional Illumination mode for fast live cell imaging. Up to 65 frames/second in four colors.
- Objective: 1.4NA Olympus 100X oil.
- Lasers lines for excitation (405, 488, 561, 642) and and emission filters (419-465, 500-550, 609-654, 665-705) for imaging DAPI, green, red and far red fluorophores.
- Four Photometrics Cascade II EMCCD cameras allow simultaneous four color imaging.
- Environmental control for live imaging.
- Controlled by DV-OMX software, image processing by SoftWorx imaging software.
Sample Preparation Guidelines
Everything matters when trying to achieve the best resolution. Care should be taken to optimize sample preparation. The best fixation technique or choice of the best fluorophore for your sample is often an experiment in itself and will be specific to your sample. We are happy to offer advice. Below are a few general guidelines for OMX sample preparation.
- Check the list of excitation lasers and emission filters above and pick fluorophores that will work well with the microscope.
- The specimen should be touching or attached to the coverslip. If possible, fix and stain specimens that are already attached to the coverslip that will be used for the imaging experiment.
- USE ONLY #1.5 COVERSLIPS! This is true for almost all light microscopes but is especially important for the OMX. For best results, the LMIC can supply you with coverslips of the same lot number that we used to calibrate the microscope.
- Mount only one coverslip in the center of the usual (3in x 1in) glass slide. Stage travel is limited to 22mm X 22mm.
- Mounting media matters a lot for this system. We recommend the SlowFade (Molecular Probes) mounting medium because it will create a more uniform optical path and prevent bleaching of the fluorescent probes. The LMIC can supply you with an aliquot of SlowFade. We also have ProLong Gold mountant for long term storage of slides. ProLong Gold hardens over time and as it hardens its refractive index changes. It may take 7-8 days for slides to become fully cured. It is too early to tell how this will effect resolution on the OMX.
- You should seal your coverslips with nail polish. Please make sure it is very dry before you put slides on the microscope. Part of the process of high-resolution imaging is finding the immersion oil that best matches the refractive index of your sample. This may require changing oil several times to find the right one. Cleaning oil off of the coverslip is a lot easier if the coverslip is sealed and fixed in place with nail polish.
As yet there is no manual for the OMX. As guides and helpful documents are created we will post them here. If you have the need or suggestions for other information please contact and let us know at firstname.lastname@example.org.
Publications from our OMX will take some time, be the first to submit an OMX paper and we will list it here.
- Three-dimensional structured illumination microscopy of liver sinusoidal endothelial cell fenestrations.
Cogger VC, McNerney GP, Nyunt T, DeLeve DL, McCourt P, Smedsr?d B, Le Courteur DG.
J. Stuct. Biol. 171(3), p382-388, 2010 (PubMed)
- Fast live simultaneous multiwavelength four-dimensional optical microscopy.
Carlton PM, Boulanger J, Kervrann C, Sibarita JB, Salamero J, Gordon-Messer S, Bressan D, Haber JE, Haase S, Shao L, Winoto L, Matsuda A, Kner P, Uzawa S, Gustafsson M, Kam Z, Agard DA, Sedat JW.
Proc Natl Acad Sci U S A. 2010 Aug 12 (PubMed)
- WASH and the Arp2/3 complex regulate endosome shape and trafficking
Duleh SN, Welch MD
Cytoskeleton Vol. 67, 193-206, 2010 (PubMed)
- Distinguishing direct from indirect roles for bicoid mRNA localization factors
Timothy T. Weil et. al
Development Vol. 137, 169-176, 2010 (PubMed)
- The complexity of phosphorylated H2AX foci formation and DNA repair assembly at DNA double-strand breaks
Nakamura AJ, Rao VA, Pommier Y, Bonner WM
Cell Cycle. Vol. 9(2):389-97, 2010 (PubMed)
- Measurement of replication structures at the nanometer scale using super-resolution light microscopy
D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, and M. C. Cardoso
Nucleic Acids Res. Vol. 38: e8. 2010 (PubMed)