Core Biotechnology Services

Olympus IX81 - Inscoper

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This Olympus IX81 microscope system was originally funded by the Wolfson Foundation and installed in autumn 2007 as a combined ScanR/CellR system and was mainly used for high content screening station. A second system, Nikon microscope 3 in the MSB, was upgraded with high content screening software in 2013, funded by the University Equipment Fund.

In 2022 this system was upgraded to Windows 10 with BBSRC funding as part of the Midlands Open Bioimaging (MOB) initiative. The upgrade included a new LED light source, new excitation and emission filters, new camera and a new PC. The system was also upgraded with new acquisition software from Inscoper.

Olympus objectives

There are a list of objectives available for the Olympus imaging systems.

See the Olympus objectives

This system can handle slides, 35 mm dishes and multi well plates. A slide holder for 4 slides is available.

This microscope is build around an inverted IX81 motorized microscope running Inscoper acquisition software. This system is equipped with an Hamamatsu ORCA Flash 4.0 LT+ camera (pixel size 6.5x6.5 µm and 82 % peak quantum efficiency), DIC objectives, hardware ZDC (Zero Drift Compensation) and software autofocus, linear encoders for high xy-precision, CO2 and a stage incubator for live cell imaging.

The Inscoper software allows for imaging of fixed samples or live cells and includes a high-content screening option. This makes it possible to screen both large and small numbers of samples. It has fully automated image acquisition which allows the user to select the number of images per well, the number of wells and the different filter sets. Results can be analysed using the the powerful Olympus ScanR image analysis software for automated object detection and cell separation using histograms and gating settings to include or exclude data sets.

Current software: Inscoper 7.1.15

LEDs:

365, 435, 470, 500, 550, 580, 635nm

Filter wheel with excitation filters

  • DAPI (376/30)
  • CFP (430/24)
  • GFP/FITC(480/30)
  • YFP (500/20)
  • mRFP/TRITC (555/25)
  • TxRed (560/40)
  • Cy5 (640/30)

Emission filter cubes on the microscope

  • DAPI/FITC/TRITC/Cy5; 418 - 450 nm, 505 - 532 nm, 577 - 610 nm, 664.5 - 725 nm (Chroma 89402m)
  • CFP/YFP; 455 - 484 nm, 520 - 554 nm (Chroma 29017m)
  • GFP/RFP;  
  • HqFITC; 510 - 565 nm
  • Cy5 (Dichroic 660LP, emission 700/75m)

Image analysis

Data acquired on this system during high content screening can be converted for compatibility with ScanR analysis software. 

Publications

Swaih AM, Breda C, Sathyasaikumar KV, Allcock N, Collier MEW, Mason RP, Feasby A, Herrera F, Outeiro TF, Schwarcz R, Repici M, Giorgini F (2022) Kynurenine 3-Monooxygenase Interacts with Huntingtin at the Outer Mitochondrial Membrane. Biomedicines. 10:2294

Elgamoudi BA, Ketley JM (2018) Lighting up my life: a LOV-based fluorescent reporter for Campylobacter jejuni. Res Microbiol. 169:108-114.

Zondler L, Miller-Fleming L, Repici M, Gonçalves S, Tenreiro S, Rosado-Ramos R, Betzer C, Straatman KR, Jensen PH, Giorgini F, Outeiro TF (2014) DJ-1 interactions with α-synuclein attenuate aggregation and cellular toxicity in models of Parkinson's disease. Cell Death Dis. 5, e1350; doi:10.1038/cddis.2014.307

Meinke P, Mattioli E, Haque F, Antoku S, Columbaro M, Straatman KR, Worman HJ, Gundersen GG, Lattanzi G, Wehnert M and Shackleton S (2014) Muscular Dystrophy-Associated SUN1 and SUN2 Variants Disrupt Nuclear-Cytoskeletal Connections and Myonuclear Organization. PLoS Genet. 10(9):e1004605.

Repici M, Straatman KR, Balduccio N, Enguita FJ, Outeiro TF and Giorgini F (2013) Parkinson’s disease-associated mutations in DJ-1 modulate its dimerization in living cells. J. Mol. Med. 91:599-611.

Mammalian SUN protein networks at the inner nuclear membrane and their role in laminopathy disease processes (2010) Haque, F., Mazzeo, D., Patel, J.T., Smallwood, D.T. Ellis, J.A., Shannahan, C.M. and Shackleton, S. J Biol Chem. 285: 3487-3498.

Automated high throughput mapping of promoter-enhancer interactions in zebrafish embryos (2009) Gehrig, J., Reischl, M., Kalmar, E., Ferg, M., Hadzhiev, Y., Zaucker, A., Song, C., Schindler, S., Liebel U. and Müller, F. Nature Methods 6: 911 - 916.

SIP1 protein protects cells from DNA damage-induced apoptosis and has independent prognostic value in bladder cancer (2009) Sayan, A.E., Griffiths, T.R., Pal, R., Browne, G.B., Ruddick, A., Yagci, T., Edwards, R., Mayer, N.J., Qazi, H., Goyal, S., Fernandez, S., Straatman, K., Jones, G.D.D., Bowman, K.J., Colquhoun, A., Mellon, J.K., Kriajevska, M., and Tulchinsky, E.PNAS 106:14884-14889.

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