Olympus Cell^R/scan^R system

Location: Adrian Building G1.
Contact: Dr Kees Straatman (krs5/7085, 2263).

If you export images out of the CellR software the pixel size information will be lost.

ScanR-acquisition manual
ScanR-analysis manual
ScanR-analysis 2.2 update
Read application note
Xcellence manual

In the Adrian Building three Olympus microscopes, funded by the Wolfson Foundation, were installed in autumn 2007. In April 2008 this microscope suite was named 'The Wolfson Foundation light microscopy facility'. Our live cell imaging and screening station is one of the microscopes in this facility. A second system, Nikon microscope 3 in the MSB, has been upgraded with screening software funded by the University Equipment Fund in autumn 2013.

Available objectives

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

Scan^R/Cell^R System

This microscope is an inverted IX81 motorized microscope which combines the Scan^R screening platform and Cell^R imaging station in one system. The Scan^R screening platform can be used for assay development and high-content screening using multiwell plates as well as single slides. 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. Many brands of multiwell plates are preprogrammed. Results can be analyzed with the powerful image analysis software for automated object detection and cell separation using histograms and gating settings to include or exclude data sets. For live cell imaging as well as for imaging of fixed materials the Cell^R option can be used. This allows fast acquisition and multiple position imaging for many hours. Protocols can be written in the Cell^R software to do complicated image and filter sequences.

This system is equipped with a Hamamatsu ORCA-AG CCD camera (pixel size 6.45x6.45 µm), DIC objectives, hardware ZDC (Zero Drift Compensation) and software autofocus, linear encoders for high xy-precision, CO2 and a stage incubator for live imaging.

In June 2013 we updated the CellR software to the new Xcellence software, version 1.2. This software replaces the CellR software but has basically the same look and feel. The ScanR Acquisition and ScanR analysis software have been updated to version 2.4.0.13 for Acquisition and 2.4.0.11 for Analysis.

Filter wheel with excitation filters for:
DAPI (350/50)
CFP (430/25)
GFP(470/22)
FITC (492/18)
YFP (500/20)
mRFP (556/20)
TxRed (572/23)
Cy5 (640/30).

Emission filter cubes on the microscope:
DAPI/FITC/TRITC (U-61002bs, U-61002m)
CFP/YFP
GFP/RFP (U-51019x, U-51019bs, U-51019m)
HqFITC (Dichroic 505LP, emission 535/50)
Cy5 (Dichroic 660LP, emission 700/75m)

We have a second licence for both the Scan^R analysis software and the Cell^R software to analyze data on the analysis computers in room G1a.

Some publications using the Scan^R screening station:

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.
Reverse transfection on cell arrays for high content screening microscopy (2007) Erfle H, Neumann B, Liebel U, Rogers P, Held M, Walter T, Ellenberg J, Pepperkok R. Nat Protoc.;2(2):392-9.
An RNAi screening platform to identify secretion machinery in mammalian cells (2007) Simpson JC, Cetin C, Erfle H, Joggerst B, Liebel U, Ellenberg J, Pepperkok R. J Biotechnol. 30;129(2):352-65.
High-throughput RNAi screening by time-lapse imaging of live human cells(2006) Neumann B, Held1 M, Liebel1 U, Erfle1 H, Rogers1 P, Pepperkok R, Ellenberg J. Nature Methods 3, 385-390.
High-throughput fluorescence microscopy for systems biology (2006) Pepperkok R, Ellenberg J. Nat. Rev. Mol. Cell Biol.;7(9):690-6