Multi-Mode Imaging of Membranes, Gels and Plates

Please acknowledge the SIP core facility ( RRID: SCR_018986) in publications, on posters, or in talks if you use any instruments in the SIP core facility. Please include SIP's RRID (RRID: SCR_018986) and the grant numbers for instruments funded through instrumentation grants in your acknowledgements. This is a requirement from the funding agencies and is crucial for future funding. Find example text on theÌýAcknowledgement PageÌýor the individual instrument pages.

Multi-Mode Imagingin Biochemistry, Biophysics and Structural Biology

The detection of biomolecules on gels, blots, and arrays is a foundational technique in biochemistry and biophysics, crucial for the separation, identification, and quantification of proteins, nucleic acids, and other macromolecules. These methods provide key insights into complex biomolecular processes including molecular interactions, cellular signalling pathways, and post-translational modifications, deepening our understanding of biological mechanisms in both health and disease.Ìý

The integration of multimode imaging, utilising multi-colour fluorescence, chemiluminescence, and colourimetric detection, significantly enhances analytical sensitivity, specificity, and dynamic range. Through multiplexing, researchers can combine these imaging techniques to analyse multiple molecular targets within a single experiment, adjusting detection methods and colour schemes to optimise sensitivity and dynamic range based on experimental requirements. This approach maximises data acquisition from a single sample, improving both the depth and accuracy of molecular analysis

Amersham Typhoon 5

Typhoon 5

The Typhoon 5 is a multi-mode, high-sensitivity imaging platform to capture high-resolution images from a multitude of samples. Ìý

Applications

The Typhoon has a wide range of applications including imaging of phospho storage screens (filmless autoradiography ), gel sandwiches, agarose and polyacrylamide gels, membranes, microplates, microarrays and even tissue slices.Ìý

Imaging Modes

The Typhoon 5 has 4 imaging modes that can be combined automatically. Fluorescence (5 channels), Phosphorimaging, Densitometry, and Chemiluminescence (dark scan)Ìý

Fluorescence Channels:

It has blue (488 nm), green (532 nm), red (635 nm), near IR (685nm) and far IR (785nm) excitation wavelengths and a wide choice of emission filters (IP,Ìý P 390BP, Cy2 525BP20, Cy3 570BP20, Cy5 670BP30, IRshort 720BP20, IRlong 825BP30, Cy2 LPB515, Cy3 LPG550, Cy5 LPR660) enable imaging of an extensive variety of fluorophores.Ìý

Automated multi-colour fluorescence scanning allows multiplexing of multiple targets in the same sample.

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Amersham IQ 800

IQ 800 Imager

The ImageQuant 800 imager from Cytiva/Fujifilm is equipped with a highly sensitive 8.3-megapixel, Peltier-cooled CCD detector and an F 0.74 lens, enabling high-resolution imaging of closely spaced bands. While it excels in the straightforward imaging of western blots and gels, it is versatile enough to support a wide range of additional applications.

Applications

The IQ 800 is capable of colour imaging a wide range of samples including imaging for various samples, including gels, membrane blots, multiwell plates, and petri dishes.

Imaging Modes

The IQ 800 has three imaging modes that can be combine automaticaly. Modes include chemiluminescence, fluorescence (5 channels), and colorimetric imaging.Ìý

Light Sources:

UV Epi 365 nm, Blue Epi 460 nm, Green Epi 535 nm, Red Epi 635 nm, IRshort Epi 660 nm, IRlong Epi 775 nm, White Epi 470 to 635 nm, White Trans 470 to 635 nm

Emission Filters:

Cy2 525 BP20 nm, Cy3(UV) 605 BP40 nm, Cy5 705 BP40 nm, IRshort 715 BP30 nm, IRlong 836 BP46 nm

Automated Signal-To-Noise Optimisation

The SNOW (Signal-to-Noise Optimisation Watch) imaging mode provides automated optimisation of parameters to detect faint bands in Western blot while ensuring image quality. SNOW increases sensitivity and extends dynamic range by preventing signal saturation. IQ 800 automatically stops once the optimal S/N is reached, removing the need for iterative manual adjustment of exposure and capture settings.

Essential Information for Using SIP's Multimode Imaging Platforms

Why?

Acknowledgements are essential for ensuring the continued success of the Shared Instruments Pool (SIP). They enable us to secure the necessary funding to sustain and expand the SIP, ensuring that our instruments are in optimal working condition and that the methods we offer are at the forefront of biochemical and biophysical research.

Please include SIP's RRID number (RRID: SCR_018986) in your acknowledgements. This allows funding organisations and potential grant reviewers to easily locate publications supported by SIP, helping to evaluate the impact of SIP on our research community.

If facility staff have provided substantial assistance, please consider acknowledging them. If they contributed significantly to the intellectual aspects or conducted important experiments, co-authorship may also be appropriate.

Example text:

  • We thank the Shared Instruments Pool (RRID: SCR_018986), Department of Biochemistry, University of Colorado Boulder for the use of the Amersham Typhoon 5 imager. The Typhoon 5 was funded by NIH S10OD034218.Ìý
  • We thank the Shared Instruments Pool (RRID: SCR_018986), Department of Biochemistry, University of Colorado Boulder for the use of the Cytiva IQ 800 imager.Ìý

Interested users can contact Dr Erbse to discuss planned experiments and arrange a training session. Training will be done using actual user samples, enabling users to collect preliminary data during the training and receive help from core staff right away if troubleshooting or optimisation is needed. Users are welcome to request additional training or support at any time. We are always happy to provide a refresher if it has been a while.

JSCBB users can sign up for imager time using the university's EMS scheduling system. Step-by-step instructions on how to book time through EMS can be found hereÌýEMS instructions

  • Consultation and Training is free of charge. SIP staff are happy to assist with a short pilot experiment if it can be accommodated within SIP's resources.
  • Regular user groups are expected to buy into SIP with a monthly flat fee according to their SIP usage level. For detailed information, please contact Dr Erbse.
  • Users are required to provide all consumables specific to their experiments.
  • Costs for necessary repairs, services, or replacement parts not covered by the service contract and due to normal wear and tear will be shared among all user groups, based on the time used over the past two years. Please note that assuming the instrument is handled properly, such repairs or replacements are infrequent, and costs may arise after a user’s period of use has ended.
  • Users are responsible for covering the costs of repairs or replacement parts needed due to damage caused by carelessness or neglect.

The both imaging platforms are located on the third floor of JSCBB in the C-Wing, room C380, on East Campus. Proxcard access is required at all times.