The Odyssey Fc Imager is the first CCD imager that offers excellent infrared (IR) fluorescent and chemiluminescent imaging performance in one system. The Odyssey Fc optical system combines solid-state laser diodes at 685 and 785 nm with a low-noise, CCD camera for detection of IR and chemiluminescent signals.
Chemiluminescence detection is simplified with the Odyssey Fc Imaging System. Film and darkroom steps are eliminated, along with the hazardous waste and excessive water usage of film development.

FieldBrite™ XT technology acquires images over the full dynamic range every time, with minimal user adjustments. With this new advance in CCD imaging, high performance is achieved without supercooling of the camera. A patented filtering technology guarantees low total system noise for acquisition times up to one hour. As the acquisition progresses, signal-to-noise ratio is optimized to provide a superior image.
The Odyssey Fc System provides 22-bit images, which deliver more than six logs of linear dynamic range. This allows accurate capture of both strong and weak signals on the same image.
FieldBrite XT offers:

• No need to supercool camera to reduce background noise
• One button acquisition for optimal image, with little or no user intervention
• Uniform imaging over entire field of view
• Increased image acquisition time results in improved signal to noise

With the proprietary technologies of the Odyssey Fc Imaging System, a single instrument can perform quantitative IR fluorescent detection PLUS qualitative chemiluminescent detection. Publication-quality images are easy to produce with either detection method. (Because chemiluminescence is a time-dependent enzymatic reaction, its quantitative range is limited. IR fluorescence is superior for quantification.)

The Odyssey Fc Imaging System includes LI-COR Image Studio Software for fast, easy image acquisition and analysis. At the push of a button, targets of interest can be imaged over more than six logs of linear dynamic range in every acquisition.

Overview and Advantages:

• Wide Linear Dynamic Range - Accurately detect strong and weak signals over a broad linear dynamic range.
• Two-Color Detection and Quantification - Detect two targets simultaneously on the same membrane increases the accuracy of quantification and comparison.
• High Sensitivity - Near-infrared fluorescence provides sensitivity equal or better than chemiluminescence giving you low background, high signal-to-noise, multiplexed detection, and the sensitivity of any fluorescent system.
• Image Studio Software - User-friendly software with easy navigation makes acquisition and analysis straight-forward.
• Direct Detection for Better Data and a Cleaner Planet - No film or darkroom needed. IRDye signals on membranes are stable indefinitely if stored properly.
• Flexible System with Many Applications - The Odyssey Fc system has been used for many applications, including Western blot analysis, EMSA, In-Gel Westerns, Coomassie gel documentation and DNA gel documentation.
• Buy from a Leader - LI-COR Biosciences is the leader in fluorescent Western blot detection and near-infrared imaging.

Applications:

Western Blotting

Quantitative IR Western Blotting and Chemiluminescent Detection
IR fluorescent detection is a direct, nonenzymatic approach to Western blotting that uses secondary antibodies labeled with IRDye® infrared dyes. This method improves quantitative accuracy and reproducibility, and facilitates multiplexing. The stable fluorescent signal can be detected immediately, or after extended storage.
Qualitative Western blotting detection with various chemiluminescent substrates can also be done using the Odyssey Fc.

[ABOVE] The Odyssey Fc System can acquire images in both fluorescent and chemiluminescent modes. IR Fluorescence: Lasers are used to excite 700 nm and 800 nm IRDye fluorophores for 2 channel detection. Fluorescent emission is then detected to generate an image of the sample. At these IR wavelengths, membrane autofluorescence is very low. This method directly detects the fluorescently labeled conjugates. Because the fluorescent signal is very stable, samples can be archived and re-imaged. Chemiluminescence: Luminol substrate is applied to the sample, and is oxidized by horseradish peroxidase (HRP) enzyme in a light-generating reaction. This method is indirect, because it relies on the enzymatic reaction to report the location of the HRP conjugate. Chemiluminescent signals are dynamic and time-dependent. Light emission is initially strong, but diminishes over time.

Molecular weight markers can be visualized at 700 nm
on chemiluminescent Western blots

[LEFT] NIH/3T3 cell lysates were separated and transferred to a membrane. Odyssey Protein MW Markers (P/N 928-40001, 928-40000) (which are labeled with 700 nm IR fluorescent dye) were included in the left lane. Akt was then detected, using appropriate antibodies and ECL Plus™ (GE Healthcare) chemiluminescent substrate.
Akt signal was acquired with the Odyssey Fc System in the chemiluminescence channel (white bands) and 700 nm fluorescence channel (MW marker; red bands). The two signals were then overlayed in Image Studio software, so that both signals could be visualized simultaneously.

Multiplex Westerns

The two infrared fluorescent detection channels of the Odyssey® Fc System enable simultaneous two-color analysis – an advantage that is not available with chemiluminescent or radioactive methods. Two primary antibodies are used, each from a different host species (e.g., rabbit and mouse). IRDye® secondary antibody conjugates (e.g., anti-rabbit and anti-mouse) are then used to detect the primary antibodies and visualize the targets.
Multiplex analysis makes normalization easy, and eliminates error introduced by stripping and reprobing or by comparison of separate blots. Superior image clarity and detail make it easier to detect subtle mobility shifts caused by protein modifications such as phosphorylation.
The Odyssey Fc System can detect Western blots in three ways:

• IR Fluorescence at 700nm
• IR Fluorescence at 800nm
• Chemiluminescence

[ABOVE] Lysates of EGF-treated A431 cells were separated and transferred to nitrocellulose. The blot was probed with anti-ERK and anti-phospho-ERK primary antibodies, and then detected with IRDye 680LT and IRDye 800CW secondary antibodies. Blot was imaged with Odyssey Fc System for 2 min. This phospho-ERK antibody crossreacts with phospho-EGFR (upper green band).

Membrane Autofluorescence and Multiplexing

Autofluorescent background can greatly influence detection sensitivity. Low background increases the signal-to-noise ratio, making faint signals detectable and extending the linear dynamic range. At the IR wavelengths used by the Odyssey Fc System, intrinsic membrane autofluorescence is extremely low. At traditional visible wavelengths, membrane autofluorescence is much higher and can mask weaker signals.
True multiplex Western blot detection requires excellent sensitivity in both fluorescent channels – and only IR fluorescence can achieve this sensitivity. Visible fluorescence typically provides acceptable detection sensitivity in only one fluorescent channel.

[ABOVE] Nitrocellulose and PVDF membranes were imaged at 700 and 800 nm wavelengths, then imaged at 532 and 635 nm (Cy3 and Cy5 channels) with a GenePix® 4100A (Molecular Devices). Membrane background was dramatically reduced at IR wavelengths

Gel Documentation

After electrophoretic separation of protein or nucleic acid samples, gels may be stained and imaged with the Odyssey Fc Imager.
IRDye® Blue (a colloidal Coomassie stain) is commonly used to stain and view polyacrylamide protein gels. Coomassie is a strong IR fluorophore, and can be imaged at 700 nm with the Odyssey® Fc System to achieve superior sensitivity.
Syto® 60, a near-infrared fluorescent DNA stain, can be imaged at 700 nm – and is more sensitive than ethidium bromide.

[ABOVE] Purified fetuin protein was serially diluted (9.1 – 0.5 µg) and separated on a 10% gel. The gel was stained with IRDye Blue and imaged with Odyssey Fc in the 700 nm channel. Fetuin resolved as a doublet (2 min acquisition).

[ABOVE] DNA samples (50 bp or 1 kb ladder) were pre-incubated with Syto 60 prior to gel loading. Samples were separated on a 1.2% agarose gel at 80V for 1 h. For each ladder, amounts loaded were 1, 0.5, 0.25, and 0.125 µg (left to right). Gel was imaged with Odyssey Fc in 700 nm channel (2 min acquisition).

Specifications: