GE Healthcare Life Sciences Data file 29-0044-13 AB Imaging systems, software, and accessories Typhoon™ FLA 9500 biomolecular imager Typhoon FLA 9500 (Fig 1) is a robust and versatile laser scanner that is ideally suited to multiuser environments. Biomolecular imaging applications include sensitive and quantitative measurements of Western blots, multiplex fluorescence, and radioisotopic labels by storage phosphor as well as digitization of colorimetric stains. Typhoon FLA 9500 delivers: • Versatility: imaging of multifluorescent-, chemifluorescent-, radioisotope-labeled, and colorimetric samples • High resolution and quantitation: a pixel resolution of up to 10 µm and a linear signal response over five orders of magnitude provides precise quantitation in gels, blots, tissue sections, and arrays • High sample throughput: scanning area of 40 × 46 cm enables simultaneous imaging of up to 20 gels or blots, measuring 10 × 8 cm in size. This facilitates comparisons among blots, and reduces workload and waiting time • Flexibility: optimized performance for new applications by adapting the system with stages, detectors, filters, and lasers Fig 1. Typhoon FLA 9500 is a high performance, versatile laser scanner for sensitive and quantitative measurements in a multiuser environment. The system provides several imaging modes such as fluorescence, filmless autoradiography, and digitization of colorimetrically stained gels. For chemiluminescence detection of low abundance proteins the ImageQuant™ imager series is recommended. The system is optimized for differential protein expression studies and quantitative protein detection, e.g. multifluorescent Amersham™ ECL Plex™ imaging for precise quantitation of two or more proteins in the same Western blot. • 2-D DIGE imaging: simultaneously image two 2-D DIGE gels for differential expression studies • Visible and infrared fluorescence imaging: optional near infrared excitation for imaging IRDye™ and other infrared dyes Typhoon FLA 9500 is a variable mode laser scanner with modular access to the optical components (Fig 2) and excitation sources, providing both versatile and flexible imaging for precise quantitation of proteins, nucleic acids, and other biomolecules (Table 1). Fig 2. Filters are easily exchanged by the user.

Broad linear dynamic range Table 1. Typhoon FLA 9500 specifications Typhoon FLA 9500 provides a broad linear dynamic range in all detection modes, for example when using Cy™5 labeled proteins (Fig 3). Detection modes: Fluorescence, phosphorimaging, digitization, and chemiluminescence Excitation wavelengths: 473 nm (blue LD laser), 532 nm (green SHG laser), 635 nm (red LD laser), 685 nm (optional near IR LD laser), and 785 nm (optional near IR LD laser) Radioisotopes: Dynamic range: Standard filters: Pixel sizes: Scanning area: Optional filters: Dimensions (W × H × D): 900 × 400 × 800 mm Weight:...

Table 2. Emission filters Wavelength range (nm) Detection examples BP390 Cy2, SYBR™ Green, FAM™, FITC, Alexa Fluor™ 488, SYPRO™ Ruby, SYPRO Orange, GFP Cy2 DIGE Fluor, ECL Plex Cy2 Lasers can be exchanged in the field to accommodate new applications and fluorophores. The system can house up to four lasers simultaneously, from a choice of five laser excitation wavelengths (473, 532, 635, 685, and 785 nm). Cy3 DIGE Fluor, ECL Plex Cy3 Cy5, Alexa Fluor 633, TOTO™ 3, DiD, Cy5 DIGE Fluor, ECL Plex Cy5 Carbonic anhydrase in LMW marker Amersham ECL Gel 4-20% SDS-PAGE tris-glycine Excitation Emission...

Carbonic anhydrase Cy3 and Cy5 (data not shown) labeled 81mer oligonucleotide TBE-Urea 1.0 mm Excitation Emission filter 532 nm BPG1 (570DF20) 635 nm LPR (665 LP) 0.2 fmol for both Cy3- and Cy5-labeled 81mer Sensitive multiplex detection of Western blots Typhoon FLA 9500 is a versatile scanner that is ideal for imaging of fluorescent Western Blot membranes. This method is very sensitive and the signal is proportional to protein quantity. Moreover, it is possible to detect more than one protein at the same time by means of secondary antibodies labeled with different fluorophores. Amersham ECL...

1 - Control: Cell lysate of human cell line A431 2 - Treated: Cell lysate of human cell line A431 treated with EGF 24 cm 3-11 NL Precast low-fluorescent DIGE gel Excitation Emission filter 473 nm BPB1 (530DF20) 532 nm BPG1 (570DF20) 635 nm DBR1 (665LP) Fig 9. Overlay image of a two-dimensional difference gel electrophoresis (2-D DIGE) gel with control and treated samples, and internal standard. The control and treated samples were labeled with Cy3 and Cy5 DIGE Fluors (minimal dye labeling protocol). The internal standard sample was labeled with Cy2 DIGE Fluor. The data sets were evaluated using...

Typhoon FLA 9500 has two PMT’s that are optimized for sensitive detection over a broad spectrum from visible light and up to the near infrared (NIR) region. Figure 13 shows how the multialkali PMT 2 gives a linear response and sensitive detection of an IRDye 680 labeled secondary antibody in a Western blot. Excitation light passes through the sample and excites a fluorescent plate. The emitted light from the plate passes through the sample again and is collected and converted to an electrical signal. The method is suitable for documentation of colorimetrically stained gels. Data are stored either...

Ordering information System Quantity Code number * Includes 473 nm, 532 nm, and 635 nm lasers, filter tray, IP filter, LPB filter, LPG filter, LPR filter LPR filter for ch2, BPB1 filter, BPG1 filter, Fluor Stage, Membrane Weight, Phosphor Stage, Low Fluorescent Glass Plate Stage, Multi Stage with TP plug-in. Fluorescent plate for digitization, capture software, USB cable, mains cables (EU and USA), User Manual, Getting Started Guide. One license of ImageQuant TL software is provided with Typhoon FLA 9500. Upgrades and accessories* Quantity Code number Phosphorimaging plate, Phosphorimaging plate,...