THE Autoradiography PRODUCT GUIDE. Hyperfilm. Kodak Film. Autoradiography Accessories

Transcription

1 THE Autoradiography PRODUCT GUIDE Hyperfilm Kodak Film Autoradiography Accessories

2 Introduction Amersham Biosciences is one of the world s leading suppliers of autoradiography and light detection products. Using its expertise in signal generation, Amersham Biosciences has developed Hyperfilm and accessory products specifically for use in life science applications, providing quality results and capturing the maximum amount of information. As well as our own well-established autoradiography range, the company is also a worldwide distributor of Kodak Scientific Imaging Systems autoradiography products. Amersham Biosciences is therefore able to advise customers on the best solution for their imaging needs. In addition to this catalogue, Review No.23, Efficient Detection of Bio-molecules by Autoradiography, Fluorography or Chemiluminesence, by R. Laskey, is also available. Review 23 is an independently written reference, that describes in detail, the principles of indirect and direct autoradiography. Review 23 is available on request, please contact your local sales office. Alternatively you can download a copy from the Web here. 1.

3 Contents Life science application guide 3 4 Autoradiography films: Hyperfilm MP 5 6 Hyperfilm ECL 7 8 Kodak BioMax MR 9 10 Kodak BioMax MS 11 Kodak BioMax Light 12 Kodak X-Omat AR (XAR) 13 Kodak X-Omat LS (XLS) 13 Autoradiography cassettes and screens: Hyperscreen Intensifying Screens 14 Kodak BioMax MS Intensifying Screens 15 Kodak BioMax TranScreen Intensifying Screens 16 Hypercassette Autoradiography Cassettes 17 Autoradiography accessories: Amplify Fluorographic Reagent 18 Darkroom Lamp 19 Hypertorch 19 ECL Mini-Camera 20 Hypercoat Nuclear Emulsions 21 Hyperprocessor Automatic Film Processor 22 Liquid Scintillation Cocktails Microscale Autoradiography Standards 25 Sensitize Pre-flash Unit 26 Processing Hanger 27 TrackerTape 27 Other film products: Kodak Duplicating Film 28 Kodak Micro-Autoradiography & TEM products 29 2.

4 Life science APPLICATION GUIDE Autoradiography methods and materials for life science applications Application Result required Label Southern/Northern blots high speed and sensitivity 32P Plaque/colony lifts accurate quantitation maximum speed and sensitivity optimum resolution non-radioactive, quantification Slot blots high speed and sensitivity 32P accurate quantitation maximum speed and sensitivity non-radioactive, quantification Dideoxy sequencing high speed 32P optimum resolution and maximum speed for difficult sequences good resolution and speed for routine work Cycle sequencing optimum resolution and maximum 33P speed for difficult sequences good resolution and speed for routine work 32P 33P/ 35 S ECL, Gene Images 32P ECL, Gene Images Protein synthesis, lysates maximum speed 35S/ 14 C/ 3 H optimum resolution Western blots maximum speed and sensitivity 125I accurate quantification optimum resolution 35S/ 14 C non-radioactive/maximum sensitivity ECL In situ hybridization macroscale in situ optimization 32P of parameters Receptor studies localization at cellular level: intermediate resolution 33P/ 35 S high resolution 33P/ 35 S localization subcellular level: high resolution 125I/ 3 H Whole body autoradiography high resolution 35S/ 14 C 35S 35S 33P 125I 3.

5 Method Film Support products Pre-flash and screens at -70ºC Hyperfilm MP Sensitize, Hyperscreen Screens at -70ºC BioMax MS BioMax TranScreen HE Direct autoradiography BioMax MR Pre-flash and direct detection Hyperfilm ECL Sensitize Pre-flash and screens at -70ºC Hyperfilm MP Sensitize, Hyperscreen Screens at -70ºC BioMax MS BioMax TranScreen HE Pre-flash and direct detection Hyperfilm ECL Sensitize Direct autoradiography Hyperfilm MP/BioMax MR Direct autoradiography BioMax MR Direct autoradiography Hyperfilm MP Direct autoradiography BioMax MR Direct autoradiography Hyperfilm MP Pre-flash and fluorography at -70ºC Hyperfilm MP Amplify, Sensitize Indirect autoradiography BioMax MS BioMax TranScreen HE Pre-flash and screens at -70ºC Hyperfilm MP Sensitize, Hyperscreen Screens at -70ºC BioMax MS BioMax TranScreen HE Direct autoradiography BioMax MR Pre-flash and direct detection Hyperfilm ECL Sensitize Direct autoradiography Hyperfilm MP Direct autoradiogaphy Micro autoradiography BioMax MR Hypercoat emulsion LM-1 Micro autoradiography Direct autoradiography Hypercoat emulsion EM-1 BioMax MR 4.

6 NEW FORMULATION Hyperfilm MP The new, higher performance photographic emulsion utilized in new formulation Hyperfilm MP not only offers up to 15% greater sensitivity over original Hyperfilm MP, but also improved contrast and resolution. Like its predecessor, new formulation Hyperfilm MP is an excellent multipurpose film constructed from a clear base coated on both sides with a photographic emulsion that is then protected by an anti-scratch layer. It may be processed using automatic processors or in tanks and trays using most common laboratory X-ray film developers and fixers. In a multi-discipline laboratory, performing both blots and sequencing experiments, new formulation Hyperfilm MP is the recommended film for high performance in all applications. New formulation Hyperfilm MP is also available in an envelope sealed format. Here, individual sheets of Hyperfilm MP are sealed in light tight envelopes giving maximum protection and ease of use in the life science laboratory. Each envelope is made from paper with an internal lining of low-density polymer. A single sheet of paper interleaving between the film and envelope prevents static screen 1 emulsion 2000 antiscratch layer plastic base emulsion Volume (AU) 1000 New Formulation Hyperfilm Current Formulation radioactive emissions light emission 32 P/ 125 I FIG 1 Diagram of Hyperfilm MP being used with an intensifying screen pg of target DNA FIG 2 Measure made on Personal Densitometer SI, analysis using ImageQuant. λ Hind III dots hybridized at 65 C in Church Buffer using 32 P Megaprime labelled probe at approx 1 ng/ml in hybridization buffer. Exposure time 24hrs at -70 C with Hyperscreen 5.

7 Applications 32P, 33 P and 125 I Northern, Southern and Western blots 35S, 33 P and 32 P sequencing Fluorography of 3 H, 14 C and 35 S New formulation film up to 15% more sensitive than traditional Hyperfilm MP when utilised with an intensifying screen Coated with emulsion on both sides to ensure optimum sensitivity Multipurpose film for the high speed detection of 3 H, 14 C, 35 S, 33 P, 32 P and 125 I Clear base offers high contrast for easy reading of images High resolution results at an economical price Designed for use in direct and indirect autoradiography, the film of choice for multipurpose work A New Hyperfilm MP B Old formulation Hyperfilm MP FIG 3 Single copy gene detection using 32 P. Southern blots of human placental DNA Hind III digest (1, 0.5, 0.25 µg per lane). Detection with N-ras insert probe (1.5kb) labelled with 32 P-dCTP. Autoradiography at 70 C with Hyperscreen for 36 hours. Product Hyperfilm MP RPN6K 18x24cm 25 sheets RPN1675K 18x24cm 75 sheets RPN36K 18x43cm 25 sheets RPN2115K 24x30cm 25 sheets RPN7K 30x40cm 25 sheets RPN8K 35x43cm 25 sheets RPN30K 35x43cm 75 sheets RPN1676K 5x7in 25 sheets RPN1677K 8x10in 25 sheets RPN1678K 8x10in 75 sheets RPN34K 20cmx25m 1 roll Hyperfilm MP envelope RPN6L 18x24cm 50 sheets RPN8L 35x43cm 50 sheets RPN1677L 8x10in 50 sheets 6.

8 NEW FORMULATION Hyperfilm ECL The new, high performance emulsion in new formulation Hyperfilm ECL delivers up to a 30% improvement in sensitivity over traditional Hyperfilm ECL with both blue and green light chemiluminescent systems.the emulsion of this double-coated film provides improved clarity and sensitivity for the detection of chemiluminescent-labelled samples, making it suitable for ECL, ECL Plus, CDP-Star and all other chemiluminescent systems. Film CD New Film Old Film Relative Blue LIght Emittance (Log Scale) Applications Chemiluminescent Northern, Southern and Western blots FIG 4 Comparison of luminescent response between new and old formulation Hyperfilm ECL A New formulation Hyperfilm ECL B Old formulation Hyperfilm ECL FIG 5 Western blots of post synaptic density preparation detected on new and old formulation Hyperfilm ECL for 30 second exposures. Primary antibody 1/2000 mouse anti-synapsin 1a/1b monoclonal, secondary antibody 1/1000 rabbit anti-mouse. Results supplied by Catherine Scott. Research School of Biosciences, University of Kent at Canterbury 7.

9 New formulation film up to 30% more sensitive than traditional Hyperfilm ECL High speed detection of chemiluminescence signal from nucleic acid and protein blots System tested with ECL and Gene Images kits to ensure optimum performance Clear base offers high contrast for easy reading of images Can be processed in automatic processors or manually using most common X-ray film developers and fixers Single coated emulsion, making it the film of choice for maximum resolution Clear base offers high contrast against blackened image for easy reading and scanning Product Hyperfilm ECL RPN2103K 18x24cm 25 sheets RPN3103K 18x24cm 75 sheets RPN2104K 30x40cm 25 sheets RPN1674K 5x7in 25 sheets RPN2114K 8x10in 25 sheets RPN3114K 8x10in 75 sheets RPN1681K 10x12in 25 sheets 8.

10 KODAK BioMax MR Kodak BioMax MR (maximum resolution) film, is a single emulsion film that utilizes T-Grain technology. The single emulsion format increases resolution and background clarity, making it the ideal choice for sequencing. It can be processed manually or in an automatic processor. Standard emulsion 35 S Sample Globular silver halide crystals cover less surface area Large mass of crystals stop 35 S beta particles, reducing efficiency per particle in exposing crystals FIG 6 BioMax emulsion increases sensitivity to 35 S, 33 P and 14 C Tabular grain silver halide crystals are flat, covering more surface area increasing number of interactions between beta particles and crystals Because crystals are flat each beta particle can expose multiple crystals FIG 7 Thermo Sequenase cycle sequencing using Redivue Nucleotides ( 33 P). 2 hour exposure on Kodak BioMax MR film Applications 35S and 33 P sequencing 9.

11 High speed direct autoradiography of samples labelled with 14 C, 35 S and 33 P T-Grain emulsion gives two-fold improvement in sensitivity over Kodak X-Omat AR film for direct exposure of 14 C, 35 S and 33 P Single coated emulsion, making it the film of choice for maximum resolution Clear base offers high contrast against blackened image for easy reading and scanning BioMax MR-1 Film (non-interleaved packaging) V x10in 50 sheets V x7in 25 sheets V x43cm 50 sheets V x24cm 50 sheets V x43cm 50 sheets V x30cm 50 sheets V x40cm 50 sheets V x61cm 50 sheets BioMax MR-2 Film (Ready Pack each sheet individually wrapped) V x10in 50 sheets V x7in 25 sheets V x43cm 50 sheets 10.

12 KODAK BioMax MS Kodak BioMax MS (maximum sensitivity) is a double sided film that utilizes Kodak T-Grain emulsion technology to maximise interaction with beta particles (see Fig 6, page 9) and is designed for maximum performance with 32P and 125 I. When used with the BioMax MS or BioMax TranScreen HE intensifying screens to detect 32 P, BioMax MS film forms one of the fastest conventional autoradiography film and screen systems available.the emulsion is extremely light sensitive, in particular to the green light emissions of BioMax intensifying screens. Note:This film should only be exposed to a red Kodak GBX-2 safelight filter using a 7.5 or 15 watt bulb. It can be processed manually or in an automatic processor. Applications 32P Northern and Southern blots State of the art technology, the film of choice for maximum sensitivity Reduces 4-day exposures to 1-day and overnight exposures to 4 hours Detects 1dpm bands in 1day Can be processed in automatic processors or manually (using most common X-ray film developers and fixers) Transcreen HE screen/ BioMax MS film Generic screen/ X-Omat AR film Net optical density (total density gross fog) BioMax MS film/ms screen XAR film/screen BioMax Light and XLS film/screen Relative log activity, 32 P (dpm), 40 hour exposure dpm/band 100-hour exposure at -70ºC FIG 8 Response to 32 P FIG 9 Comparison of film sensitivity to 32 P BioMax MS-1 Film (non-interleaved packaging) V x10in 50 sheets V x7in 25 sheets V x43cm 50 sheets V x24cm 50 sheets V x30cm 50 sheets V x40cm 50 sheets BioMax MS-2 Film (Ready Pack each sheet individually wrapped) V x10in 50 sheets V x7in 25 sheets V x43cm 50 sheets 11.

13 KODAK BioMax Light A double coated emulsion film from Kodak for the detection of chemiluminescence. Applications Hyperscreen Suitable for the detection of chemiluminescence Can be processed in automatic processors or manually using most common X-ray film developers and fixers Chemiluminescent blots 64 pg 64 pg 64 pg 32 pg 32 pg 32 pg 16 pg 16 pg 16 pg 8 pg 8 pg 8 pg 4 pg 4 pg 4 pg A B C FIG 10 Comparison of BioMax Film exposure to chemiluminescence. Nucleic acid probe, Human β-actin cdna, exposure time 5 minutes. A Kodak BioMax Light B Kodak X-Omat AR C Kodak BioMax MS BioMax Light-1 Film (non-interleaved packaging) V x10in 50 sheets V x7in 25 sheets V x43cm 50 sheets V x24cm 50 sheets V x30cm 50 sheets V x40cm Discontinued.xAlternativexproductxRPN2104K. BioMax Light-2 Film (Ready Pack each sheet individually wrapped) V x10in 50 sheets V x7in 25 sheets 12.

14 KODAK X-Omat AR (XAR) Kodak X-Omat (XAR) is a general-purpose film for use with all commonly used isotopes. Coated with emulsion on both sides, it provides high sensitivity for both direct exposure autoradiography and exposures with the Hyperscreen, BioMax MS or TranScreen Intensifying Screens. Applications Southern, Northern and Western blotting A popular general-purpose film, compatible with 35S, 14 C, 33 P, 32 P and 125 I X-Omat AR-5 Film (alternate interleaved packaging) V x10in 50 sheets V x7in 25 sheets V x43cm 50 sheets V x24cm 50 sheets V x30cm 50 sheets V x40cm 50 sheets V cmx25m 1 roll V inx50ft x 1 roll X-Omat AR-2 Film (Ready Pack each sheet individually wrapped) V x10in 50 sheets V x7in 25 sheets V x43cm 50 sheets KODAK X-Omat LS (XLS) Kodak X-Omat LS (XLS) is an economical, blue tinted film for use with all commonly used isotopes. It provides significantly lower detection sensitivity than BioMax or XAR films resulting in longer exposure times. Applications Southern and Western blotting, and sequences giving strong signals Features andbenefits Reduces costs where sensitivity is not required Similar performance to other blue tinted films X-Omat LS-1 Film (non-interleaved packaging) 13. V x10in (20.3x25.4cm) 50 sheets V x43cm (14x17in) 50 sheets V x7in (13x18cm) 25 sheets V x24cm (7x9.5in) 50 sheets

15 Hyperscreen INTENSIFYING SCREENS Hyperscreen Intensifying Screens are economical high performance calcium tungstate screens for use with 32 P and 125 I.They can be used with pre-flashed film to give an increase in sensitivity while retaining a linear response that allows quantitation of results. Applications Southern, Northern and Western blots using 32 P and 125 I Hyperscreen For rapid results and high sensitivity when detecting 32 P and 125 I, in Southern, Northern and Western blots Light emission from calcium tungstate phosphor, fully compatible with Hyperfilm MP Use with Sensitize Pre-Flash Unit for even higher sensitivity and accurate quantification Fine grain screens give clear band resolution Available in a full range of sizes to match + ++ Hypercassette and Hyperfilm MP minus + plus Hyperscreen ++ plus Hyperscreen and pre-flashed with Sensitize Kodak Intensifying Screen Cleaner Specially formulated solution for dust and dirt removal and static reduction, recommended for all Kodak and Hyperscreen Intensifying Screens FIG I-labelled IgG slot blots doubling dilutions (100nCi to 1.56nCi) 2 hour exposure to Hyperfilm MP FIG 12 Intensifying Screen Cleaner and Antistatic Solution Hyperscreen RPN x24cm 1 pair RPN x30cm 1 pair RPN x40cm 1 pair RPN x43cm 1 pair RPN x43cm 1 pair RPN x40cm 1 pair RPN1668 5x7in 1 pair RPN1669 8x10in 1 pair RPN x12in 1 pair Kodak Intensifying Screen Cleaner and Antistatic Solution V oz 1 bottle 14.

16 KODAK BIOMAX MS Intensifying Screens Kodak BioMax MS Intensifying Screens are special screens that can be used with any autoradiography film to increase sensitivity. Maximum sensitivity is achieved when used with BioMax MS film. Doubles the speed of conventional film Uses conventional intensifying screen format When used with BioMax MS Film it will increase speed of detection four fold (compared to normal film screen combinations) Applications Southern, Northern and Western blots using 32 P and 125 I X-Omat AR film/screen BioMax MS film/screen BioMax MR Film BioMax MS Film and MS Screen 3 H 14 C 35S 33P 32 P 32P & 125 I Sample Overcoat (Includes Protective Dye) 7 mil Estar Base Intensifying Screen Emulsion Layer Emulsion Layer FIG 14 BioMax film and intensifying screen: maximum speed for 32 P blotting applications FIG 13 Principle of direct autoradiography and normal intensifying screens Before 72 hours NOW 16 hours 32P labelled slot blots exposed at -70 C Kodak BioMax MS Intensifying Screen V x7in 1 screen V x10in 1 screen V x24cm 1 screen V x43cm 1 screen V x30cm 1 screen V x40cm 1 screen BioMax MS-1 Film and Screen Combination Pack (non-interleaved, 50 sheets per box with BioMax MS Intensifying Screen) V x10in 1 each V x43cm 1 each 15.

17 KODAK BIOMAX TranScreen INTENSIFYING SCREENS BioMax TranScreen Intensifying Screens incorporate a new, innovative technology and unlike conventional screens they capture the beta particles prior to their being exposed to the film.the BioMax TranScreen then converts the energy from the beta particles to photons without attenuation and directs the photons to the film. TranScreen LE This is the first and only intensifying screen for 35 S, 14 C, 33P, 45 Ca, 59 Fe and even 3 H labelled samples. Improves the resolution of 32 P samples compared to traditional screens. TranScreen LE When used with BioMax MS Film it allows images of 35S, 14 C, 33 P, 45 Ca, 59 Fe and even 3 H to be detected at least 5 times faster than conventional film screen systems Offers two-fold better resolution for 32 P and 125 I images (compared to normal intensifying screens) TranScreen HE Increases sensitivity at least four-fold (compared to normal intensifying screens) Improved handling features and image quality (compared to BioMax MS Screens) TranScreen HE Innovative intensifying screen technology for 32 P, 125 I and high energy emitting radiolabels. Low Energy TranScreen LE High Energy TranScreen HE 3 H 14C 35 S 33 P 32P 32 P & 125 I Sample Phosphor Base 7 mil Estar Base BioMax MS Film Reflector Hypercassette Phoshor TranScreen Surface Folder { Reflector surface Active side of blot FIG 15 Configuration of film, screen and sample when in use. BioMax MS film FIG 16 Principle of TranScreen Intensifying Screens. Note better resolution of TranScreen LE Applications 3 H and 14 C thin layer chromatography Southern, Northern and Western blots Sequencing with 35 S and 33 P Kodak BioMax TranScreen LE V x10in 1 screen V x43cm 1 screen Kodak BioMax TranScreen HE V x10in 1 screen V x43cm 1 screen 16.

18 Hypercassette AUTORADIOGRAPHY CASSETTES Hypercassette Autoradiography Cassettes have been designed for the toughest demands of life science research. Deep cassettes are 1.5cm in depth allowing them to accommodate thicker samples. Light tight exposures at temperatures as low as 70 C Long life, chip proof surfaces Optimum resolution from a design that ensures maximum, even contact between film and sample Minimal contamination as the non-porous pad gives a wipe clean interior Colour coding of work possible, choice of Hypercassette colours from red, blue or neutral Record details on polycarbonate label and wipe clean after use FIG 17 Hypercassette Autoradiography Cassettes Hypercassette, neutral (standard) RPN x24cm 1 cassette RPN x30cm 1 cassette RPN x40cm 1 cassette RPN x43cm 1 cassette RPN x43cm 1 cassette RPN x40cm 1 cassette RPN x7in 1 cassette RPN x10in 1 cassette RPN x12in 1 cassette Hypercassette, blue (standard) RPN x24cm 1 cassette RPN x30cm 1 cassette RPN x43cm 1 cassette RPN x10in 1 cassette Hypercassette, red (standard) RPN x24cm 1 cassette RPN x30cm 1 cassette RPN x43cm 1 cassette RPN x10in 1 cassette Hypercassette, neutral (deep) RPN x24cm 1 cassette RPN x40cm 1 cassette RPN x10in 1 cassette 17.

19 Amplify FLUOROGRAPHIC REAGENT Amplify is a fast, convenient fluorographic reagent for 35 S, 14 C and 3 H labelled samples in gels. Increased sensitivity can be achieved in addition to considerably reducing exposure times of these weak beta emitters. Increased detection efficiency 1000 fold for 3 H 15 fold for 35 S and 14 C Less than 2 hours total preparation time Odourless, does not contain DMSO or other organic solvents 35 S + 3 H + Versatile, can be used with acrylamide or agarose gels with no change in protocol Fully compatible with Sensitize and Hyperfilm MP FIG 18 Autoradiographs showing the detection of 35 S- and 3 H-labelled proteins in acrylamide gels with (+) and without ( ) Amplify. Samples exposed to pre-flashed Hyperfilm MP emulsion emulsion plastic base anti-scratch layer 3 H/Auger gel 3 H/Auger A B FIG 19 Internal absorption of weak β-emissions within a sample (A) compared to gel impregnated with Amplify (B). With Amplify β-emissions are converted to light and efficiently recorded on film Amplify Fluorographic Reagent NAMP100 1 litre 1 pack 18.

20 DARKROOM lamp The Darkroom Lamp is a socket type metal lamp that provides direct illumination. It screws into either a wall or drop-cord socket and holds a 5.5 inch diameter filter and a 7.5 watt standard incandescent bulb.the GBX-2 Safelight Filter is compatible with all autoradiography films including the highly light sensitive BioMax MS film. A 7.5 or 15 watt bulb is recommended. Socket type metal lamp that provides direct illumination FIG 20 Darkroom Lamp Darkroom Lamp V cm (5 1/ 2 in) diameter filter 1 lamp Note: Bulb and filter not included with Darkroom Lamp Safelight Filter, GBX-2 V cm (5 1/ 2 in) diameter 1 filter Hypertorch Hypertorch is a red light darkroom torch. A red LED produces a safe wavelength of light for short exposures to film. It allows visibility in areas of the darkroom, where fixed safe lights cannot reach e.g. cupboards. Ideal for use with Hyperfilm, Kodak autoradiography films and emulsions. Local illumination, minimizes film background Red LED, compatible with most autoradiography films Portable, allows all areas of darkroom to be illuminated Spring switch prevents accidental exposures FIG 21 Hypertorch battery-powered, darkroom pen torch Hypertorch RPN torches 1 pack (includes 2 AAA batteries) 19.

21 ECL Mini-Camera The ECL Mini-Camera is a camera luminometer specifically designed for use with ECL Systems, in particular Western blots from mini gels. It allows the chemiluminescent signal to be captured on Polaroid film without the need of a darkroom. Instant results at the bench on Polaroid film Compact and portable Handles blots up to a size of 52x77mm Polaroid film 667 is recommended for routine work while 612 can be used if greater sensitivity is needed. For work that requires the highest sensitivity and resolution use Hyperfilm ECL and conventional methodology. Applications Chemiluminescent blots from mini gels FIG 23 ECL Mini-Camera FIG 22 Doubling dilution (31, 15.5, 7.8, 3.9ng) of mouse IgG detected with anti-mouse Ig HRP and ECL Plus. Exposed for 1 minute ECL Mini-Camera Code RPN2069 (Supplied with 5 sample boats) Polaroid film is not supplied with the ECL Mini-Camera Quantity 1 camera Replacement Sample Boats Code RPN2079 Quantity 50 boats 20.

22 Hypercoat NUCLEAR EMULSIONS Hypercoat EM-1 and Hypercoat LM-1 are readyto-use nuclear emulsions that can be used in the microautoradiography of radiolabels in tissues or cells. When using 3 H the low energy of this isotope gives results of the highest resolution best appreciated using Hypercoat EM-1 which produces grains of a fine size. Isotopes such as 14 C, 35 S, 33 P, 32 P and 125 I are best detected with Hypercoat LM-1 which has a slightly larger grain size optimized for use with higher energy emissions. Both emulsions come ready-to-use and do not require diluting, a process that can increase backgrounds. Store in the dark at 2-8 C. Compatible with Hypercoat dipping vessels, specially designed glass vessels that facilitate maximum slide coverage from small volumes of emulsions. Ready-to-use, simply melt before dipping and coating High resolution, uniform crystal size and aggregate free Maximum sensitivity, high silver content allows reduced exposure times (0.09g Ag/g emulsion) Crystal sizes to suit all applications 0.2µm for general microautoradiography (Hypercoat LM-1) 0.13µm for high resolution light or electron microscopy of 3 H (Hypercoat EM-1) Applications Microautoradiography of tissues and cells FIG 24 Binding of [ 125 I]endothelin-1 to artery wall of patient with ischaemic heart disease, visualized with Hypercoat LM-1 Photograph courtesy of Dr M Dashwood, Department of Physiology, Royal Free Hospital School of Medicine, London FIG 25 Hypercoat Nuclear Emulsions and Hypercoat Dipping Vessel Product Hypercoat LM-1 RPN40 2x20ml 1 pack Hypercoat EM-1 RPN41 2x20ml 1 pack Hypercoat Dipping Vessels RPN39 1 pack 3 vessels 21.

23 Hyperprocessor AUTOMATIC FILM PROCESSOR Hyperprocessor is an advanced automatic film processor of compact bench top design. It is designed to give consistent high quality autoradiographs simply and quickly with Hyperfilm MP, Hyperfilm ECL, BioMax, XAR, XLS and most standard X-ray films. Sensors measure the film throughput and replenish chemicals as needed. Chemicals are maintained at optimum temperatures for quality results. Simple to operate. Develops, fixes, washes and dries automatically in 2 1 /2 mins Reproducible results. Internal control systems ensure consistent high performance Compact design, requires less than 0.2m 2 floor space Versatile. Can handle film sizes from 10x10cm up to 35x43cm High throughput. Can process 70 films, size 24x30cm, per hour Applications Automatic rapid processing of autoradiography films FIG 26 Hyperprocessor Contents Component Quantity Trolley 1 Reservoirs 2 Small spares kit 1 Full operator manual 1 Full installation manual 1 Product Hyperprocessor RPN v 1 processor Hyperprocessor RPN1700A v 1 processor ADDITIONAL INFORMATION Note: Hyperprocessor is not available in certain countries. Please contact your local office for further details. 22.

24 LIQUID SCINTILLATION Cocktails A comprehensive range of liquid scintillation cocktails compatible with most sample types. For aqueous samples: ACS - Organic scintillant. Efficiencies with 3 H of up to 50%, provided sample added is less than 6% ( v /v). Flashpoint 27.2 C. ACSII - Organic scintillant. Efficiencies with 3 H of up to 50%, provided sample added is less than 4.5% ( v /v). A flashpoint of 48.9 C reduces handling hazards. BCS - Biodegradable scintillant. Efficiencies with 3 H of 30-40%. Low odour, low toxicity and a high flashpoint (>150 C) makes it less hazardous to the user and the environment. PCS - Wide sample range phase-combining scintillant. Efficiencies with 3 H of 30-50%. Flashpoint 27.2 C. For non-aqueous samples: BCS-NA - Biodegradable scintillant. Efficiencies with 3 H of up to 45%. Low odour, low toxicity and a high flashpoint (>150 C) makes it less hazardous to the user and the environment. OCS High efficiency xylene-based scintillant. Efficiency with 3 H of >50%. Flashpoint 26.7 C. Tissue solubilizer: NCS Rapid and efficient solubilizer for biological materials and polyacrylamide gels prior to liquid scintillation counting. Compatible with BCS-NA and OCS scintillation cocktails FIG 27 Liquid Scintillation Cocktail specifications Product* Code Pack Size ACS Scintillation Cocktail NACS104 4x4litres ACS-II Scintillation Cocktail NACS204 4x4litres BCS Scintillation Cocktail NBCS104 4x4litres PCS Scintillation Cocktail NPCS104 4x4litres BCS-NA Scintillation Cocktail NBCS204 4x4litres OCS Scintillation Cocktail NOCS104 4x4litres NCS Tissue Solubilizer NNCS ml Product Code Pack Size [ 14 C] Scintillation Standard, N-hexadecane CFR6 75kBq, 2.0mCi, 1x10 6 dpm/g [ 14 C] Scintillation Standard,Toluene, CFR8 75kBq, 2.0mCi, 1x10 6 dpm/g [ 3 H] Scintillation Standard, N-hexadecane, TRR6 260kBq, 7.0mCi, 4x10 6 dpm/g [ 3 H] Scintillation Standard,Toluene TRR8 260kBq, 7.0mCi, 4x10 6 dpm/g [ 3 H] Scintillation Standard,Water TRR9 333kBq, 9.0mCi, 4x10 6 dpm/g [ 14 C] Sample Oxidiser Standard CFR dpm [ 3 H] Sample Oxidiser Standard TRR dpm 23. *ADDITIONAL INFORMATION Scintillation cocktails are not available in certain countries, please contact your local office for availability.

25 Selection guide 24.

26 Microscale AUTORADIOGRAPHY STANDARDS Microscale Standards are reference sources for autoradiography.the 3 H, 14 C or 125 I activity has been arranged in eight or ten layers of polymer, to give a good working range. Safe and easy to use as the radioisotope exists as a stable polymer Activity is uniformly incorporated into the polymer Arranged in 8 to 10 layers of increasing specific activity, to provide a good working range Applications Whole body autoradiography Receptor localization and receptor binding [ 3 H]Microscales [ 14 C]Microscales [ 125 I]Microscales RPA506 RPA507 RPA504 RPA511 RPA523 Bq/mg nci/mg Bq/mg nci/mg kbq/g nci/g Bq/g nci/g kbq/mg nci/mg FIG 28 Typical polymer activity Product Autoradiographic RPA kBq/mg, 3 110nCi/mg 20 strips (19mmx3.5mmx50µm) [ 3 H] Microscale RPA Bq/mg, nCi/mg 20 strips (19mmx3.5mmx50µm) RPA strips RPA strips RPA strips Autoradiographic RPA kBq/g, nCi/g 3 strips (19mmx3.5mmx120µm) [ 14 C] Microscale RPA504L kBq/g, nCi/g 20 strips (19mmx3.5mmx120µm) RPA Bq/g, nCi/g 3 strips (21mmx3.5mmx120µm) RPA511L Bq/g, nCi/g 20 strips (21mmx3.5mmx120µm) Autoradiographic RPA kBq/mg, nCi/mg 1 block (25mmx4.5mmx25µm) [ 125 I] Microscale RPA kBq/mg, nCi/mg 3 strips (25mmx4.5mmx20µm) RPA523L kBq/mg, nCi/mg 20 strips (25mmx4.5mmx20µm) 25.

27 Sensitize PREFLASH UNIT Sensitize is a specially designed flashgun for use with Hyperfilm ECL or Hyperfilm MP in conjunction with Hyperscreen Intensifying Screens to improve sensitivity and quantitation. It improves the linear range and increases film sensitivity. Most effective when detecting small amounts of radioactivity and chemiluminescent signal Calibration protocol ensures reproducibility High quality orange filter for optimum wavelength Sealed unit guards against white light leakage, often a problem with a conventional flashgun kb FIG 30 Sensitize Pre-flash Unit FIG 29 λ Hind III digests (10-fold dilutions) ECL detected and exposed to Hyperfilm ECL. Note the presence of 0.56kb fragment after preflashing Sensitize Preflash Unit Code RPN2051 Quantity 1 unit 26.

28 PROCESSING Hanger Clip-type metal film hanger for manual deep tank development of sheet films. Design ensures optimum film tension Accurate alignment of pin hole clamps permits easier washing near clips FIG 31 Processing Hanger Processing Hangers V x7in 1 hanger V x10in 1 hanger V x43cm 1 hanger TrackerTape TrackerTape is a phosphorescent tape which, when written on, can be used to label samples and corresponding autoradiographs. Applications Southern, Western and Northern blots Produces a permanent, negative image on autoradiographs Avoids confusion by allowing samples to be matched to results Can be used with ordinary marker pen Quick to use, image is produced in minutes FIG 32 Western blot identified by TrackerTape TrackerTape VRPN x15cm 10 sheets 27. ADDITIONAL INFORMATION Note: Not available in the USA.

29 KODAK Duplicating FILM A range of duplicating products which facilitate the generation of direct positive film transparencies of electrophoresis gels. Features andbenefits RA Duplicating Film Slower film for enhanced image quality and film stability Applications RA Duplicating Film single-emulsion film on a clear base that enables same-size reproductions of autoradiographs FIG P labelled blot reproduced on RA Duplicating Film Product RA Duplicating Film V x10in 100 sheets V x35cm 100 sheets V x43cm 100 sheets 28.

30 KODAK micro-autoradiography & TEM PRODUCTS A specially engineered range of micro-autoradiography products that facilitate high-quality images in Transmission Electron Micrography (TEM) and electron diffraction studies. Each of these products offers considerable latitude in performance, but they are best suited for medium-energy electrons (40 to 100kV), although acceptable results can also be obtained at higher electron-energy levels. Suitable for medium energy electrons (40-100kV) Range offers flexibility to meet most user needs Films have gelatin overcoat to minimise abrasion and a clean gelatin backing to reduce curl Kodak Electron Microscope Film 4489 This film has approximately half the speed of SO-163 film but exhibits less curl and shorter pump-down times. Kodak Electron Image Film SO-163 Approximately two times faster than 4489 film, however it requires longer pump-down times and shows greater film curl. When using SO-163 film for the first time, recalibration of the microscope exposure device is recommended. FIG 34 The Kodak TEM range Electron Microscope Film 4489 V x 4in 100 sheets V x 4in 250 sheets V x 9cm 100 sheets V x 9cm 250 sheets V x 11.9cm 100 sheets V x 7.0cm 100 sheets Electron Image Film SO-163 V x 4in 100 sheets V x 4in 250 sheets V x 9.0cm 100 sheets 29.

31 Amplify, ECL, ECL Plus, GeneImages, Hypercassette, Hypercoat, Hyperfilm, Hyperfilm device, Hyperprocessor, Hyperscreen, Hypertorch, ImageQuant, Megaprime, Microscale, Personal Densitometer, Redivue, Sensitize, Thermo Sequenase and TrackerTape are trademarks of Amersham Biosciences Limited or its subsidiaries. Kodak, BioMax, TranScreen and X-Omat are trademarks of Eastman Kodak Company. Polaroid is a trademark of Polaroid (UK) Limited. CDP-Star is a trademark of Tropix Inc and protected under one or more US patents numbers 5,851,771; 5,538,847; 5,326,882; 5,145,772; 4,978,614; and 4,931,569. All goods and services are sold subject to the terms and conditions of sale of the company within the Amersham Biosciences group which supplies them. A copy of these terms and conditions is available on request. Amersham Biosciences UK Limited Amersham Place Little Chalfont Buckinghamshire England HP7 9NA Amersham Biosciences AB SE Uppsala Sweden Amersham Biosciences Inc 800 Centennial Avenue PO Box 1327 Piscataway NJ USA Amersham Biosciences Europe GmbH Munzinger Strasse 9 D Freiburg Germany Amersham Biosciences UK Limited All rights reserved.

32 For more information visit our website: Asia Pacific Tel: Fax: Denmark Tel: Fax: Latin America Tel: Fax: South East Asia Tel: Fax: Australasia Tel: Fax: Finland & Baltics Tel: +358-(0) Fax: +358-(0) Middle East and Africa Tel: +30 (1) Fax: +30 (1) Spain Tel: Fax: Austria Tel: Fax: France Tel: Fax: Netherlands Tel: Fax: Sweden Tel: Fax: Belgium Tel: Fax: Germany Tel: Fax: Norway Tel: Fax: Switzerland Tel: Fax: Canada Tel: Fax: Italy Tel: Fax: Portugal Tel: Fax: UK Tel: Fax: Central East & S.E. Europe Tel: Fax: Japan Tel: Fax: Russia & other C.I.S./N.I.S. Tel: +7 (095) Fax: +7 (095) USA Tel: Fax: