Addendum 2005-09-22 - GE Healthcare Life Sciences
Contents
1. Comments It is optional to add comments to your data To add comments to the data 1 On the bottom of the Setup Area click Experiment Comments to open the Experiment Comments dialog 2 Type in the Membrane ID the Individual Source and the Sample ID 3 Type in Comments for the scan To use the same comments for all the gels in this scan select Use this Comment for all Gels This option only applies to the Comments field All other fields are always specific to each gel area 4 Click Close To start the scan and for more information see 3 4 2 Start a scan and following chapters in the User Manual 28 4047 23 Addendum to Ettan DIGE Imager User Manual Edition AB2. 2 28 4047 23 Addendum to Ettan DIGE Imager User Manual Edition AB Filter Details Scanning Membranes Exposure 0 020 v 0 175 v Experiment Comments Em Fitter nim 680 30 Membrane O DIGE File Naming Format 595 25 Ex Filter ntm 540 25 635 30 w Pixel size pim 100 w Matrix type v v cY3 cys Dye In the Setup Area of the Setup View the parameters and settings for the scan are Gel Format User Select Chemistry Unassigned Channel v 1 2 Os O4 User Name GE Healthcare set membrane boundaries The scan area is shown as a blue box in the Grid Area The Gel format list is used to set the scan area In the Gel Format list in the Setup Area choose User Select In this mode you can manually set the area to be scanned Then use the mouse and drag a box from the positions of the The steps below do not have to be performed in a strict order Fig 2 Parameters and settings in the Setup Area Choose membrane from the Matrix Type list To set up a scan Matrix type 1 Gel format Setup EI RTS IS a IS AZ IS ES zie aga aa SUPE UE TCE Dr fe ie ee ji bon oo tot tot i 0 4 de ES dente qee OS as donde T oe ee ind EA tee teh i o sig ris Fasten re apa qero ara Ao an on ero are a wt A a Ce A A ee LU ER AT A AA OA AA A A ig A A A UA NE A O e E e po a tar Ri eo doriopedoje E Pad
3. Addendum 2005 09 22 Scanning Membranes 1 Loading a membrane When scanning membranes the cassette with permanently mounted glass plate Mounted Glass Cassette should be used To achieve optimal calibration when scanning the black plastic hold down plate should be placed on top of the membranes Note Wear powder free gloves when handling the cassette and membranes The powder used in laboratory gloves can fluoresce and may also scatter light affecting image quality To load a cassette 1 28 4047 23 Addendum to Ettan DIGE Imager User Manual Edition AB Clean the glass plate with distilled water and dry it using a lint free cloth Since fluorescent material has come into direct contact with the glass plate we recommend using a lint free tissue moistened with 10 hydrogen peroxide to remove this material followed by cleaning with distilled water Place the membrane s directly on the plate with the blotting side facing down If the membranels is wet make sure to exclude air bubbles when positioning It Note the positions of the membrane boundaries on the plate You will use this information to determine the area s to be scanned see 2 Set Up a membrane scan Place the hold down plate in the cassette on top of the membranes Put the lid on the cassette see Fig 3 4 in the User Manual and close the cassette by turning the locking cams up Insert the cassette into the scanner as described in 3 2 3 Inserting a cassette i
4. IR pay A A A O A AA ARA Dredd Poet AAA A A A A id A pint po Ji Ee A A A a IDODWLOT gt 3Z 3ZO0OEMr2 gt 2 gt x gt mM Fig 3 Grid Area 28 4047 23 Addendum to Ettan DIGE Imager User Manual Edition AB Scanning Membranes 2 Chemistry When scanning membranes the chemistry should be Unassigned 3 Pixel Size Select the pixel size i e resolution to use in the Pixel size list 100 um should be used 4 Channels and dyes Select the number of channels to be scanned by clicking the Channel check box Between one and four channels can be programmed Selection of the Channel check boxes must be performed in sequential numerical order For each selected channel select a dye to be scanned in that channel When selecting dye the excitation and emission filter to be used for that dye are automatically presented by the software The excitation and emission filter combinations have been selected to give the optimum results with minimal crosstalk Channel Dye Ex Fitter nm Em Fitter nm Exposure M1 cys 540025 595 25 0 020 v 2 cvs 63580 680 30 0 175 w Os vs v 0 400 v Ds O Ev User Name GE Healthcare Fig 4 Filter settings 5 Exposure Select the exposure to be used for each channel in the Exposure list Generally low exposures are used for membranes However a quick test scan on a small area should be performed to identify an optimal exposure See 3 4 1 Finding the optimal settings in the User Manual 6
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6. n the User Manual Scanning Membranes 2 Set Up a membrane scan Start the Scanner Software by double clicking the Ettan DIGE Imager icon J on the workstation desktop or by using the start menu Start Programs GE Healthcare Ettan DIGE Imager The scanner software Ettan DIGE Imager window opens in Setup View Eran DIGE imager He tet SeAFeme feat irreal Hen 1 The Too Bar E g 2 EQUINA fen Pepe 2 The Navigation Bar Bers Unease Gal Format Liner Select Peni scr qari Chemie Unassigned Miro Bye eran LL back Fie Naming Formal Channel Cha Er Fiber Fe Fiber Exposure Fine Detia H E 1 E HOR TAA pom Ectimided biim bra min eti Estimated Dish Space Needed ME Total Deck Space Ballads E Ea DATE mE 4 The Setup Area A toes OEGE ee 3 The Information Area ar e CTE oe A E E D E F a M i diat Khe L M Hi H P 5 The Grid Area PERE el H Fig 1 The Ettan DIGE Imager Setup View 1 The Tool Bar is used to manage scan templates and to turn on the scanner light 2 The Navigation Bar shows the status of the scan workflow 3 The Information Area shows the instrument status displays information about the scan and includes the SCAN button that is used to start a scan 4 The Setup Area contains the parameters and settings for the scan 5 The Grid Area defines the area or areas to scan The grid coordinates are aligned with those on the cassette
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