Summary of Contents for UVITEC Cambridge ALLIANCE IRIS
- Page 1 ALLIANCE IRIS QUICK MANUAL UVITEC CAMBRIDGE 2024/2025 Written by Sacha SAPSFORD, 10/12/2024...
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Page 2: Table Of Contents
IRIS QUICK GUIDE TABLES OF CONTENT Hardware Operation ..........................3 Introduction ............................3 Warranty ............................. 3 Power Supply and Connectivity ......................4 Turning on and off system ........................4 Inserting PADS ............................ 5 UV PAD ............................6 CHEMI PAD ..........................8 WHITE/BLUE PAD ........................ -
Page 3: Hardware Operation
Introduction The Alliance imaging system is a scientific instrument designed capture chemiluminescence or fluorescence gel or blots images. The Alliance IRIS system uses the IRIS software to control image capture and optimisation for selected applications. A cooled scientific CCD camera is used to capture high resolution digital images of protein and DNA bands in gels and on membranes obtained by electrophoresis or Western blotting separation methods. -
Page 4: Power Supply And Connectivity
Power Supply USB 3.0 RJ45 HDMI SWITCH PLUG Turning on system Turning on the system Alliance IRIS requires only a main standard plug that is to be plugged into its rear end (Figure 1a). -
Page 5: Turning On And Off System
*For a first-time use, please insert the UV PAD inside the unit BEFORE turning on the device* Commenté [SS1]: Tristan: Add switch on here Figure 1. a. Main plug inserted at the rear of Alliance IRIS b. Ensure screen appears like above before activating PC c. Button to start Alliance IRIS PC To turn device on: 1. -
Page 6: Inserting Pads
4. Turn off device by switching off my switch (Figure 1a) Inserting PADS Alliance IRIS has introduced a new generation of PADS in the system. Depending on the configuration of the system, it can include the following PADS: 1. UV light emitting diode (LED) pad 2. - Page 7 1 – UV PAD Figure 3. UV tag, chip and arrow facing the Figure 4. UV table inside unit. Ensure marks inside of the system. Proceed to slide UV (A), (B) and (C) are aligned table into empty compartment To verify successful connection: 1.
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Page 8: Chemi Pad
Figure 5. a. Protection screen b. UV300nm pad recognised by system c. Positioning of protection screen on UV pad d. UV warning button activated (circled) with UV LED light on (blue light) We recommend not to remove the UV pad from its original position. If so, please switch of the system before remove the pad from the unit. - Page 9 2. Select ‘Light and Filters’ 3. Check ‘Chemi Pad’ appears in the Light column (Figure 6b.) Figure 6. a. Positioning of chemi pad in the system b. Chemi Pad recognised by system and software For applications, this table will cover both chemiluminescent and epi-fluorescent applications. To remove the pad, place your left and right hand slightly below the chemi pad and lift up before sliding the pad towards you (Figure 7.).
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Page 10: White/Blue Pad
Alliance IRIS make use a novel technology known as ChromaScan. Details on the mechanism and positioning of the ChromaScan will be eventually released with the full Alliance IRIS manual. In this QUICK GUIDE, a simple introduction to the insertion of the ChromaScan excitation light source and... - Page 11 EXCITATION LIGHT SOURCE Figure 9a. presents the excitation light source of the Chromascan for epi-fluorescent applications. Up to 8 of these individual light sources can be inserted on the mechanical wheel located within the front part of the dark room of IRIS (Figure 9b.) Figure 9.
- Page 12 7. To place other light sources, manually rotate the mechanical wheel to spaces that are empty and repeat steps 4-6. 8. When complete, press ‘rescan’ on the software and check all the capsules have been recognised (Figure 11.) Figure 10. Insertion of excitation light source on mechanical wheel a. Orientation of ChromaScan excitation source b. Insertion of right side of the light source into the right-hand side (RHS) of the mechanical wheel c.
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Page 13: Emission Filters
*Do not travel with the excitation sources inserted inside the unit* EMISSION FILTER Alliance IRIS has integrated emission filters to its unit that are smaller and magnetic, facilitating the insertion or removal of these narrow bandwidth filters. If the need to add filters is required, please follow the following guidelines: 1. - Page 14 Commenté [SS4]: Tristan: Place each image below advice rather than altogether. Figure 12. Insertion of emission filters. a. Settings access b. Location of filter wheel door on right hand side of camera c. standard emission filter d. insertion of emission filter using right hand e. final result f.
- Page 15 To remove a filter, please follow the following: 1. Turn on system and software 2. Access software settings and select filter (Figure 12a) 3. Identify the filter you desire to remove and select ‘replace filter’ (Figure 12a) 4. Please wait for camera to lower before opening door 5.
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Page 16: Software Operation
SOFTWARE OPERATION Introduction The Alliance IRIS software is a licence-free software for image acquisition and analysis. The current version of the IRIS Software contains only the image acquisition capacity (2024). Hence, in this quick manual only this section will be explained. -
Page 17: System Settings
6. Once the process is over, flat fielding enhancement has been completed SERVICE The Alliance IRIS has its own system diagnostics. In the unlikely event the user identifies a bug or hardware malfunction, select ‘Park system’ in the service section of the settings. The system will lower its camera and proceed to diagnose the matter at hand. -
Page 18: Focus
Figure 15. Systems settings. Select tab desired, in this case, flat field tab was selected. Focus In order to view, correct and position correctly your sample, the function focus is to be used. This feature allows you correct the following: 1. - Page 19 2. SAMPLE HEIGHT To modify the focus as a result of a change in the sample height (i.e., 96-well plate), changes can be made here. Alliance IRIS software’s allows thus far the maximum height of 4cm (Figure 17.) Figure 17. Focus – sample height. To select desired height of sample, select this choice...
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Page 20: Light Intensity
3. LIGHT INTENSITY To correct the intensity of the white light, intensity of light can be changed here. Intensity can be changed to: 10%, 25%, 50%, 75% and 100% Figure 18. Focus – light intensity. To select light intensity, select between the 5 options;... -
Page 21: Capture Your Image
*Please be sure to select the correct tray for the application that is to be used* 1. To select protocol, click on Application 2. Alliance IRIS provides two columns; Standard (pre-defined protocols) and Custom (personalised protocols) 3. When working with the following, please select: o Chemiluminescence: select protocol termed ‘luminescence’... -
Page 22: Edit Application Type
Figure 20. Application. Access to application provides you towards pre-defined and custom- made protocols Edit Application Type With Alliance IRIS, the capacity to create or edit protocols based on a pre-defined protocol can be done. To do so: 1. Select Application 2. - Page 23 ➢ General: overview on device specifications ➢ Focus: Focus values used in protocol Following changes, press ‘save’ and name protocol to your desired taste. Once saved, the protocol will automatically appear under the custom column. Figure 21. Editing Application. Tab ‘edit application’ provides access to intricacies of image, allowing users to adapt the methodology of the photo to their application as well adding additional features including the inclusion of the marker or coloured images.
- Page 24 7. The Alliance IRIS will provide you 3 photos: signal image, marker image and overlay image 8. Save images Figure 22. Marker Addition. a. Final overlay option – all 3 photos (signal, marker and overlay) can be accessed by clicking the following < > circled. b. To ensure marker has been successfully...
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Page 25: Acquisition Mode
Acquisition Mode Before taking an image, the acquisition mode, found on the main page of the software allows you to capture the image in 3 different ways: 1. Automatic: The Alliance system automatically calculates the optimum exposure time. 2. Manual: The Alliance system allows the user to select the desired exposure time o Time definition mode: User defines a desired time ranging from 1 minute/second (min/sec) to 60-mins o Time Scroll Mode: User gets a live-preview of sample over a duration maximum... -
Page 26: Sensitivity And Aperture
Figure 23. Exposure mode(s). a. Select between automatic, manual and serial. b. In serial, there are two options; incremental and repetitive. Incremental allows users to define an initial and final exposure time where the software will automatically calculate an increment time between defined times. Repetitive will take a number of photos at the same X exposition time provided Sensitivity and Aperture Whilst in most cases the sensitivity and aperture are automatically pre-defined in the... -
Page 27: Light And Filter
o Sensitivity 1: High Sensitivity (native binning) o Sensitivity 2: Super Sensitivity Commenté [SS5]: Tristan: Definition o Sensitivity 4: Binning 2x2 o Sensitivity 8: Binning 4x4 o Preview mode: Epi-white light For aperture, the lower the value the greater the amount of light is being welcomed in by the camera. -
Page 28: Area Of Interest
Chemiluminescence No Light Chemiluminescence UV gels UV 300nm Ethidium Bromide (F590) Blue wavelength C480nm F500/F550 Green wavelength C520nm F600 Red wavelength C640nm F700 NIR wavelength C690nm F750 IR wavelength C780nm F850 Figure 25. Light & Filters. Area of interest The feature, area of interest, allows the user to define an area on the test image to calculate the auto-exposure time required for the specific part selected. -
Page 29: Acquire The Image
To manipulate the area of interest desired, select the white squares on the outside of the box to align and define your area of interest. When contempt with the choice, press continue to acquire final image. Figure 26. Area of interest. When in automatic mode, the option to select an area of interest will appear on the top right-hand corner. - Page 30 camera cooling and resolution can impact the final value – impacting 1) contrast or brightness of the image 2) the capacity to quantify correctly. To access the amount of grey scale captured, a bar on the left-hand side along with the image percentage (%) and display histogram are provided (Figure 27.).
- Page 31 Figure 27. Grey Scale Display. On the left hand-side the grey scale bar (1) can be readjusted by moving the two red squares along the bar. By selecting the image display percentage (%) (3), a representation of the % of shades of grey captured appears. To access the histogram and visualise the intensity of all grey levels selected, display (4) feature is provided whilst info (2) indicates the intensity of light of a single pixel at a certain region of interest marked out by the marker...
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Page 32: Save And Print
Save and print To save the photo, select the option ‘save’ located on the top left-hand side of the software (Figure 28.). When saving there are different formats to select from: o 16-bit tagged image file format (*.tif) – recommended for analysis o Open platform 16-bit tagged image file format (*.tif) –... -
Page 33: Visualise Your Image
Visualise your image: Gallery The Gallery function, located on the top left-hand corner, saves automatically the first 250 photos taken (whether it has been saved or not). Once it reaches 250, it will start deleting the first initial photo taken on the device. Figure 29. -
Page 34: Glp
Figure 30. Coloured image. Via paint motif, blot can be coloured based on personal preference To access information regarding the image, select the symbol (i) to view application and image protocol. All these information is automatically saved in the image and final report in analysis. Figure 31. -
Page 35: Inverse And Saturation
Inverse and Saturation In order to change the background of the image from black to white or white to black, the inverse function can be used. View figure 32 (1). To indicate saturation (pixel intensity > 65, 635 grey levels), select the button in figure 32 (2). Saturation is indicated in pink. - Page 36 Figure 33. Notes and 3D view. a. Notes tab to include comments on protocol b. 3D view of sample based on grey scale...
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