Page 1 ANDOR A USER’S GUIDE TO ELECTRON MULTIPLYING CCD Version 1A...
Page 3 CONTENTS...
Page 4: Table Of Contents
CONTENTS Section 1 ABOUT THE ANDOR iXon CCD Introduction to the Andor iXon CCD 1 . 1 Technical Support 1 . 2 Section 2 INSTALLATION Safety Precautions 2 . 1 The iXon CCD System 2 . 2 Installing the System 2 .
Page 5 CONTENTS WORKING WITH PROGRAMS Section 5 Introduction 5 . 1 Editing Programs 5 . 2 Running Programs 5 . 3 Section 6 THIRD PARTY SOFWARE Soft Imaging System 6 . 1 Other S/W supported 6 . 2 TUTORIALS Section 7 Tutorial 1 7 .
Page 6: Technical Support
ABOUT THE ANDOR iXON CCD Introduction 1 . 1 Technical Support 1 . 2 Section 1...
Page 7: Introduction
Please feel free to contact Andor Technology directly, or your local representative or supplier, if you have any questions regarding your iXon system. Andor Technology Ltd...
Page 8: Using The
INTRODUCTION Using The In the software, all the controls you need for an operation are grouped and sequenced Manual appropriately in on-screen windows. As far as possible, the descriptions in this User’s Guide are laid out in sections that mirror the Windows Interface. The following type-styles are used in the right-hand margin of the User’s Guide to pick out key features of the menus:- •...
Page 9: Technical Support
ANDOR TECHNOLOGY E-mail sales@andor-tech.com 435 Buckland Road, Rosewood Building, South Windsor, CT 06074, USA Tel. (860) 648-1085 Fax. (860) 648-1088 Japan ANDOR TECHNOLOGY JAPAN E-mail sales@andor-tech.com 503 Ichibancho Central Building 22-1 Ichiban-cho, Chiyoda-ku Tokyo 102-0082, Japan Tel. 81-3-3511 0659 Fax. 81-3-3511 0659...
Page 10: Safety Precautions
INSTALLATION Safety Precautions 2 . 1 The iXon CCD System 2 . 2 Installing the System 2 . 3 Using the iXon CCD 2 . 4 File Information 2 . 5 Hot Keys 2 . 6 Section 2...
Page 11 INSTALLATION …...
Page 12: Safety Precautions
SAFETY PRECAUTIONS Working with Electronics The computer equipment that is to be used to operate the iXon CCD Detector should be fitted with appropriate surge/EMI/RFI protection on all power lines. Dedicated power lines or line isolation may be required for some extremely noisy sites.
Page 13 Your detector is a precision scientific instrument containing fragile components. ALWAYS HANDLE THE DETECTOR WITH CARE. Additional statement regarding Equipment Operation If the equipment is used in a manner not specified by Andor Technology Ltd, the protection provided by the equipment may be impaired. Environmental Conditions...
Page 14 SAFETY PRECAUTIONS Electrical Ratings 5Vdc 20 Watts The external Power Supply Block will supply a maximum additional 7.5V at 30 Watts Section 2 . 1 Page 14...
Page 15: The Ixon Ccd System
THE IXON CCD SYSTEM The Andor iXon CCD system is composed of hardware (notably the detector head and the card), the software, and documentation (including on-line help, the User’s Guide to Andor iXon CCDs, and the Programmer’s Guide to Andor Basic). This section of the User’s Guide identifies the main components of the system and guides you through the installation procedure.
Page 16 THE IXON CCD SYSTEM Main Components of iXon CCD System Section 2 . 2 Page 16...
Page 17 THE IXON CCD SYSTEM The Detector Head The Detector Head contains the CCD Sensor and its Pre-Amplifier. It also contains the Temperature Sensor, the pre-amplifier for the temperature sensor, and the Thermoelectric Cooler. The head can be attached to a microscope or other optical device for acquiring data.
Page 18: Smb Connectors
THE IXON CCD SYSTEM Detector Head to Controller Card Cable The 3m Cable that connects the detector head to the Controller Card uses a proprietary 36-way connector. It is well shielded against electrical interference. Power Supply Block (PSB) See SECTION 4.12 SETTING THE TEMPERATURE - Cooling Performance and Operation for details of temperatures that can be achieved when using the supplied PSB.
Page 19 THE IXON CCD SYSTEM External Trigger Signal Fire, Arm and Shutter Signal Section 2 . 2 Page 19...
Page 20 THE IXON CCD SYSTEM Computer Requirements IBM AT Compatible Computers offer considerable computing power at relatively modest cost. The system requires a PCI-compatible computer. The PCI slot you use must have bus master capability. The minimum recommended specification is a 800 MHz Pentium, with 256 Mbytes of RAM, and a hard disk, and a CD-ROM for installation of the software.
Page 21: Installing The System
INSTALLING THE SYSTEM Installing the Controller Card Install the PCI Controller Card as you would other slot-in cards - such as graphics cards. Consult the manual supplied with your personal computer to ensure correct installation of the Controller Card for your particular PC. Perform the installation as follows: •...
Page 22 INSTALLING THE SYSTEM Connecting the System Connect the elements of your system as follows: • Plug your PC into the mains outlet to ensure grounding, but keep the power switched off. • Connect the Detector Head to the Controller Card using the Cable provided. The 36-way connectors on the cable are polarized so that there is only one way of installing the cable.
Page 23 INSTALLING THE SYSTEM • Insert the Andor CD. The ‘installation wizard’ now starts. (If it does not start automatically, run \setup.exe on the CD.) Follow the on-screen prompts. • Now restart the computer. • Run the Andor application: from the PC’s desktop select Start…Programs…Andor iXon …Andor iXon •...
Page 24: Using The Ixon Ccd
USING THE IXON CCD The Main Window is your ‘entry point’ to the system. The menu options that you select from the Main Window either execute functions directly, or launch further windows and dialog boxes that let you select the functionality you require.
Page 25 USING THE iXON CCD Main Window (Default) Main Window (Displaying Data) Section 2 . 4 Page 25...
Page 26 USING THE iXON CCD Main Window (Working with Programs) Section 2 . 4 Page 26...
Page 27: File Information
FILE INFORMATION If you have an active Data Window on screen, clicking the File Information button Information on the Main Window launches a File Information Window. The File Information Window displays file details and lets you enter your own notes. The Filename associated with the active Data Window.
Page 28 FILE INFORMATION File Information Dialog Box Section 2 . 5 Page 28...
Page 29: Hot Keys
HOT KEYS Hot keys (or shortcuts) let you work with the system directly from the keyboard, rather than via the mouse. F1 provides Help. The following hot keys relate to data acquisition: Key Strokes Description Take signal scan Autoscale Acquisition Ctrl + B Take background Ctrl + R...
Page 30 HOT KEYS Many of the hot keys relate to the Data Window. Key Strokes Description Display Mode Image • • • Expand (‘Stretch’) data-axis • • • Contract (‘Shrink’) data-axis If maintain aspect ratio off, • • • Expand (‘Stretch’) x-axis If maintain aspect ratio on, Expand (‘Stretch’) x-axis and y-axis If maintain aspect ratio off,...
Page 31 HOT KEYS Key Strokes Description Display Mode Image • • PgUp Scroll up through tracks Scroll down through tracks • • PgDn Shift + PgUp Move to next image in series • • • • • • Shift + PgDn Move to previous image in series Move cursor left •...
Page 32 HOT KEYS Key Strokes Description Display Mode Image • Peak search left Ctrl + Left Arrow Peak search right • Ctrl + Right Arrow • Toggle Palette • • • Reset • • • Rescale • • • Alt + F9 Toggle Rescale Mode •...
Page 33 INSIDE THE iXON CCD Section 3...
Page 34 INSIDE THE iXON CCD … Electron Multiplying Charge Coupled Devices 3 . 1 EMCCD Gain 3 . 2 Section 3...
Page 35: Electron Multiplying Ccds
ELECTRON MULTIPLYING CCDS Typical EMCCD Chip EMCCD Structure Specification (no./size of pixels etc.) varies according to the chip model Image area Storage area Output amplifier Shift Gain register register Advances in sensor technology have led to the development on a new generation of ultra-sensitive, low light CCDs.
Page 36: Emccd Gain
ELECTRON MULTIPLYING CCDS Impact Ionization One of the electrodes (phases) in the Gain Register is replaced with two electrodes, the first is held at a fixed potential and the second is clocked as normal, except that much higher voltages (between 40V and 60V amplitude) are used than are necessary for charge transfer alone.
Page 37 ELECTRON MULTIPLYING CCDS The shift register also has a series of electrodes (which are vertical, i.e. parallel to the columns) that are used to transfer the charge packets, one element at a time, into the output node of the ‘on-chip’ amplifier. The output of the amplifier feeds the analog-to-digital converter (A/D), which in turn converts each charge packet into a 14 / 16-bit binary number.
Page 38 ELECTRON MULTIPLYING CCDS Readout Sequence of an EMCCD Only subset of pixels shown! Amplifier Output Node Read Exposure to light causes a pattern of charge (an electronic image) to build up on the frame (or ‘image area’) of the EMCCD-chip. Section 3 .
Page 39 ELECTRON MULTIPLYING CCDS Charge in the frame is shifted vertically by one row, so that the bottom row of charge moves into the shift register. Charge in the shift register is moved horizontally by one pixel, so that charge on the endmost pixel of the shift register is moved into the Gain register, and subsequently into the output node of the amplifier.
Page 40 ELECTRON MULTIPLYING CCDS Vertical and Horizontal Binning Binning is a process that allows charge from two or more pixels to be combined on the EMCCD-chip prior to readout (see Readout Sequence of a EMCCD above.) Summing charge on the EMCCD, and doing a single readout gives better noise performance than reading out several pixels and then summing them in the computer memory.
Page 41 ELECTRON MULTIPLYING CCDS Vertical Binning of Two Rows Only subset of pixels shown! Gain Register Amplifier Read Output Node Exposure to light causes a pattern of charge (an electronic image) to build up on Section 3 . 1 Page 41...
Page 42 ELECTRON MULTIPLYING CCDS the frame (or ‘image area’) of the CCD-chip. Charge in the frame is shifted vertically by one row, so that the bottom row of charge moves down into the shift register. Charge in the frame is shifted vertically by a further row, so that the next row of charge moves down into the shift register, which now contains charge from two rows - i.e.
Page 43 ELECTRON MULTIPLYING CCDS In the following example, where each superpixel is of dimensions 2x2 pixels, charge from two rows is first binned vertically into the shift register; then charge from two pixels of the shift register is binned horizontally into the output node of the amplifier.
Page 44 ELECTRON MULTIPLYING CCDS Vertical & Horizontal Binning (2x2 Superpixels) Only subset of pixels shown! (… 5) Steps 1 - 4 as in Vertical Binning of Two Rows Amplifier Amplifier Output Output Node Node Read Charge from two rows has already been vertically binned into the shift register (see ‘Vertical Binning of Two Rows’, above).
Page 45 EMCCD GAIN Section 3. 2...
Page 46: Emccd Gain
EMCCD GAIN The output from the gain register is fed into a conventional CCD output amplifier. Gain and Noise This amplifier, even in a scientific CCD, will have a readout noise of a few electrons rms and around 10 or 20 electrons rms at MHz readout rates. However this noise will effectively be reduced by the multiplication factor of the gain register which, when high enough, will achieve noise levels below 1 electron rms.
Page 47 EMCCD GAIN Darksignal or dark current is just the same as in conventional CCDs. When judging The EMCCD and Darksignal the required operating temperatures to eliminate the darksignal contribution, usually a temperature where the darksignal shot noise is comfortably below the readout noise is selected.
Page 48 EMCCD GAIN The gain of an EMCCD system varies with temperature. The graph below shows Gain Temperature Dependence how the gain multiplication increases as the temperature decreases. Curves are shown for various software gain settings and the figures are typical values. So if a system is operated at room temperature it will have reduced gain.
Page 49 EMCCD GAIN This graph shows how the EM Gain setting on the software is related to the actual electron multiplication factor for various temperature settings. Again the figures are typical. 1000 -60C -50C -40C -30C -20C Gain Setting on Software (EM Gain) Note: At gain setting 0, the Electron Multiplication process is disabled, and amplification does not take place Section 3 .
Page 50: Working With Data
WORKING WITH THE SOFTWARE Working with Data 4 . 1 Getting Help 4 . 2 Handling Files 4 . 3 Data Acquisition 4 . 4 Setting The Acquisition 4 . 5 Choosing the Data Type 4 . 6 Displaying Data 4 .
Page 51 WORKING WITH THE SOFTWARE Section 4 Section 4...
Page 52: Working With Data
WORKING WITH DATA The rich functionality of the software package is described in detail in the remainder of the User’s Guide. However, if this is the first time you have used an Andor detector, the short descriptions that follow will help familiarize you with its design philosophy and some of its key terminology.
Page 53 WORKING WITH DATA You can open several Data Windows to look at the same data set (possibly in different levels of detail or in different Display Modes): each Data Window has the same name and #n (which identify the data set), but a unique number :x (following the data set name) to identify the window itself.
Page 54 WORKING WITH DATA The Command Line The Command Line gives you ready access to all functions and arithmetic data processing of the Andor Basic programming language - without the need to write programs! However, to process the contents of a data set, the data set must first be in memory (RAM), and a corresponding Data Window will therefore be on screen.
Page 55: Getting Help
GETTING HELP The software provides On-Line Help typical of Windows applications. For details of how to use On-Line Help, Consult your Windows User’s Guide. On-Line Help can generally be launched from a Help button on the window or dialog box about which you require help. Alternatively, press F1 on your keyboard. On-Line Help will provide you with information that is relevant to the part of the application from which help was called.
Page 56: Data Acquisition
DATA ACQUISITION File The File Menu makes available typical Windows facilities to let you Open and Save files, as well as facilities specific to Andor’s Software to let you create or run programs. For further details of Windows file handling dialog boxes (e.g. the Open and Save as dialog boxes) consult your Windows User’s Guide.
Page 57 DATA ACQUISITION This is faster and more reliable than making use of the virtual memory feature of the operating system (which simulates extra physical memory using a buffer on the hard drive to hold data larger than would normally fit in the RAM available) because Andor’s Virtual Memory system is specifically designed to handle the large file you are attempting to load and work with in the MCD application.
Page 58 DATA ACQUISITION Batch Conversion Batch Conversion allows conversion of multiple .sif files, stored on disk, into various formats. There are three main sections to the dialog: Source, File list and Destin Source To convert multiple files they must be in the same directory on disk. This directory is selected in the upper section of the dialog, either by typing in the path or by selecting the browse button and navigating to the appropriate directory.
Page 59 DATA ACQUISITION When the above selections have been made, the Convert button at the bottom of the dialog should be enabled (if not check that the source and destination directories are valid and that there is at least one file selected in the list of sifs). Pressing the convert button may bring up another dialog box depending on which conversion type was chosen.
Page 60 DATA ACQUISITION Display Mode Readout Export as… Options Mode TIFF ASCII JPEG MPEG Image Image 2D / 3D Exporting Data as ASCII text, means you can subsequently import your data into ASCII other applications (such as spreadsheets) that use the ASCII format. When you click the OK button on the ‘Export As’...
Page 61 DATA ACQUISITION Tagged (or Tag) Image File Format (TIFF, .tif) is used for storing bitmapped images TIFF and is widely supported by commercial publishing packages. When you save data as a .tif file, the system automatically creates a file containing a number of labels (tags), which describe key properties of the data (gray levels, byte format, etc.) As with BMP, the image is exported WYSIWYG fashion.
Page 62 DATA ACQUISITION Option saves a data set to a 32-bit integer .dat file. The limits for the 32 bit integer are handled in similar fashion to the 16 bit integer above. Option saves a data set to a 32-bit floating point .dat file. When using your own software to handle a .dat file, you have to work out how many bytes to read in.
Page 63: Print Setup
DATA ACQUISITION Run Program executes the contents of an active Program Editor Window (see Run Program SECTION 5.3 RUNNING PROGRAMS). Run Program by Filename launches an ‘Open’ Dialog Box. Select the file that Run Program by Filename contains the program you want to run. The program begins execution as soon as the file is selected (see SECTION 6.3 RUNNING PROGRAMS).
Page 64: Setup Acquisition
DATA ACQUISITION Preparing to Acquire Data Once you have installed the software and hardware, you are in fact able to acquire Acquisition data (as Counts) simply by clicking the Take Signal button on the Main Window. However, to help you prepare to acquire data, the Acquisition Menu on the Main Window provides the following options: Setup Acquisition launches a Setup Acquisition Dialog Box where you can select Setup Acquisition...
Page 65: Take Signal
DATA ACQUISITION If you select Autoscale Acquisition (the ‘tickable’ item on the Acquisition Menu) Autoscale Acquisition the system will configure the Acquisition Window, if necessary adjusting its scales in real time, so that all data values are displayed as they are acquired. The manner in which the data are displayed corresponds with the selection you make under Rescale Data Mode on the Display Menu, where you may choose to display values between zero and maximum (0...max);...
Page 66: Video Mode
DATA ACQUISITION The data you acquire using Take Reference are displayed as counts minus background under the Ref tab of the Acquisition Window. The data type you select via Setup Data Type on the Acquisition Menu may require you to perform Take Reference before you perform Take Signal. Note: that you do not necessarily have to take background data prior to each acquisition of reference data: if the data acquisition parameters remain unchanged since you last performed Take Background, then no new background data are...
Page 67: Setting The Acquisition
SETTING THE AQUISITION Setting the Acquisition The Setup Acquisition Dialog Box lets you determine the manner in which data are acquired. Launch it either from the Setup Acquisition option on the Acquisition Menu (see SECTION 4.4 DATA ACQUISITION - Preparing to Acquire Data), or by clicking the Setup Acquisition button The Setup Acquisition Dialog Box is divided into the following menus: Setup CCD...
Page 68: Acquisition Mode
SETTING THE AQUISITION Setup CCD Menu When you select the Setup Acquisition Dialog, the software will default to Setup CCD as the first Dialog menu. This menu will allow you to set the parameters for a number of areas including; Acquisition Mode, Triggering, Timings for Exposure and Kinetic Cycles, Electron Multiplying Gain, and Vertical Shift Speed.
Page 69: Single Scan
SETTING THE AQUISITION In Single Scan acquisition mode, the system performs one scan (or readout) of Single Scan the CCD. Single Scan Exposure Time e.g. Exposure Time = 0.3 s • Exposure Time in seconds - i.e. the time during which the CCD collects light prior to readout.
Page 70 SETTING THE AQUISITION The Accumulate acquisition mode allows you to add together in computer Accumulate memory, the data from a number of scans to create an ‘accumulated scan’. Accumulate Exposure Time e.g. Exposure Time = 0.3 s Accumulate Cycle Time = 1.5 s No.
Page 71 SETTING THE AQUISITION The Kinetic Series acquisition mode allows you to capture (into the same data set Kinetic Series - see SECTION 4.1 WORKING WITH DATA) a sequence of single scans. Kinetic Series Exposure Time e.g. Exposure Time = 0.3 s Kinetic Cycle Time = 3 s Kinetic Cycle Time No.
Page 72: Trigger Modes
SETTING THE AQUISITION The Trigger Mode drop-down list box on the CCD Setup Dialog Box offers the Trigger Modes following options : Selecting the Internal Trigger Mode means that, once you issue a data Internal acquisition command (see SECTION 4.4 DATA ACQUISITION - Making an Acquisition), the system determines when data acquisition begins.
Page 73 SETTING THE AQUISITION Note: If you have shutters connected, and are using an External Trigger, you must ensure that the shutter is open before the optical signal you want to measure occurs. Normally, when using External Trigger the system will only enable the Fast External triggering of the system after a complete Keep Clean Cycle has been performed.
Page 74 SETTING THE AQUISITION The Exposure Time (in seconds) is the time during which the EMCCD collects Exposure Time light prior to readout. The system will default to a minimum Exposure Time should you attempt to enter too low a value. The system allows you use the EMCCD-chip at the heart of the detector in a Readout number of ways to collect and read out data.
Page 75 SETTING THE AQUISITION You may select a Vertical Shift Speed (the speed with which charge is moved Vertical Shift Speed down the CCD-chip prior to readout) from a drop-down list box on the CCD Setup Acquisition Dialog Box. The Vertical Shift Speed you select will affect the minimum exposure and cycle times available.
Page 76 SETTING THE AQUISITION Binning The Binning Dialog Box is the adjacent Dialog box to Setup CCD. This Dialog box allows you to set and control various Binning Patterns, and define Sub Images of the iXON CCD. Binning Dialog Box for iXon 87 CCD (512 x 512 pixels) By default, taking an Acquisition supplies you with a count from each pixel on the Sub Image CCD, in effect allowing you to take a picture of the light pattern falling on the pixel...
Page 77 SETTING THE AQUISITION To read out data from a selected area, or Sub Image, of the EMCCD use the radio buttons to select the resolution, which you require. The software offers a choice of up to four defined sub images, depending on the format of your iXon CCD camera. These pre-defined sub images are located in the centre of the chip, but can be moved by clicking and dragging the Sub Image outline.
Page 78 SETTING THE AQUISITION Full Resolution Image Full Resolution Image Superpixels Superpixels and Sub-area and Sub-area Superpixel 2-D Pixel Matrix Sub-area or sub-image The software presents a selection of five of the most common binning patterns; 1x1, 2x2, 4x4, 8x8, and 16x16 pixels. For example, if you enter 4x4 binning the CCD-chip is notionally divided into a matrix of superpixels which each measure 4x4 pixels and provide a signal for readout.
Page 79: Auto Save
SETTING THE AQUISITION Auto Save The Auto-Save Dialog Box is accessed by clicking the Auto-Save tab, on the Setup Acquisition Dialog Box. This menu allows you to set parameters and controls for the auto saving of acquisition files, removing the worry of lost data and files.
Page 80 SETTING THE AQUISITION · Operator name (supplied by user) · Computer name · Camera type · Date · Time Any combination of these may be selected by activating the relevant tick box. Please Note: This function will only Auto Save single scan, accumulated, or kinetic images.
Page 81 SETTING THE AQUISITION Spooling Spooling a computer document or data file is the process of reading it and storing it in a buffer, normally on a hard disk, so it can be printed or otherwise processed at a more convenient time. The Andor MCD software allows you to spool acquisition data direct to the hard disk of your PC.
Page 82 SETTING THE AQUISITION Video Mode Video Mode is a useful acquisition mode for focusing of your iXon CCD, and for watching experimental events happening in real time. However, this mode will not allow you to save any of the acquired images or data, except for the last frame of the sequence.
Page 83 SETTING THE AQUISITION • Exposure Time in seconds - i.e. the time during which the EMCCD collects light prior to readout. The system will default to a minimum Exposure Time should you attempt to enter too low a value. • Delay in seconds - i.e.
Page 84 SETTING THE AQUISITION The horizontal and vertical Binning parameters determine the dimensions of any Binning superpixels you may choose to create. Full Resolution Image Full Resolution Image Superpixels and Sub-area Superpixels and Sub-area Superpixel 2-D Pixel Matrix Sub-area or sub-image The software presents a selection of five of the most common binning patterns;...
Page 85: Image Orientation
SETTING THE AQUISITION Image Orientation The final Dialog box in the Setup Acquisition Menu is Image Orientation, which has tick boxes that allow you to flip and rotate image data as they are acquired. Image Orientation Dialog Box The Image Orientation Dialog also contains ‘R’ images, which allow you to easily visualize the various orientation options available.
Page 86: Choosing The Data Type
CHOOSING THE DATA TYPE The Data Type Dialog Box, launched from the Setup Data Type option on the Acquisition Menu, allows you to select the type of data you want the system to compute and display whenever you perform Take Signal (see SECTION 4.4 DATA ACQUISITION).
Page 87 CHOOSING THE DATA TYPE The illustration below shows typical use of Background, Reference and Signal for computations such as %Absorptance or %Transmittance. Use of Background, Reference and Signal Example: Percentage Transmittance, Absorptance light source sample detector Background Andor CCD empty InstaSpec IV Reference Andor CCD...
Page 88 CHOOSING THE DATA TYPE If you perform an operation ‘out of sequence’, the system will remind you what to do by launching an Acquisition Error Dialog Box. Acquisition Error Dialog Box Provided you do not change the acquisition parameters, the scans you take for background and reference are automatically used for subsequent data acquisitions whenever you perform Take Signal.
Page 89 CHOOSING THE DATA TYPE Note: ‘Signal’, as used in the definitions of the calculations, refers to ‘raw’ data from the CCD and should not be confused with the possibly ‘processed’ data to be found under the Sig tab of the Data Window. See SECTION 4.7 DISPLAYING DATA.) The data types available are listed overleaf.
Page 90 CHOOSING THE DATA TYPE The data type Counts represents raw, digitized data (i.e. no calculations have been Counts performed on the data) from the CCD detector’s analog to digital (A/D) converter. Refer to the detailed specification accompanying your particular CCD detector for the number of electrons that correspond to 1 count.
Page 91 CHOOSING THE DATA TYPE Percentage Transmittance represents the light transmitted by an object. If %Transmittance Reference is the background corrected incident intensity, and Signal - Background the transmitted intensity (i.e. the intensity of light which has been transmitted through the material being examined), then: % Transmittance = 100 x (Signal - Background) / Reference Absorbance Units are a measure of light absorbed by an object (i.e.
Page 92 CHOOSING THE DATA TYPE Attenuation Attenuation (dB) is a measurement, in decibels, of light absorbed due to transmission through a material - decibels are often used to indicate light loss in fiber optic cables, for instance. If Reference is the background corrected incident intensity, and Signal - Background the transmitted intensity (i.e.
Page 93 CHOOSING THE DATA TYPE Data Type Dialog Box Section 4 . 6 Page 93...
Page 94 CHOOSING THE DATA TYPE Functionality for displaying and manipulating data is only available if a Data File has been opened, or if data have been newly acquired, or if you are using the Andor programming language to create a new window in which to display data. In Basic each case data are displayed in a Data Window.
Page 95 CHOOSING THE DATA TYPE The menu item Scale to Active is only available if you are in 2D Display Mode and have chosen to overlay a number of traces: see Overlay later in this section. For more details on data displays see The Data Window next. For more details on the use of x-, y-, and data-axes in data displays see Axes later in this section.
Page 96 CHOOSING THE DATA TYPE bottom of the Data Window you will see up to three tabs, labeled Sig (Signal), Bg (Background), and Ref (Reference) which (where appropriate) identify data acquired via the functions Take Signal, Take Background, and Take Reference respectively. Click on the appropriate tab to display the trace you require.
Page 97: Displaying Data
DISPLAYING DATA Change Display Mode The system can display data in a number of ways. The display modes may be Change Display Mode accessed from the Change Display Mode option on the Display Menu or from the corresponding buttons on the Main Window, once data have been acquired or retrieved.
Page 98 DISPLAYING DATA Sub Image Mode Clicking the Sub Image Mode button on the main task bar will launch a Dialog box, with a choice of three options; Full Image, Cancel, or Store. This tool allows you to click and drag a Sub Image on the #0 Window only. This Sub Image region will then be used in subsequent acquisitions.
Page 99 DISPLAYING DATA Data Window (Grayscale Image without side traces) A Palette Bar, running across the top of the window, always shows the full palette The Palette Bar of grays or colors available in each color mode. The palette is graded so that lower data values correspond to the darker tones to the left of the palette, and higher data values correspond to the brighter tones to the right of the palette.
Page 100 DISPLAYING DATA Clicking the Rescale button on the button bar of the Main Window, when a Data Rescale Window is open, causes the system to display against an appropriate data scale (and, in the case of Image Display Mode, in appropriate colors or grayscale tones) all data that falls within your selected range on the x-axis.
Page 101 DISPLAYING DATA The computer ‘rescales’ your display by default: the full available Data-Axis palette of colors/grays is correlated with the full range of data you have acquired. When it rescales in Image Display Mode, the computer finds the maximum (Max) and minimum (Min) data values, and scales the available number of colors to the range, where the Range = Max - Min.
Page 102 DISPLAYING DATA Increasing and Decreasing Contrast You can use the pairs of left-right arrows at either end of the Palette Bar to increase or decrease the displayed data range (i.e. ‘shrink’ or ‘stretch’ the scale on the palette) and thereby alter the contrast of the data shown in Image Display Mode. Increase Displayed Decrease Displayed Data Range...
Page 103 DISPLAYING DATA Now only a fraction of the original data range is represented by the total number of colors. Decreasing the displayed data range produces high contrast. By holding down the left-right arrows at either end of the Palette Bar you can alter the minimum and maximum values in the displayed data range independently of each other, and so adjust the contrast to the level you require.
Page 104 DISPLAYING DATA Slide Data Scale Left Sliding the scale to the left means that the displayed data range covers higher data values (the numbers on the scale increase). The display becomes darker. Darken Image Note: When you slide the scale, you will see the image flash periodically as the system performs the brightness adjustment.
Page 105 DISPLAYING DATA Brightness: Overview Data-Axis To make the data shown in Image Display Mode brighter, the computer uses the same number of colors (as it used for rescaling) Original Data Range to scale lower data values. Thus any data originally represented by darker colors are now represented by brighter colors.
Page 106 DISPLAYING DATA Each additional Data Window, which bears the same # number as the active Data Window used to create it, is further identified by an extension (:2, :3, etc.) to the title on the window’s title bar. The original Data Window adopts the extension :1. The original and the additional Data Window(s) let you see and manipulate the same underlying set of data: for example, making a new acquisition (see SECTION 4.4 DATA ACQUISITION - Making an Acquisition) while a ‘family’...
Page 107 DISPLAYING DATA expanded: the system adds extra area as necessary so that the zoomed image accurately represents the height to width ratio of the individual pixels on your CCD- chip. Note: To help the system zoom the area you require, draw the Zoom Box in similar proportions to the height and width of the image display.
Page 108 DISPLAYING DATA Display Preferences Dialog Box (Image Preferences) Section 4 . 7 Page 108...
Page 109 DISPLAYING DATA Clicking the Axis Setup option on the Display Menu launches an Axis Setup Axis Setup Dialog Box. If you select Axis Setup while you are viewing data in 2D or a 3D Display Mode, you will be able to enter, in text boxes, the maximum and minimum values you wish to appear on the x- and data-axes (the horizontal and vertical display axes respectively) of your Data Window.
Page 110 DISPLAYING DATA Axis Setup Dialog Box (for 2D and 3D Display Mode) The Data Histogram Dialog Box can be launched by clicking on the icon on the Data Histogram Main Window, or through the Display drop down menu. This tool allows you to plot a histogram, or graph, between the maximum and minimum data points in the displayed range.
Page 111 DISPLAYING DATA In 2D Display Mode the system displays data as an unlabeled trace. To label peaks automatically, click 2D with peak labeling. While working in labeled mode, you may label a peak manually by double clicking it. To remove a peak label, double 2D with Peak Labeling click the labeled peak.
Page 112 DISPLAYING DATA Data Window (2D Display Mode with Peak Labeling) / Display Controls To read off a data value, click on the trace to position the cursor on the point of interest (you may need to use the left and right arrow keys on your keyboard to position the cursor precisely).
Page 113 DISPLAYING DATA The left and right edges of the zoom box demarcate the range of values that, after zooming, will be shown across the full width of the x-axis; the top and bottom edges of the zoom box demarcate the range of values that will be shown over the full height of the data-axis.
Page 114 DISPLAYING DATA Overlay allows you to display up to nine 2D traces simultaneously in the same Data Overlay Window (see previously), on the same set of axes. The Data Window in which you intend to display the overlaid traces must be in 2D Display Mode.
Page 115 DISPLAYING DATA You may add up to eight overlays to your original data trace. The original data trace is always displayed in blue. Each overlay appears in a unique identifying color. ‘Add Another Trace To Display’ Dialog Box Use the color-coded Active Trace buttons to choose the trace you want to be able Active Trace to manipulate in the Data Window.
Page 116 DISPLAYING DATA If you have overlaid a number of traces, the Scale to Active item becomes Scale to Active available on the Display Menu. Selecting Scale to Active means that all the data traces in your Data Window will be plotted against the scales of the active trace. Vertical axes will be rescaled even if the units do not match those of the active trace.
Page 117 DISPLAYING DATA bars, placed end to end. The upper scroll bar allows you to move between the members of the series, while the lower scroll bar allows you to view the traces that make up the particular member of the series. For further functionality associated with 3D Display Mode see also: Zooming In, Zooming Out and Scrolling and Reset under 2D previously;...
Page 118: Region Of Interest
REGION OF INTEREST Region Of Interest Region of Interest is an important post-acquisition tool, used for quantitative analysis of data. Region of Interest, abbreviated to ROI, can be selected from the Display menu on the Main Window, or by clicking the ROI icon the task bar in the Main Window.
Page 119 REGION OF INTEREST Region of Interest Buttons Along the bottom of the ROI Dialog Window are a series of buttons used for manipulating the ROI data. It will be useful to explain the functions of each these buttons. If you click on this button, the red ROI box will appear on your image. Use the Add ROI corner handles to drag out the size and location of your ROI.
Page 120 REGION OF INTEREST Clicking this button will present and group your ROI data, according to value Value View regions – mean, range, variance, and standard deviation. It will also display the pixel co-ordinates for all the ROI’s for that scan. Clicking the Live Update button will allow you to receive and calculate ROI data, Live Update while the system is running in Real Time Mode.
Page 121: Using The Command Line
USING THE COMMAND LINE Processing Data via the Command Line The Command Line Dialog Box is launched from the Command Line option on Command Line the Command Menu or from the Command Line Button on the Main Window. The Command Line allows you to enter one-line commands that are written in the programming language.
Page 122: Entering Hardware Details
ENTERING HARDWARE DETAILS Configuring the System for your Hardware The Hardware Menu allows you to configure the system so that the application Hardware software interfaces correctly with its hardware environment. The following options are provided: • Shutter Control launches a Shutter Control Dialog Box, which you can use to indicate when and how a hardware shutter should be used.
Page 123: Shutter Control
SHUTTER CONTROL The Shutter Control Dialog Box provides you with a selection of options that Shutter Control determine when and how a hardware shutter should be used. Launch it by selecting the Shutter Control option on the Hardware Menu. Certain settings (e.g. Permanently Open and Permanently Closed) take effect as soon as you close the Shutter Control Dialog Box by clicking the OK button.
Page 124 SHUTTER CONTROL General Shutter Control Radio buttons on the Shutter Control Dialog Box give you high-level control over the shutter: Fully Auto is the simplest shutter mode, as it leaves all shuttering decisions to the Fully Auto system. When you perform Take Signal, (see SECTION 4.4 DATA ACQUISITION) the shutter opens for the duration of the Exposure Time you have entered on the Setup Acquisition Dialog Box (see SECTION 4.5 SETTING THE ACQUISITION).
Page 125 SHUTTER CONTROL CCD has been supplied with a built in shutter, the TTL pulse is factory set to high. Changing this setting will affect operation of the shutter. • If you click the TTL Low button the system will cause the output voltage from the controller card to go ‘low’...
Page 126 SHUTTER CONTROL sequence, introducing an appropriate delay between the start of the shutter closed state and the commencement of the data being read out. If you do not have a shutter connected, set the Time to open or close to 0. Setting the Time to open or close to any other value will insert extra delays into cycle time calculations.
Page 127: Setting The Temperature
SETTING THE TEMPERATURE Launch the Temperature Control Dialog Box by selecting the Temperature option Temperature on the Hardware Menu, or by clicking the Thermometer icon at the bottom of the (Cooler Off) Main Window. The Temperature Control Dialog Box allows you to: (Cooler On) •...
Page 128: Cooling Performance And Operation
SETTING THE TEMPERATURE Cooling Performance and Operation: Technical Description The CCD is cooled using a thermoelectric (TE) cooler. TE coolers are small, electrically powered devices with no moving parts, making them reliable and convenient. A TE cooler is actually a heat pump – that is, it achieves a temperature difference by transferring heat from its ‘cold side’...
Page 129 SETTING THE TEMPERATURE Air Cooling Performance Air cooling is the most convenient method of cooling, but it will not achieve as low an operating temperature as water cooling (see below). Even with a fan, a heatsink typically needs to be 10°C hotter than the air (room) temperature to transfer heat efficiently to the surrounding air.
Page 130 SETTING THE TEMPERATURE Head Overheating Whichever cooling method you are using, make sure that the detector head does not overheat, as this can cause component failure. Overheating may occur if: The air vents on the sides of the head are accidentally blocked. You are using air cooling and have selected an operating temperature that is lower than the minimum achievable temperature.
Page 131 SETTING THE TEMPERATURE Water Cooling The table below is a guide to the minimum CCD operating temperatures for various water temperatures. Performance of individual systems will vary slightly. Evacuated Housing-High Performance Cooling with Power Supply Block (PSB) Water Temp External PSB -75°C -73°C -71°C...
Page 132: Organizing Your Windows
ORGANIZING YOUR WINDOWS The Window Menu offers a number of options which help you manage the Window windows or icons that you have created while working with the system. The following options are available: Cascade arranges any open Data Windows or Program Editor Windows (see Cascade SECTION 6 WORKING WITH PROGRAMS) into a stack which runs diagonally across the Main Window and which allows you to see the title bars of each of the windows...
Page 133: Copy To Clipboard
ORGANIZING YOUR WINDOWS (For use only with Data Windows.) Copy to Clipboard copies the contents of the Copy to Clipboard active Data Window into the Windows ‘clipboard’ - the store in which Windows normally holds data that has been cut or copied. Data, which have been copied to the clipboard, can be pasted into other Windows applications, where it will be treated as a picture or ‘bitmap’.
Page 134: Using The Remote Control
USING THE REMOTE CONTROL The Remote Control allows various commands of the iXon software to be modified using an infra-red signal sent to a Receiver. The range of operation of the remote control is up to a distance of 12m from the receiver along the line of sight. Connecting the Remote •...
Page 135 USING THE REMOTE CONTROL Using The Remote The remote control has two modes of operation. The first provides direct control over Control the Run Time Control dialog box (see next page), where the following parameters may be varied: • • Run/Abort acquisition Exposure Time EM Gain...
Page 136 USING THE REMOTE CONTROL Operating The Run Time To open the Run Time Control dialog box, press the Run Time Control button on Control Dialog Box the toolbar of the main software window. The default settings of the dialog box are shown below.
Page 137 USING THE REMOTE CONTROL Button B (the unmarked black button on the main panel of the remote) switches between the two modes of operation of the Remote Control. Press the button to either control the Run Time Control dialog box exclusively or to operate the windows pointer.
Page 138 WORKING WITH PROGRAMS Introduction 5 . 1 Editing Programs 5 . 2 Running Programs 5 . 3 Section 5...
Page 139: Andor Basic
WORKING WITH PROGRAMS … Working with Programs Andor Basic programming language allows you to create programs for Andor Basic customized control of data acquisition and customized manipulation of data. See the Programmer’s Guide to Andor Basic The system provides facilities to let you Edit, Save and Run your programs. The Program Editor Window Opening a program file, or selecting New Program from the File Menu (see SECTION 2 INSTALLATION) launches a Program Editor Window where you can...
Page 140: Editing Programs
EDITING PROGRAMS Edit Cut or Copy text that you have highlighted. Paste the text into a new position. Copy Paste inserts cut or copied text into the position following the cursor, or replaces Paste text that you have highlighted. Undo causes the text to revert to its state before the last change was made. Undo Search •...
Page 141 EDITING PROGRAMS • In the text box marked Find what, type the search string. In the text box Replace marked Replace with, type the word or phrase that you want to use instead. • Click the Find Next button to highlight the next occurrence of the search string. Find Next •...
Page 142: Running Programs
RUNNING PROGRAMS To run a program, make sure you have opened the appropriate .pgm file and ensure Run Program that the file name appears in the drop-down list box next the Run Program button; then click the Run Program button. To change the name of the file you want to run: either open the drop-down list and click the name of the file;...
Page 143: Third Party Sofware
THIRD PARTY SOFTWARE Section 6...
Page 144 THIRD PARTY SOFTWARE Soft Imaging System 6 . 1 Other S/W Supported 6 . 1 Section 6...
Page 145: Soft Imaging System
THIRD PARTY SOFTWARE Soft Imaging System Andor's preferred imaging partner is SiS, Soft Imaging System, the imaging software specialists. We are working together to provide you with imaging software for all your imaging requirements. AnalySIS software, a flexible software package has been designed to work in conjunction with your Andor camera with minimum effort.
Page 146: Tutorials
TUTORIALS Tutorial 1: Measuring Gain Factor at a Given Temperature Setting Using a Weak 7 . 1 LED Light Source Tutorial 2: Extending Dynamic Range in ‘Slow Scan’ Measurements By Accumulating 7 . 2 Signal Tutorial 3: Performing Basic Low-Light Microscopy Functions Using the iXon 7 .
Page 147: Tutorial 1
TUTORIAL 1 … Measuring Gain Factor at a Given Temperature Setting Using a Weak LED Light Source Additional Requirements Stable LED light source, Compatible camera lens, Darkened ambient conditions, Optional - Neutral density filters The object of this experiment is to make an approximate measurement of the true gain factor applied at a series of working empirical gain settings.
Page 148 TUTORIAL 1 Alternatively, neutral density filters may be inserted in front of the light source, or the aperture on the lens may be lessened in order to reduce the intensity of light reaching the sensor. If the signal appears too low, relative to the surrounding noise floor, the exposure time may be increased accordingly.
Page 149: Tutorial 1
TUTORIAL 1 Acquisition menu select Take Background. Background data is displayed on-screen under the Bg tab of an Acquisition Window. Turn the light source on again. Now acquire the signal image for this gain setting – from the Acquisition menu on the Main Window, select Take Signal (or alternatively click the Take Signal button).
Page 150 TUTORIAL 1 can be plotted and extrapolated, from which gain factors at any gain settings can readily be estimated at this system temperature. Section 7. 1 Page 150...
Page 151 TUTORIAL 2 Extending Dynamic Range in ‘Slow Scan’ Measurements By Accumulating Signal Although the iXon camera, due to it’s high-speed operation and ultimate Background EMCCD sensitivity performance, is very well suited to dynamic low light measurements, it can just as readily be applied to ‘slow scan’ non-dynamic quantitative measurements such as microtitre plate, gel or biochip reading.
Page 152 TUTORIAL 2 determine approximately how close the most intense pixels are to the saturation level. If this signal level is less than the saturation limit of 16384 counts, the exposure time may be proportioned up accordingly to approach this maximum value. If the signal is already saturating the CCD, the exposure time will have to be reduced and the above procedure repeated.
Page 153 TUTORIAL 2 (for sensors without anti-blooming, the weakest signal should be sufficiently spaced within the image from the stronger signals, such that charge does not ‘spill over’ into a measured region). Measure the S:B ratio of the weakest signal using ROI, as in the above step, and repeat this procedure until an optimal exposure time (E2) has been determined which meets the S:B requirements of the assay.
Page 154 TUTORIAL 2 Andor software – from the Command menu on the Main Window, select the option Command Line. Type in the command - ?gettemperature() – and click on Execute. Furthermore, with electron multiplying technology, operation at a lower temperature reduces darkcurrent, which would otherwise be amplified in the gain register and appear as spurious noise spikes (the extent of which depends on exposure time).
Page 155 TUTORIAL 3 Performing Basic Low-Light Microscopy Functions Using the iXon Additional Requirements Epifluorescence Microscope and camera adaptor. Optical filter set matched to fluorescence beads. Fluorescent microspheres, e.g. Fluorescein containing micron scale polystyrene FluoSpheres® from Molecular Probes. Darkened ambient conditions. The object of this experiment is to perform some basic low light microscopy functions with the iXon, including recording kinetic series and determination of optimal gain, utilising as a simple example, micron scale fluorescent microspheres in solution.
Page 156 TUTORIAL 3 Record a Kinetic Series - From the Acquisition menu on the Main Window, select Setup Acquisition. In Acquisition Mode select Kinetic Series, and type in the number of frames to be recorded on the run, e.g. 50. In the Setup CCD tab, ensure gain is set to zero and set exposure time to approximately 10-20 ms (the Kinetic Cycle Time will update as exposure time is changed, as will the indicated Frame Rate in Hz).
Page 157 TUTORIAL 3 The row of mean values can be highlighted for any ROI then plotted vs. either frame number or time by clicking the appropriate icon in the ROI display window. The kinetic plot will appear in a new window. In the present example, the fluctuations in the plot over time represent fluorescent beads passing through or coming into focus within the field of view.
Page 158 TUTORIAL 3 increased (this may be more easily visualised by clicking the cursor in a background region of the image, then selecting the 2D icon), it is likely that background light is being leaked into the system. This can markedly reduce the effectiveness of this technology, since leaked background or unwanted background fluorescence is primarily contributing to the detection limit at higher gain settings, rather than the camera readout noise.
Page 159 TUTORIAL 3 (such as was recorded in video mode) in order to determine the position of the sides of a sub area (square or rectangle) encompassing the fluorescent beads of interest. In the Setup Image tab of the Setup Acquisition Dialog box, select Custom Sub-Image and enter the sub-area dimensions into the left, right, top and bottom boxes.
Page 160: Appendices
APPENDICES...
Page 161 APPENDICES The Controller Card 8 . 1 Windows Glossary 8 . 2 Imaging Glossary 8 . 3 Terms and Conditions 8 . 4 Section 8..
Page 162: The Controller Card
THE CONTROLLER CARD Controller Card: A uxiliary Connector Description PinNum bers PinDescriptions 5 4 3 2 1 Aux in 1 I2C Data Aux in 3 Aux in 2 Aux in 4 10 9 8 7 6 15 14 13 12 11 I2C Clock G N D Aux out 1...
Page 163 THE CONTROLLER CARD · AUX OUT – Pins 6, 7, 8 and 14 provide up to 4 auxiliary output channels. The outputs are SN74128 50 Ù TTL output driver that can be controlled via the Andor Basic language. WARNING: Pins 11, 12 and 13 are RESERVED, these pins are not available for auxiliary use.
Page 164: Windows Glossary
WINDOWS GLOSSARY As far as possible the User’s Guide employs standard Windows terminology to describe the features of the user interface. If you are unfamiliar with Windows, the definitions that follow cover some of the most significant terms used in this User’s Guide.
Page 165 WINDOWS GLOSSARY The window which you are currently working with is known as the Active Active Window Window. It is always positioned in the foreground of your screen. By the bolder color of its title bar, the active window is distinguished from other windows you may have open.
Page 166: Menu Bar
WINDOWS GLOSSARY A Menu is a list of commands that you can select by clicking. The menu titles are Menu; Menu Bar arranged in a row, or Menu Bar, across the top of a window. Clicking the menu title causes the menu itself to ‘drop down’, allowing you to select the option you require.
Page 167: Imaging Glossary
IMAGING GLOSSARY Accumulation is the process by which data that have been acquired from a number Accumulation of similar scans are added together in computer memory. This results in improved signal to noise ratio. An Acquisition is taken to be the complete data capture process that is executed Acquisition whenever you click Take Signal, Take Background, or Take Reference on the Acquisition Menu or whenever you click the Take Signal button (see...
Page 168 IMAGING GLOSSARY The system uses the two main binning variants, vertical and horizontal binning. Binning Patterns Imaging: The default imaging mode is Full Resolution Image, which causes a value to be read out for each pixel on the CCD-chip. However, the Image Dialog Box, accessible from the Setup Acquisition Dialog Box, lets you define superpixels which in effect represent the charge binned (by means of a combination of vertical and horizontal binning) from groups of pixels (each superpixel being a rectangle of a...
Page 169: Exposure Time
IMAGING GLOSSARY generally remove most of the shot noise caused by dark signal. The Detection Limit is a measure of the smallest signal that can be detected in a Detection Limit single readout. The smallest signal is defined as the signal whose level is equal to the noise accompanying that signal, i.e.
Page 170 IMAGING GLOSSARY signal; and shot noise from the light signal itself. Shot noise cannot be removed because it is due to basic physical laws. Most simply defined, shot noise is the square root of the signal (or dark signal) measured in electrons. Readout noise (which in our detectors is, in any case, low) is due to the amplifier and electronics: it is independent of dark signal and signal levels;...
Page 171 IMAGING GLOSSARY The glossary refers to signals as a number of electrons. More strictly speaking these Quantum Efficiency; are ‘photoelectrons’, created when a photon is absorbed. When a UV or visible photon is absorbed by the detector it can at best produce only one photoelectron. Spectral Photons of different wavelengths have different probabilities of producing a Response...
Page 172: Shift Register
IMAGING GLOSSARY The Shift Register usually consists of a single row of elements (or pixels) running Shift Register parallel to and below the bottom row of light-gathering pixels (the image area) on the EMCCD-chip. The shift register is protected from light by an aluminum mask. The elements in the shift register have a greater capacity to store charge (a greater ‘well depth’) than the other pixels on the EMCCD-chip.
Page 173: Terms And Conditions
‘GOODS’ means the goods (including any instalment of the goods or any parts for them), which the Seller is to supply in accordance with these Conditions. ‘SELLER’ means Andor Technology Limited. ‘CONDITIONS’ means the standard terms and conditions of sale set out in this document and (unless the context otherwise requires) includes any special terms and conditions agreed in writing between the Buyer and Seller.
Page 174: Index
INDEX Section 9..
Page 175 INDEX Dark Signal, 168 Background Correction, 90 A/D conversion, 167 Fixed Pattern Noise, 170 Absorbance Units, 91 reduced by Cooling, 127 Absorptance, 90 Shot Noise, 173 Absorption Coefficient, 91 Darksignal, 47 Accumulated Kinetic Series, 71 Data * Reference, 92 Accumulation, 70 Data Range Active Trace, 115 and Brightness, 103...
Page 176 INDEX Replace Program Editor Window, 141 Maintain Aspect Ratio, 106 Reset, 113 Run Program Andor Basic, 142 Noise , 46 Saturation, 172 Scan, 172 Setting the Temperature, 127 Overheating Setup Acquisition, 67 Causes, 130 Setup Data Type, 64 Overlay, 114 Shift Register, 173 Overlay and Keep, 115 Shortcuts, 29...

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