Andor iQ Software

Functionality

• Powerful Wizards Help with Settings and Configuration
• Powerful Processing
• High-Performance Off-Line Processing
• Targeted Analysis Tools and FlexiblePlug-in Architecture
• Multi-dimensional Views and Imaging Options

Powerful Wizards Help with Settings and Configuration

Protocol tree shows acquisition sequence control and enables easy editing of parameters

• Channels Wizard lets you manage illumination and imaging for any illuminescent, fluorescent or brightfield imaging configuration.
• Scan Wizard creates XY and Z Scans.
• Protocols define control acquisition sequence, timing, triggering and event monitoring.
• Rapid Setting and Protocol editing via right mouse click pop-up menus.
• Create and manage experiment acquisition protocols for easy re-use and sharing.
• Z-offset between channels to compensate for optical chromatic errors.
• Control and synchronization to external hardware via programmable Trigger and Event interface (PCU).
• Integrate with Electrophysiology Perfusion, Flash Photolysis and FRAP Solutions.

iXon^EM+ DU897-BV and iQ software is used to image calcium with Fura2. Dr Fayuk studies neuron receptors in brain sections and cultures to elucidate processes of addiction. In this example a cultured neuron was patch-clamped and an iontophoretic electrode used to generate a 50ms Choline injection (200nA) at the spot shown. Synchronization to electrophysiology apparatus is achieved with Andor’s iQ software and Precision Controller Box (PCB) Courtesy Drs Dmitriy Fayuk and Jerry Yakel"s Neurobiology Laboratory, NIEHS, NC.

4D maximum intensity image with pseudocolor shows live neuron labelled with quantum dots

Powerful Processing

On-Line Analysis and Charting

• On-line Data is continuously streamed to a disk file. Charting of multi-channel, multiregion, multi-field or multi-well studies is possible, with optional ratio mode.
• Histogram analysis of multiple user-defined regions of interest.
• Line profiling for single or multi-channel image data on Live or stored data.

High-Performance Off-Line Processing

• Ratio analysis for dual channel analysis, time-series pairing and self ratio (with standing level from first n frames). Background correction and Mask image can be used to reduce noise in low signal regions. Ion concentration calibration options using either Grynkiewicz or Almers-Neher methods for ratio dyes.
• Kymograph tool allows oblique line and polyline probing of multidimensional data, with preview. Now with "Lamella" mode for protrusion/contraction rate analysis.
• Dual field splitting and alignment tool to allow analysis of simultaneous captured two or four channel data. Now with live field subtraction alignment tool.
• Multi-dimensional Math: difference, average, encode and projection processing.
• Ergodic averaging: improve SNR from repetitive signal acquisition and processing.
• Photobleaching compensation: minimizes effect on analysis results or visualization for AVI (movie) creation.

Targeted Analysis Tools and Flexible Plug-in Architecture

• Plug-in architecture allows for in-field updating and access to new processing modules as they become available. This structure replaces our legacy product, Lucida, as all previous functionality is migrated to the newer iQ software architecture. We now offer iQ-WS to function on satellite workstations. We are already planning a future with scripting to allow automation of many processing and analysis tasks.
• Andor's ClearView Deconvolution product is supplied as a plug-in option and provides powerful constrained iterative algorithms for image resolution enhancement, haze removal and preparation of wide field microscopic images for 3D/4D visualization.
• Region of Interest (ROI) feature definitions enhance ROI data manipulation and interpretation.
• Thru-series analysis: region-based, time, Z or spectral dimensions. Analyze single or multi-channel image data with optional background correction.
• Text and Dimension stamp overlays for monitoring and export of data.
• Extend analysis dynamic range: "greys per second" mode.
• Co-localisation analysis of multi-channel, RGB image data.
• Tracking: automatic and interactive tools for monitoring motion.

Multi-dimensional views and imaging options

Multi-Channel Views, Voxel Rendering, Orthogonal Viewing and support for confocal instruments

Andor iQ offers powerful visualization tools as standard, with 3D, 4D and 5D capabilities. Raw data from 3D, 4D, 5D and 6D acquisition protocols, can generate image data fast and iQ provides powerful tools for rapid data visualization and analysis.

• Navigator - Explore the data with a multidimensional movie player interface and lets you view multi-channel data in overlay mode, merging imaging modes required.
• MC View - View and manipulate multi-channel data in an overlay mode. Channel contrast and balance are individually user-controlled.

3D/4D Viewer

Single section and voxel rendering of a 3-channel z-series, acquired on a Bio-Rad confocal instrument, viewed in multi-channel mode in iQ software

Voxel renderer methods: maximum intensity, minimum intensity, opacity blending and first pixel on ray trace. Options include:

• Intensity range
• Multi-channel with scaling/mapping to enhance features and study spatial coincidence.
• Wire-frame dragging adjusts angle of view.
• Animation method interpolates between user-defined key frames (and Zoom and Time points for 4D data) to create a movie.
• Export to AVI files through the movie editor.

OrthoView and Kymopgraph

Create orthogonal and oblique slices through multi-dimensional images and visualize as images. Kymograph allows Tracking of waves and features and new Lamella mode enables extension/contraction rate analysis of e.g. filopodia and ruffling.

Strip View

Cultured neurons were loaded with DiSBAC2(3), a voltage sensitive dye and visualized with the Revolution 488. Exposure time of the iXon 887-BV was 100ms and EM Gain 200. The figure shows a film strip from Andor iQ of maximum intensity projection time series. Time point 4 is taken almost simultaneously with the addition of KCl, which depolarizes the cells and shows a near instantaneous rise in DiSBAC2(3) signal. ΔF of almost 45% is observed in this example. Imaged at The 3D Microscopy of Living Cells Course, Vancouver 2005. Courtesy Dr Iain Johnson, Invitrogen, Eugene, OR.

Film strip view of multidimensional images. Control frame size, number of columns, selected dimension, range and increment between frames, border color and width. Images can be shown with dimension stamps e.g. time of acquisition or Z-depth.

Montage of blood vessel section created from 108 (12x9) fields, acquired with iQ-MF controlling iXon DV885 EMCCD camera. Montage scanning included in iQ-MF module

Montage Imaging

iQ-MF module includes a Montage scan mode. Strip View lets you visualize the montage of large specimens acquired at high power, as shown in the figures.

iQ-MF imaging also allows time lapse recoding from multiple fields of view, so that data can be gathered on a large number of cells, or multiple treatment groups, increasing 'n' for statistical analyses or speeding experimental throughput or both.

Movie Editor

Courtesy Dr Lance Prince’s lab in Pediatrics, UA Birmingham, AL. Dr Prince is studying lung development in live explants to better understand foetal lung development and pathology. Andor iQ hosts his live cell imaging system.

A multi-dimensional filehandling interface for Windows AVI files. Choose start-end frames and animation dimension to create focus, spectral or time-series movies. Movie Editor provides ROI,overlay and multi-channel options to simplify production of presentation data.

File-import-export capabilities

AVI: images from iQ can be exported via Windows CODECS for Powerpoint etc. Andor iQ will read AVI files.

BioRad: file format support for confocal and multi-photon microscope systems.

ICS: International Cytometry Standard files - multi-dimensional images.

TIFF: most single frame and multi-frame formats supported.

Zeiss LSM: multidimensional file support for the Zeiss LSM 510 confocal instruments.