Voltage Sensitive Dye Based Imaging System

  • Overview

  • Uses and Technical Overview

  • Common Features of All Models


Software tools

  • VDAQ 2.4- Data Acquisition Software

  • Data Analysis Software



The OI Imager 3001 image data acquisition and analysis system carries out optical imaging based on both intrinsic optical signal and voltage sensitive dye (VSD) signals-as well as on signals from calcium dyes and other extrinsic optical probes. The Imager 3001 can monitor blood volume and flow changes, as well as arterial pulsation and the respiratory motion of cortex. Explorations of cortical microcirculation with the Imager 3001 may also help improve the spatial resolution and interpretation of functional MRI and PET imaging on human subjects, or even lead to MRI imaging based on activity-dependent changes in new physiological parameters.
The Imager 3001 is not a bare-bone camera-and-frame grabber combination, leaving the user to figure out the means of adapting it to experimental use. Rather, OI offers and supports it as an integrated solution specifically designed for intrinsic optical signal, VSD, and other types of neuronal optical imaging. OI also offers many imaging accessories for use with the Imager 3001, and an on-site installation program to help you get your imaging experiments up and running.
Not only do the imaging functions of the Imager 3001 complement each other, they can also complement non-imaging techniques that may already be used in your lab. Imaged maps provide information about functional context that can be used to guide site selection for electrode recordings (intracellular or extracellular), or to target micro-stimulation, tracer injections, or other experimental manipulations. Designed from the experience of working scientists, the Imager 3001's laboratory interface can communicate with much of the standard equipment found in neurophysiology and biophysics laboratories.

Uses and Technical Overview

Intrinsic Optical Signals
As an intrinsic signal imager, the Imager 3001 system detects tiny intrinsic changes in the optical properties of electrically and/or metabolically active brain tissue (signals as small as 1 part in 10,000 are detectable with signal averaging). The pioneering work of Hill, Keynes, Chance, Jobsis and their colleagues first demonstrated the existence of these intrinsic signals nearly fifty years ago. However, since these signals are very small, their use for imaging of the functional architecture of cortex began only in 1986, as technology and techniques were improved. One source for the activity-dependent optical signal is a small change in the color of the tissue produced by changes in oxygen delivery from oxy-hemoglobin within the capillaries in response to metabolic demand. Other intrinsic signals originate from activity-dependent light scattering changes, and changes in the oxidation states of intrinsic chromophores such as cytochromes.
Intrinsic imaging has facilitated high-resolution imaging of the adult functional architecture of the cerebral cortex in the living brain of mice, rats, guinea pigs, gerbils, ferrets, cats, monkeys and humans. In some animals, activity maps have also been obtained through intact dura and thinned bone, which allows visualization of the development of the functional architecture of the cortex over long periods of time. Success has also been achieved with chronic recording paradigms in larger and adult mammals. Activity-dependent intrinsic signals have also facilitated in-vitro studies in brain slices, and in the isolated but intact mammalian brain.

Voltage Sensitive Dye
The term "voltage sensitive dye" (VSD) is used for compounds that act as optical transducers of membrane potential changes. These probes are used to stain a living preparation. Applied to the brain, they bind to the external surface of the membranes of living cells without interrupting their normal function. Once introduced into a preparation, VSDs rapidly (within a microsecond) alter the intensity and/or wavelengths of fluorescent light they emit as a function of changes in neuronal membrane potential.
Recent improvements in the VSD probes available for in vivo VSD imaging (see the Voltage Sensitive Dyes product sheet for more information) have dramatically increased the signal-to noise-level obtainable-by as much as 10-30 times over previous probes. Combined with the Imager 3001, in vivo or in vitro, voltage sensitive dyes can now provide both fast time resolution and the spatial resolution required to visualize rapid, complex spatio-temporal patterns of neuronal activity.

Calcium Dyes and Other Optical Probes
Calcium signals are usually more than an order of magnitude larger than the VSD signal. Due to this fact, the Imager 3001/M New back thinned 100% fill factor high-resolution camera performs well in detecting small calcium signals, although it is optimized for high light level applications.
See OI's technical white papers for more information on all these imaging techniques

Imager 3001: A Multi-Purpose Approach

The Imager 3001's multi-purpose design is a powerful approach to optical imaging. The intrinsic optical signal and VSD imaging techniques each excel at revealing different information about the brain's functional architecture. Intrinsic optical signal imaging is particularly suited to revealing high-resolution spatial features of the brain's functional architecture. VSD imaging excels at revealing the temporal structure of neuronal responses across regions of exposed brain. And calcium imaging extends the power of the system further. Irrespective of the signal used to extract physiological information, the Imager 3001 is well suited to study nearly all the preparations used in brain research explorations.

Common Features of All Models

All Imager 3001 models include the following:

  • VDAQ 2.4
    The latest version of our time-tested data acquisition system running under Windows XP?. It allows for a wide variety of experimental paradigms, live image display for focusing, live display of enhanced differential images, feedback during experiments with flexible on-line analysis displays, and an exceptionally intuitive user interface.
  • WinMix 2.0
    The latest version of our flexible data analysis software, and easy enough to use that it can provide critical information while the experiment is still in progress. WinMix includes utilities to convert or view VDAQ data files or movies, a scripting component, easy-to-use interactive image processing tools, and various export capabilities that let you transport your data or images to 3rd party software.
  • Imaging Camera
    Camera type varies by model, details are provided in our product sheets.
  • Laboratory Interface Unit
    All Imager 3001 systems include a laboratory interface panel that allows you to connect the imager to standard experimental apparatuses such as stimulators, shutters, respirators, etc. All systems also include an "artificial cortex" target for calibration and testing. The BNC Panel allows you to connect your data acquisition system to standard laboratory equipment.
  • An optional Analysis Workstation computer, so that you can perform fast WinMix data analysis while continuing to collect data.
  • All systems are delivered in rugged rack-mount chassis with heavy-duty power supplies and late model motherboards, generous amounts of RAM and disk space, high-speed CPUs, and roomy RAID0 disk arrays for fast data access.
  • All models are turn-key systems. They arrive with all the necessary cables and software, with all software installed and tested. Installation of the cables is shown in an illustrated Cable Installation Guide, so you can be up and running in a very short time.
  • Systems containing both an Acquisition Workstation and an Analysis Workstation include a network hub to interconnect the two computers.
  • ADAQ Option
    ADAQ, an integrated Analog Data AcQuisition facility for VDAQ, lets you make electrical recordings that are co-triggered with your image recording. You can collect up to 16 channels of electrical activity (or 8 differential channels) at speeds of up to 500,000 samples/second, with 12-bit resolution. ADAQ data files are similar in format to existing VDAQ block data files, and we will offer enhanced versions of our analysis tools that can work with analog waveform data.

    See our Accessories product sheets for imaging items sold separately.
    Monitors included by special order only.

To learn more about current Imager 3001 models, see product sheets & product pages in PDF formats below:

Imager 3001/M

IMAGER 3001/M Product Sheet

Imager 3001/S
IMAGER 3001/S Product Sheet

Imager 3001/C
IMAGER 3001/C Product Sheet

Imager 3001/Celox
IMAGER 3001/Celox Product Sheet


VDAQ 2.4- Data Acquisition Software

VDAQ 2.4 (Video Data AcQuisition) is the next-generation data acquisition component of the Imager 3001, running under Windows XP?. VDAQ runs intrinsic and voltage sensitive dye imaging experiments as well as calcium dye and other extrinsic probe experiments. It controls image acquisition, as well as external devices such as a stimulator, respirator, and illumination shutter. All experimental parameters can be saved to disk, to allow easily restoring experimental settings from session to session.

Other external hardware devices can also be controlled and monitored as required according to the experiment. Alternatively, VDAQ can itself be controlled through a serial port connection for awake behaving monkey experiments. VDAQ also has functions for performing the setup tasks that are performed at the beginning of an experimental imaging session. It provides on-line image analysis functions that are critical to monitoring and evaluating the progress and quality of experiments as they unfold. Finally, VDAQ contains the functions used to calibrate and test the video, optical, and image processing components of the instrument.

LongDaq software for continuous imaging

LongDaq collects brain imaging data continuously, limited only by the speed and free space of the available storage. It's available as either an add-on to a Vdaq system, or as a system in its own right.
LongDaq, Optical Imaging's cost-effective continuous imaging software, now supports a stim-map feature that provides fine-grained control of stimulus output bits during a continuous imaging session. This provides useful synchronization information and lets LongDaq trigger stimulators and other devices during acquisition.
LongDaq's useful Quality Assurance data is now saved automatically each time data is collected.
Both Vdaq and LongDaq now allow for stimulus input information to be provided via UDP network packets. Previously, such information could only be provided using TTL level inputs.


Data Analysis Software

WinMix is a data analysis suite specifically designed to handle data from optical imaging experiments, including those generated using VDAQ. It consists of the new Block Mix scripted data analysis program, plus a combination of powerful interactive tools including Block View and Block Convert. The powerful Block Mix script tool includes display capabilities during analysis, logging of statistics of output images, and syntax highlighting in the edit window.

WinMix's interactive tools include:

  • Viewers for image and data files.
  • An image-processing component for filtering, image histograms, line profiles, image arithmetic
  • A polar analysis module for producing polar plots from groups of image files
  • A MovieLoop facility for comparing pairs of images, or working with movies of data
  • Image conversion to a variety of popular image formats, including BMP, TIF, and JPEG

Block View
Block View is an interactive program for viewing the data files produced during an imaging experiment. Its various display options and simple processing options let you use it for preliminary data analysis. Block View also lets you display a movie of specified frames, and it can compute superpixel graphs showing the average value of a region over the course of many frames.

Block Convert
Block Convert is a powerful data conversion and compression program. Block Convert performs spatial and temporal binning, to improve the efficiency of subsequent analysis operations. Other conversions include accumulation of block files, selection of specific conditions, and cropping the data to a smaller region of interest. The output format can be either an OI block file (which can be viewed with Block View, or further processed with WinMix), a series of floating point image frames stored in a standard, compressed ZIP archive, or a MatLab? format data file.



Featured Publications on OI Brain Imagers

  • Palagina G, Eysel UT and Jancke D (2009) Strengthening of lateral activation in adult rat visual cortex after retinal lesions captured with voltage-sensitive dye imaging in vivo, PNAS 106 (21):1743-1747.
  • MacEvoy, SP, Tucker TR and Fitzpatrick D (2009) A precise form of divisive suppression supports population coding in the primary visual cortex, Nature Neuroscience 12 (5): 637-645.
  • Livneh Y, Feinstein N, Klein M and Mizrahi A (2009) Sensory Input Enhances Synaptogenesis of Adult-Born Neurons, Journal of Neuroscience 29 (1): 86? 97.
  • Li Y, Van Hooser SD, Mazurek M, White LE and Fitzpatrick D (2008) Experience with moving visual stimuli drives the early development of cortical direction selectivity, Nature 456 (7224): 952-956.
  • Chen G, Lu HD and Roe AW (2008) A Map for Horizontal Disparity in Monkey V2, Neuron 58 (3):442-450.
  • Nauhaus I., Benucci A, Carandini M and Ringach DL (2008) Neuronal Selectivity and Local Map Structure in Visual Cortex, Neuron 57 (5): 673-679.
  • Kerr JND,de Kock CPJ, Greenberg DS, Bruno RM, Sakmann B. and Helmchen F (2007) Spatial Organization of Neuronal Population Responses in Layer 2/3 of Rat Barrel Cortex, Journal of Neuroscience 27 (48): 13316-13328.
  • Farley, BJ, Yu H, Jin DZ and Sur M (2007) Alteration of Visual Input Results in a Coordinated Reorganization of Multiple Visual Cortex Maps, Journal of Neuroscience 27 (38): 10299-10310.
  • Chen LM, Turner GH, Friedman RM, Zhang N, Gore JC, Roe AW and Avison MJ (2007) High-Resolution Maps of Real and Illusory Tactile Activation in Primary Somatosensory Cortex in Individual Monkeys with Functional Magnetic Resonance Imaging and Optical Imaging, Journal of Neuroscience 27 (34): 9181-9191.
  • Accolla R, Bathellier B, Petersen CCH and Carleton Alan (2007) Differential Spatial Representation of Taste Modalities in the Rat Gustatory Cortex, Journal of Neuroscience 27 (6): 1396-1404.
  • Xu X, Collins CE, Khaytin I, Kaas JH and Casagrande VA (2006) Unequal representation of cardinal vs. oblique orientations in the middle temporal visual area, PNAS 103 (46): 17490-17495.
  • Chen Y, Geisle WS and Seidemann E (2006) Optimal decoding of correlated neural population responses in the primate visual cortex, Nature Neuroscience 9 (11): 1412?1420.
  • Ohki K, Chung S, Kara, P, Hubener M, Bonhoeffer T and Reid RC (2006) Highly ordered arrangement of single neurons in orientation pinwheels, Nature 442 (7105): 925-928.
  • Lin DY, Shea SD, Katz LC (2006) Representation of Natural Stimuli in the Rodent Main Olfactory Bulb, Neuron 50 (6): 937-949.
  • Hofer SB, Mrsic-Flogel TD, Bonhoeffer T and Hübener M (2005) Prior experience enhances plasticity in adult visual cortex, Nature Neuroscience 9 (1): 127-132.
  • Roe AW, Lu HD and Hung CP (2005) Cortical processing of a brightness illusion, PNAS 2005 102 (10): 3869-3874.
  • Van Hooser SD, Heimel, JAF, Chung S, Nelson SB and Toth LJ (2005) Orientation Selectivity without Orientation Maps in Visual Cortex of a Highly Visual Mammal, Journal of Neuroscience 25 (1): 19-28.
  • Vanzetta I, Slovin H, Omer DB, et al. Columnar resolution of blood volume and oximetry functional maps in the behaving monkey: Implications for fMRI Neuron42 (5): 843-854 jun 10 2004
  • Eysel UT. Illusions and perceived images in the primate brain Science 302 (5646): 789-791 oct 31 2003
  • Petersen CCH, Grinvald A, Sakmann B. Spatiotemporal dynamics of sensory responses in layer 2/3 of rat barrel cortex measured in vivo by voltage-sensitive dye imaging combined with whole-cell voltage recordings and neuron reconstructions. Journal Of Neuroscience 23 (4): 1298-1309 feb 15 2003
  • Tucker TR, Katz LC. Spatiotemporal patterns of excitation and inhibition evoked by the horizontal network in layer 2/3 of ferret visual cortex. JOURNAL OF NEUROPHYSIOLOGY 89 (1): 488-500 JAN 2003
  • Bosking WH, Crowley JC, Fitzpatrick D (2002). Spatial coding of position and orientation in primary visual cortex. Nat Neuroscience, 5:874-882.
  • Luo M, Katz LC (2001). Response correlation maps of neurons in the mammalian olfactory bulb. Neuron, 32: 1165-1179.
  • Schwartz TH, Bonhoeffer T (2001). In vivo optical mapping of epileptic foci and surround inhibition in ferret cerebral cortex. Nature Med, 7:1063-1067.
  • Dragoi V, Rivadulla C, Sur M (2001). Foci of orientation plasticity in visual cortex. Nature, 411: 80-6.
  • Rubin BD, Katz LC (2001). Spatial coding of enantiomers in the rat olfactory bulb. Nature Neurosci, 4: 355-6.
  • Uchida N, Takahashi YK, Tanifuji M, Mori K (2000). Odor maps in the mammalian olfactory bulb: domain organization and odorant structural features. Nature Neurosci, 3:1035-43.
  • von Melchner L, Pallas SL, Sur M (2000). Visual behaviour mediated by retinal projections directed to the auditory pathway. Nature, 404:871-6.
  • Grinvald A, Slovin H, Vanzetta I (2000). Non-invasive visualization of cortical columns by fMRI (2000). Nature Neurosci, 3:105-7.
  • Crowley JC, Katz LC. Early development of ocular dominance columns (2000). Science. 2000 290(5495):1321-4.
  • Sharma J, Angelucci A, Sur M (2000). Induction of visual orientation modules in auditory cortex (2000). Nature, 404: 841-7.
  • Das A, Gilbert CD (1999). Topography of contextual modulations mediated by short-range interactions in primary visual cortex. Nature, 399: 655-61.
  • Vanzetta I, Grinvald A (1999). Increased cortical oxidative metabolism due to sensory stimulation: implications for functional brain imaging. Science, 286: 1555-8.
  • Das A, and Gilbert CD (1997). Distortions of visuotopic map match orientation singularities in primary visual cortex. Nature, 387:594-598.
  • Crair MC, Gillespie DC, Stryker MP (1998). The role of visual experience in the development of columns in cat visual cortex. Science, 279:566-70.
  • Weliky, M. and Katz LC (1997). Disruption of orientation tuning in visual cortex by artificially correlated neuronal activity Nature, 386:680-685.
  • Maldonado PE, Godecke I, Gray CM, Bonhoeffer T (1997). Orientation selectivity in pinwheel centers in cat striate cortex. Science, 276:1551-5.
  • Shoham, D, Hubener, M, Grinvald A and Bonhoeffer T (1997). Spatio-temporal
    frequency domains and their relationship to cytochrome oxidase staining in cat visual cortex. Nature, 385: 529-534.
  • Godecke I, Bonhoeffer T (1996). Development of Identical Orientation Maps for 2 Eyes without Common Visual Experience, Nature, 379: 251-254.
  • Weliky M, Bosking W, Fitzpatrick D (1996). A systematic map of direction preference in primary visual cortex. Nature 379: 725-728.
  • Malonek D, and Grinvald A (1996). Interactions between electrical cortical activity and the microcirculation revealed by imaging spectroscopy: Implications for functional brain mapping. Science 272: 551-554.
  • Wang G, Tanaka K and Tanifuji M (1996). Optical imaging of functional
    organization in the monkey inferotemporal cortex. Science, 272: 1665-1668.
  • Sheth BR, Sharma J, Rao SC, and Sur M (1996). Orientation maps of subjective contours in visual cortex. Science 274: 2110-2115.
  • Das, A Gilbert, CD (1995). Long-range horizontal connections and their role in cortical reorganization revealed by optical recording of cat primary visual cortex. Nature, 375: 780.


  • White LE and Fitzpatrick D (2007) Vision and Cortical Map Development, Neuron 56 (2): 327-338.

  • Neuroscience Research. Windhorst U and Johansson H (Editors) Springer Verlag, pp 893-969.

  • Hubener M, Bonhoeffer T. Visual cortex: Two-photon excitement Current Biology 15 (6): R205-R208 mar 29 2005

  • Grinvald A, Hildesheim R. VSDI: A new era in functional imaging of cortical dynamics. Nature Reviews Neuroscience 5 (11): 874-885 nov 2004

  • Zapeda A, Arias C, Sengpiel F. Optical imaging of intrinsic signals: recent developments in the methodology and its applications. Journal Of Neuroscience Methods 136 (1): 1-21 jun 15 2004

  • Grinvald A, et al. (1999). In-vivo Optical Imaging of cortical Architecture and Dynamics. In Modern Techniques in Neuroscience Research. U. Windhorst and H. Johansson Springer, pp 893-969

  • Mrsic-Flogel T, Hubener M, Bonhoeffer T. Brain mapping: New wave optical Imaging. C urrent B iology 13 (19): R778-R780 SEP 30 2003

  • Grinvald A (1985). Real-time optical mapping of neuronal activity: from single growth cones to the intact mammalian brain. Ann Rev Neurosci, 8: 263-305.