Fluorescence-enhanced optical imaging in large tissue volumes using a gain-modulated ICCD camera.

A novel image-intensified charge-coupled device (ICCD) imaging system has been developed to perform 3D fluorescence tomographic imaging in the frequency-domain using near-infrared contrast agents. The imager is unique since it (i) employs a large tissue-mimicking phantom, which is shaped and sized to resemble a female breast and part of the extended chest-wall region, and (ii) enables rapid data acquisition in the frequency-domain by using a gain-modulated ICCD camera. Diffusion model predictions are compared to experimental measurements using two different referencing schemes under two different experimental conditions of perfect and imperfect uptake of fluorescent agent into a target. From these experimental measurements, three-dimensional images of fluorescent absorption were reconstructed using a computationally efficient variant of the approximate extended Kalman filter algorithm. The current work represents the first time that 3D fluorescence-enhanced optical tomographic reconstructions have been achieved from experimental measurements of the time-dependent light propagation on a clinically relevant breast-shaped tissue phantom using a gain-modulated ICCD camera.

Near-infrared fluorescence contrast-enhanced imaging with area illumination and area detection: the forward imaging problem.

Fluorescence frequency-domain photon migration measurements were acquired from tissue phantoms, each containing a fluorescent target, by means of area illumination and area detection on the same surface and for the first time, to our knowledge, compared with predictions computed with a numerical solution to the coupled photon diffusion equations. We accomplished area illumination and area detection using a planar, intensity-modulated excitation light source and a gain-modulated intensified charge-coupled device camera, respectively. A 1-ml vessel containing 1-microm solution of Indocyanine Green in 1% Liposyn was immersed 1 cm deep in each 512-ml tissue phantom. For most tissue phantoms, the background surrounding the 1-ml target was composed of Liposyn solution containing Indocyanine Green or 3,3'-Diethylthiatricarbocyanine Iodide such that the target-to-background ratio of fluorescence yield was > or = 10:1. Measurements of fluorescence modulation amplitude and phase were predicted with a mean error ranging from 10.1% to 13.6% and 0.56 degrees to 1.72 degrees, respectively. These numbers are similar to those obtained by use of single-pixel frequency-domain photon migration techniques and validate the potential use of area illumination and area detection for biomedical imaging of tissues. Results also demonstrate that target-to-background ratios of fluorescence yield and fluorescence lifetime significantly affect target detectability.

Sensitivity and depth penetration of continuous wave versus frequency-domain photon migration near-infrared fluorescence contrast-enhanced imaging.

The development of near-infrared fluorescent contrast agents and imaging techniques depends on the deep penetration of excitation light through several centimeters of tissue and the sensitive collection of the re-emitted fluorescence. In this contribution, the sensitivity and depth penetration of various fluorescence-enhanced imaging studies is surveyed and compared with current studies using continuous wave (CW) and frequency-domain photon migration (FDPM) measurements with planar wave illumination of modulated excitation light at 100 MHz and area collection of reemitted fluorescent light using a previously developed modulated intensified charge-coupled device camera system. Fluorescence was generated from nanomolar to micromolar solutions of indocyanine green (ICG) in a 100 microL volume submerged at 1-4 cm depths in a 1% Liposyn solution to mimic tissue scattering properties. Enhanced depth penetration and sensitivity are achieved with optimal filter rejection of excitation light, and FDPM rejection of background light is not achieved using CW methods. We show the ability to detect as few as 100 fmol of ICG from area illumination of 785 nm light (5.5 mW/cm2) and FDPM area collection of 830 nm fluorescent light generated from 3 cm below the phantom surface. The lowered noise floor of FDPM measurements enables greater sensitivity and penetration depth than comparable CW measurements.

Near-infrared fluorescence contrast-enhanced imaging with intensified charge-coupled device homodyne detection: measurement precision and accuracy.

Fluorescence frequency-domain photon migration (FDPM) through tissue refers to the propagation of intensity-modulated fluorescent light that originates from tissue-laden fluorophores following illumination with an intensity-modulated excitation light source. FDPM measurements of modulation amplitude and phase are ultimately employed in an inversion algorithm for tomographic reconstruction of interior optical and fluorescent property maps that delineate disease enhanced with fluorescent contrast agent. Because the inverse problem is underdetermined, measurement precision and accuracy crucially impact its solution. Reported here are the precision and accuracy of FDPM measurements acquired using an intensified CCD homodyne detection system. By introducing 32 phase delays between the oscillators used to modulate the intensifier gain and light source intensity at 100 MHz, mean precision is maximized at +/-0.46% and +/-0.26 deg for measurements of modulation amplitude and phase, respectively. Measurement precision improves when the number of phase delays increases. Measurements of fluorescence modulation amplitude and phase, acquired from the surface of a tissue phantom at distances ranging between 0.71 and 3.6 cm from an incident excitation point source, exhibit a mean accuracy of 17% and 1.9 deg, respectively. Measurement accuracy deteriorates with increasing distance from the point source, but for distances up to 1.0 cm from the point source, measurements of fluorescence modulation amplitude and phase exhibit a mean accuracy of 5.4% and 0.30 deg, respectively.