Time-resolved scanning system for double reflectance and transmittance fluorescence imaging of diffusive media.
In this work we present a novel diffuse fluorescence imaging system, based on time-resolved two-wavelength double reflectance and transmittance setup for slab geometry samples. We describe the hardware setup, showing its compactness and versatility and show the results on preliminary measurements on phantoms. We fully assessed the performances and the dynamic ranges of the system. We validated its ability of recovering the optical properties of the bulk medium, for samples with scattering and absorption coefficients similar to those of biological tissues and with thicknesses of about 2 cm. Moreover we assess the linearity of the recorded signals against the fluorophore concentration, when it is homogeneously diffused in the phantom or concentrated inside a sealed inclusion. In both cases we observe again a fairly good linearity, over three orders of magnitude, from 10(-8)M to 10(-5)M. With the fluorescent inclusion we were also able to assess the imaging capabilities of the system, in terms of spatial resolution, which we appraise in about 3 mm, and in terms of imaging sensitivity (the smallest quantity of fluorescent dye distinguishable from the homogeneous background), settled to 200 fmol. Since the recorded data are time resolved, we could also estimate the dye fluorescence lifetime and build early and late time gate images. We finally discuss some of the criticalities of the proposed system and the developments we are currently carrying on in order to adapt it for in vivo measurements.
Brambilla
Marco
M
IIT, ULTRAS-INFM-CNR and IFN-CNR, Politecnico di Milano, Dipartimento di Fisica, Piazza Leonardo da Vinci 32, I-20133 Milan, Italy. m.brambilla@polimi.it
Spinelli
Lorenzo
L
Pifferi
Antonio
A
Torricelli
Alessandro
A
Cubeddu
Rinaldo
R
D004058:Diffusion; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation* / Q000379:methods; D018613:Microscopy, Confocal / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D009391:Nephelometry and Turbidimetry / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014054:Tomography / Q000295:instrumentation* / Q000379:methods