The experimental noncontact FMT setup used [Fig. 1(a)] employed a continuous-wave argon-ion laser source (LaserPhysics, Reliant 1000m, West Jordan, Utah) emitting at 457, 488, and 514 nm and enclosed in a custom-built aluminum chamber coated in antireflective black coating (deep black optical paint, Gerd Neumann, Hamburg, Germany). The source was directed across three fixed mirrors (High Energy Argon-Ion Laser Mirrors, Edmund Optics, Barrington, Illinois) and a UV–NIR neutral density filter (Edmund Optics) into the FMT chamber and onto an optical scanner employing galvo-mirrors (Scanlab, SCANcube 7, Munich, Germany). Incident light power was measured before entering the optical scanner using a laser power meter (PH100-Si-OD2, Gentec Electro-Optics). The setup was designed to operate in both transmission and reflection geometries by repositioning a mirror (First Surface mirror, Edmund Optics) mounted on a travel linear translation stage (Edmund Optics). The sample holder was mounted on an electronically controlled positioning stage (Standa, Vilnius, Lithuania). A custom-built filter wheel accommodating up to five filters was aligned to a 50 mm Macro objective (SIGMA Corporation, Tokyo, Japan). Light collection was performed using a -, -, -, or interference bandpass filters (Andover Corporation, Salem, New Hampshire) to isolate fluorescence from excitation light. A 16-bit, charge-coupled device (CCD) (DV 434, Andor Technology, Belfast, Northern Ireland), thermoelectrically cooled down to , was used for the detection of signals. The lens was focused on the surface of the sample, and the image scaled from pixel dimensions to metric units by the use of metric fiducials. Stage calibration was performed to convert the CCD field of view to degrees of rotation of the galvo-mirrors. For all experiments, an image was obtained with white light illumination before exposure to the laser source and was used as a reference image for source pattern selection. Additionally, a background image in the absence of an illuminating source was acquired to account for any external light entering the FMT chamber. Illumination sources were distributed symmetrically to the target location for both reproducibility and calibration experiments. Exposure time used was 0.2 s with 0.8 mW excitation power. Identical source/detector parameters were used for the fluorophores Atto590 (excitation max 594 nm, emission max 624 nm; ATTO-TEC Gmbh, Germany) and carboxyfluorescein succidimyl ester (CFSE) (excitation max 492 nm, emission max 517 nm; Thermo Fisher Scientific Inc.) placed in a 2-mm capillary glass tube in a phantom at a depth of 3 mm from the surface. A slab geometry phantom was used with a combination of 20 ppm India ink and 1% Intralipid-20% to match tissue-like optical properties at 520 nm (scattering coefficient and absorption coefficient ) and at a depth of 12 mm. An isoflurane vaporizer was used to deliver anesthetic to the FMT chamber and to the induction chamber, and a scavenging system was used to remove excess anesthetic gas (Fluovac systems, Harvard Apparatus, Hollison, Massachusetts).