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, 7 (9), 3280-3288
eCollection

Microdose Fluorescence Imaging of ABY-029 on an Operating Microscope Adapted by Custom Illumination and Imaging Modules

Affiliations

Microdose Fluorescence Imaging of ABY-029 on an Operating Microscope Adapted by Custom Illumination and Imaging Modules

Jonathan T Elliott et al. Biomed Opt Express.

Abstract

Fluorescence guided surgery has the potential to positively impact surgical oncology; current operating microscopes and stand-alone imaging systems are too insensitive or too cumbersome to maximally take advantage of new tumor-specific agents developed through the microdose pathway. To this end, a custom-built illumination and imaging module enabling picomolar-sensitive near-infrared fluorescence imaging on a commercial operating microscope is described. The limits of detection and system specifications are characterized, and in vivo efficacy of the system in detecting ABY-029 is evaluated in a rat orthotopic glioma model following microdose injections, showing the suitability of the device for microdose phase 0 clinical trials.

Keywords: (170.1610) Clinical applications; (170.3880) Medical and biological imaging; (170.4730) Optical pathology; (260.2510) Fluorescence.

Figures

Fig. 1
Fig. 1
The detection module (blue) and the illumination model (yellow) are attached to the Zeiss Pentero OPMI head and can be used without affecting standard operation of the microscope.
Fig. 2
Fig. 2
(A) The laser (black line) is cleaned up by the ET770spuv shortpass filter before exiting the illumination module. This excitation along with coincident white-light from the Pentero is reflected by the T770lpxr dichroic towards the RGB camera. The fluorescence emission is transmitted through the dichroic and filtered by the ET780lp long-pass filter before sCMOS detection. (B) Maximum laser beam power, measured in the center of the illumination field was characterized using a power meter affixed with a 7-mm diameter aperture at different distances from the bottom of the adapter. The green shaded area indicates power per area below the ANSI limit for skin exposure for a continuous-wave 760 nm light source. The blue shaded area shows the typical working distance used during surgery. The IEC 3R Laser class limit is represented by the dashed grey line.
Fig. 3
Fig. 3
The relationship between signal and ABY-029 concentration for each system plotted on a log-log scale. When evaluating the microdose module, the Pentero xenon arc lamp was either off (squares) or on at 30% power (dots). Dashed lines show the line-of-best-fit over the linear response regions, and dotted lines show the noise floor. Black crosses show the intersection point which defines the lower limit of detection.
Fig. 4
Fig. 4
(A) The overlay (fusion) of two images acquired simultaneously during open craniotomy with the custom imaging module: the RGB white-light image and the fluorescence image overlay 2-h following a 3x microdose injection. (B) The ex vivo overlay of the whole brain acquired 2-h following a 6x microdose injection. (C) The corresponding ex vivo fluorescence image acquired with the Pentero IR800 channel. (D) A PpIX image acquired with the Zeiss Pentero BLUE400 channel, and (E) the same brain imaged by the custom imaging module during blue-light excitation. (F) The same brain as in D and E, imaged for ABY-029 fluorescence using the customized Pentero 2-h following a 3x microdose injection.

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