Autofluorescence lifetime augmented reality as a means for real-time robotic surgery guidance in human patients

Due to loss of tactile feedback the assessment of tumor margins during robotic surgery is based only on visual inspection, which is neither significantly sensitive nor specific. Here we demonstrate time-resolved fluorescence spectroscopy (TRFS) as a novel technique to complement the visual inspection of oral cancers during transoral robotic surgery (TORS) in real-time and without the need for exogenous contrast agents. TRFS enables identification of cancerous tissue by its distinct autofluorescence signature that is associated with the alteration of tissue structure and biochemical profile. A prototype TRFS instrument was integrated synergistically with the da Vinci Surgical robot and the combined system was validated in swine and human patients. Label-free and real-time assessment and visualization of tissue biochemical features during robotic surgery procedure, as demonstrated here, not only has the potential to improve the intraoperative decision making during TORS but also other robotic procedures without modification of conventional clinical protocols

Reply to Comment: ‘A novel method for fast and robust estimation of fluorescence decay dynamics using constrained least-square deconvolution with Laguerre expansion’

In this response we underscore that the instrumentation described in the original publication (Liu et al 2012 Phys. Med. Biol. 57 843-65) was based on pulse-sampling technique, while the comment by Zhang et al is based on the assumption that a time-correlated single photon counting (TCSPC) instrumentation was used. Therefore the arguments made in the comment are not applicable to the noise model reported by Liu et al. As reported in the literature (Lakowicz 2006 Principles of Fluorescence Spectroscopy (New York: Springer)), while in the TCSPC the experimental noise can be estimated from Poisson statistics, such an assumption is not valid for pulse-sampling (transient recording) techniques. To further clarify this aspect, we present here a comprehensive noise model describing the signal and noise propagation of the pulse sampling time-resolved fluorescence detection. Experimental data recorded in various conditions are analyzed as a case study to demonstrate the noise model of our instrumental system. In addition, regarding the statement of correcting equation (3) in Liu et al (2012 Phys. Med. Biol. 57 843-65), the notation of discrete time Laguerre function in the original publication was clear and consistent with literature conventions (Marmarelis 1993 Ann. Biomed. Eng. 21 573-89, Westwick and Kearney 2003 Identification of Nonlinear Physiological Systems (Hoboken, NJ: Wiley)). Thus, it does not require revision

Technique for real-time tissue characterization based on scanning multispectral fluorescence lifetime spectroscopy (ms-TRFS)

We report a novel technique for continuous acquisition, processing and display of fluorescence lifetimes enabling real-time tissue diagnosis through a single hand held or biopsy fiber-optic probe. A scanning multispectral time-resolved fluorescence spectroscopy (ms-TRFS) with self-adjustable photon detection range was developed to account for the dynamic changes of fluorescence intensity typically encountered in clinical application. A fast algorithm was implemented in the ms-TRFS software platform, providing up to 15 Hz continuous display of fluorescence lifetime values. Potential applications of this technique, including biopsy guidance, and surgical margins delineation were demonstrated in proof-of-concept experiments. Current results showed accurate display of fluorescence lifetimes values and discrimination of distinct fluorescence markers and tissue types in real-time (< 100 ms per data point)

Real-Time Visualization of Tissue Surface Biochemical Features Derived from Fluorescence Lifetime Measurements

Fiber based fluorescence lifetime imaging has shown great potential for intraoperative diagnosis and guidance of surgical procedures. Here we describe a novel method addressing a significant challenge for the practical implementation of this technique, i.e., the real-time display of the quantified biochemical or functional tissue properties superimposed on the interrogated area. Specifically, an aiming beam (450 nm) generated by a continuous-wave laser beam was merged with the pulsed fluorescence excitation light in a single delivery/collection fiber and then imaged and segmented using a color-based algorithm. We demonstrate that this approach enables continuous delineation of the interrogated location and dynamic augmentation of the acquired frames with the corresponding fluorescence decay parameters. The method was evaluated on a fluorescence phantom and fresh tissue samples. Current results demonstrate that 34 frames per second can be achieved for augmenting videos of 640 × 512 pixels resolution. Also we show that the spatial resolution of the fluorescence lifetime map depends on the tissue optical properties, the scanning speed, and the frame rate. The dice similarity coefficient between the fluorescence phantom and the reconstructed maps was estimated to be as high as 93%. The reported method could become a valuable tool for augmenting the surgeon's field of view with diagnostic information derived from the analysis of fluorescence lifetime data in real-time using handheld, automated, or endoscopic scanning systems. Current method provides also a means for maintaining the tissue light exposure within safety limits. This study provides a framework for using an aiming beam with other point spectroscopy applications

Real-Time Visualization of Tissue Surface Biochemical Features Derived From Fluorescence Lifetime Measurements

Fiber based fluorescence lifetime imaging has shown great potential for intraoperative diagnosis and guidance of surgical procedures. Here we describe a novel method addressing a significant challenge for the practical implementation of this technique, i.e. the real-time display of the quantified biochemical or functional tissue properties superimposed on the interrogated area. Specifically, an aiming beam (450 nm) generated by a continuous-wave laser beam was merged with the pulsed fluorescence excitation light in a single delivery/collection fiber and then imaged and segmented using a color-based algorithm. We demonstrate that this approach enables continuous delineation of the interrogated location and dynamic augmentation of the acquired frames with the corresponding fluorescence decay parameters. The method was evaluated on a fluorescence phantom and fresh tissue samples. Current results demonstrate that 34 frames per second can be achieved for augmenting videos of 640×512 pixels resolution. Also we show that the spatial resolution of the fluorescence lifetime map depends on the tissue optical properties, the scanning speed, and the frame rate. The dice similarity coefficient between the fluorescence phantom and the reconstructed maps was estimated to be as high as 93%. The reported method could become a valuable tool for augmenting the surgeon’s field of view with diagnostic information derived from the analysis of fluorescence lifetime data in real-time using handheld, automated, or endoscopic scanning systems. Current method provides also a means for maintaining the tissue light exposure within safety limits. This study provides a framework for using an aiming beam with other point spectroscopy applications

Reply to Comment: ‘A novel method for fast and robust estimation of fluorescence decay dynamics using constrained least-square deconvolution with Laguerre expansion’

In this response we underscore that the instrumentation described in the original publication (Liu et al 2012 Phys. Med. Biol. 57 843-65) was based on pulse-sampling technique, while the comment by Zhang et al is based on the assumption that a time-correlated single photon counting (TCSPC) instrumentation was used. Therefore the arguments made in the comment are not applicable to the noise model reported by Liu et al. As reported in the literature (Lakowicz 2006 Principles of Fluorescence Spectroscopy (New York: Springer)), while in the TCSPC the experimental noise can be estimated from Poisson statistics, such an assumption is not valid for pulse-sampling (transient recording) techniques. To further clarify this aspect, we present here a comprehensive noise model describing the signal and noise propagation of the pulse sampling time-resolved fluorescence detection. Experimental data recorded in various conditions are analyzed as a case study to demonstrate the noise model of our instrumental system. In addition, regarding the statement of correcting equation (3) in Liu et al (2012 Phys. Med. Biol. 57 843-65), the notation of discrete time Laguerre function in the original publication was clear and consistent with literature conventions (Marmarelis 1993 Ann. Biomed. Eng. 21 573-89, Westwick and Kearney 2003 Identification of Nonlinear Physiological Systems (Hoboken, NJ: Wiley)). Thus, it does not require revision

Rotational multispectral fluorescence lifetime imaging and intravascular ultrasound: bimodal system for intravascular applications

We report the development and validation of a hybrid intravascular diagnostic system combining multispectral fluorescence lifetime imaging (FLIm) and intravascular ultrasound (IVUS) for cardiovascular imaging applications. A prototype FLIm system based on fluorescence pulse sampling technique providing information on artery biochemical composition was integrated with a commercial IVUS system providing information on artery morphology. A customized 3-Fr bimodal catheter combining a rotational side-view fiberoptic and a 40-MHz IVUS transducer was constructed for sequential helical scanning (rotation and pullback) of tubular structures. Validation of this bimodal approach was conducted in pig heart coronary arteries. Spatial resolution, fluorescence detection efficiency, pulse broadening effect, and lifetime measurement variability of the FLIm system were systematically evaluated. Current results show that this system is capable of temporarily resolving the fluorescence emission simultaneously in multiple spectral channels in a single pullback sequence. Accurate measurements of fluorescence decay characteristics from arterial segments can be obtained rapidly (e.g., 20 mm in 5 s), and accurate co-registration of fluorescence and ultrasound features can be achieved. The current finding demonstrates the compatibility of FLIm instrumentation with in vivo clinical investigations and its potential to complement conventional IVUS during catheterization procedures