Human bladder cancer diagnosis using Multiphoton microscopy
ABSTRACT At the time of diagnosis, approximately 75% of bladder cancers are non-muscle invasive. Appropriate diagnosis and surgical resection at this stage improves prognosis dramatically. However, these lesions, being small and/or flat, are often missed by conventional white-light cystoscopes. Furthermore, it is difficult to assess the surgical margin for negativity using conventional cystoscopes. Resultantly, the recurrence rates in patients with early bladder cancer are very high. This is currently addressed by repeat cystoscopies and biopsies, which can last throughout the life of a patient, increasing cost and patient morbidity. Multiphoton endoscopes offer a potential solution, allowing real time, non-invasive biopsies of the human bladder, as well as an up-close assessment of the resection margin. While miniaturization of the Multiphoton microscope into an endoscopic format is currently in progress, we present results here indicating that Multiphoton imaging (using a bench-top Multiphoton microscope) can indeed identify cancers in fresh, unfixed human bladder biopsies. Multiphoton images are acquired in two channels: (1) broadband autofluorescence from cells, and (2) second harmonic generation (SHG), mostly by tissue collagen. These images are then compared with gold standard hematoxylin/eosin (H&E) stained histopathology slides from the same specimen. Based on a "training set" and a very small "blinded set" of samples, we have found excellent correlation between the Multiphoton and histopathological diagnoses. A larger blinded analysis by two independent uropathologists is currently in progress. We expect that the conclusion of this phase will provide us with diagnostic accuracy estimates, as well as the degree of inter-observer heterogeneity.
Full-textDOI: · Available from: Mohammed Akhtar, May 06, 2015
Click to see the full-text of:
Article: Human bladder cancer diagnosis using Multiphoton microscopy
SourceAvailable from: Henricus J C M Sterenborg[Show abstract] [Hide abstract]
ABSTRACT: Nonlinear optical imaging (NLOI) has emerged to be a promising tool for bio-medical imaging in recent times. Among the various applications of NLOI, its utility is the most significant in the field of pre-clinical and clinical cancer research. This review begins by briefly covering the core principles involved in NLOI, such as two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG). Subsequently, there is a short description on the various cellular components that contribute to endogenous optical fluorescence. Later on the review deals with its main theme – the challenges faced during label-free NLO imaging in translational cancer research. While this review addresses the accomplishment of various label-free NLOI based studies in cancer diagnostics, it also touches upon the limitations of the mentioned studies. In addition, areas in cancer research that need to be further investigated by label-free NLOI are discussed in a latter segment. The review eventually concludes on the note that label-free NLOI has and will continue to contribute richly in translational cancer research, to eventually provide a very reliable, yet minimally invasive cancer diagnostic tool for the patient.Journal of photochemistry and photobiology. B, Biology 09/2014; 141(2014):128-138. DOI:10.1016/j.jphotobiol.2014.08.025 · 2.80 Impact Factor
[Show abstract] [Hide abstract]
ABSTRACT: Bladder cancer presents a spectrum of different diatheses. A precise assessment for individualized treatment depends on the accuracy of the initial diagnosis. Detection relies on comprehensive and accurate white light cystoscopy. White light cystoscopy has limitations in addition to its invasive nature and the potential risks related to the method. These limitations include difficulties in flat lesion detection, precise tumor delineation to enable complete resection, inflammation and malignancy differentiation, and grade and stage determination. The resolution of these problems depends on the surgeon’s ability and experience with available technology for visualization and resection. In this study, we used multi-spectral imaging technology combined with phase contrast microscopy to analyze bladder cancer cells (BCCs) at various stages using a single-cell array chip. We found from the spectral characteristics of single cell that the cell spectra at the different cancer stages demonstrate a change in the cell’s composition. We cultured 419 normal and diseased bladder cells (BCs). We used principal component analysis and a principal component score map to distinguish the different cancer stages. Diagnosis sensitivity and specificity of this method were 85.7% and 90.2% in 119 stage 0 (normal) cells, 84.3% and 90.8% in 79 stage 2 cancer cells, 87.6% and 92.4% in 151 stage 3 BCCs, and 85.3% and 91.2% in 70 stage 4 BCCs, respectively.IEEE Journal of Selected Topics in Quantum Electronics 05/2014; 20(3):68800808. DOI:10.1109/JSTQE.2013.2279804 · 3.47 Impact Factor
[Show abstract] [Hide abstract]
ABSTRACT: Multiphoton microscopy (MPM) enables real-time imaging of various cellular processes at submicron resolution. MPM is currently being used in neuroscience, oncology, and immunology. MPM has demonstrated promising results in urology. MPM has been used in the identification of spermatogenesis, evaluation of bladder cancer, and tissue identification in prostate cancer surgery. MPM has allowed the visualization of seminiferous tubules within the testis in a rat model and identified areas of spermatogenesis. MPM could potentially improve the efficacy of testicular sperm extraction. In bladder cancer evaluation, MPM has proven to be an effective imaging tool in identifying areas suspicious for malignancy. The imaging technology could be utilized in the future to provide urologists with an immediate impression of extracted bladder tissue, or as part of a cystoscopic device to evaluate the bladder in real time. Similarly, MPM has proven to be a useful imaging technique to evaluate prostate cancer. MPM could be utilized during a prostatectomy to help differentiate prostate from cavernous nerves that are closely adherent to the prostate. MPM uses a laser and safety studies will need to be performed prior to its utilization in the clinical setting.