As we know Quantum mechanics is a mathematical theory that can describe the behavior of objects that are at
microscopic level. Time-resolved autofluorescence spectrometer monitors events that occur during the lifetime of the
excited state. This time ranges from a few picoseconds to hundreds of nanoseconds. That is an extremely important
advance as it allows environmental parameters to be monitored in a spatially defined manner in the specimen under
study. This technique is based on the application of Quantum Mechanics. This principle is applied in our project as we
are trying to use different fluorescence spectra to detect biological molecules commonly found in cancerous colorectal
tissue and thereby differentiate the cancerous and non-cancerous colorectal polyps more accurately and specifically. In
this paper, we use Fluorescence Lifetime Spectrometer (Edinburgh Instruments FL920) to measure decay time of
autofluorescence of colorectal cancerous and normal tissue sample. All specimens are from Department of Colorectal
Surgery, Singapore General Hospital. The tissues are placed in the time-resolved autofluorescence instrument, which
records and calculates the decay time of the autofluorescence in the tissue sample at the excitation and emission
wavelengths pre-determined from a conventional spectrometer. By studying the decay time,τ, etc. for cancerous and
normal tissue, we aim to present time-resolved autofluorescence as a feasible technique for earlier detection of malignant
colorectal tissues. By using this concept, we try to contribute an algorithm even an application tool for real time early
diagnosis of colorectal cancer for clinical services.
This study aimed at applying Laser induced-autofluorescence (LIAF) diagnostics method as an in-vivo screening of colorectal polyplcancer. The spectrum algorithm based on the ratio of autofluorescence intensity was used to identify the diseased tissues from the normal tissues as it was generally performed better than an algorithm based only simply on the intensity of the spectrum. Histopathological biopsy results were compared with the detected AF spectra characteristics for different kinds of polyps. 73 patients had been examined via the LIAF spectroscopy detection system during their colonoscopy screening in Endoscopy Center, Singapore General Hospital. The autofluorescence from the surface of the colorectal tissues under 405 nm laser light excitation was detected using our detecting system. In the experimental investigation two groups of patients were involved. One group was "abnormal" group. There were 25 patients belonging to this group since polyps or carcinoma was found in their colorectal tract during colonoscopy. The histopathology reports confirm the group classification. Total 36 polyps' AF spectra and 9 carcinoma' AF spectra were detected from 25
patients of the abnormal group during their regular endoscopy examination. The intensity ratios RI-680/I-500 and RI-630/I-500 of polyps/cancerous AF spectra and intensity ratios of corresponding normal colorectal AF spectra were calculated. Two
critical intensity ratios for separating the AF intensity ratios RI-680/I-500 and RI-630/I-500 of normal and abnormal colorectal tissues were defined as 0.5 and 0.6 respectively. Using the critical intensity ratio values, 48 "normal" group patients' rectums were checked via the LIAF detection system. There were 20 patients (41.7%) whose AF spectra of colorectal tract mucosa belonging to abnormal spectra. However, these 20 patients had not been found under white light via traditional endoscopy. For small diseased area like small plat polyp disease and carcinoma, it was very difficult to identify under white light by endoscopy. However, the LIAF spectra technique and AF intensity ratio algorithm was able
to detect these kinds of abnormal area earlier than traditional endoscopy. Using this algorithm, it is able to identify the onset of abnormal tissue growth during real-time clinical endoscope examination.
We investigated normal and cancerous human colorectal tissues (fresh thick biopsy specimens) using Olympus Confocal laser scanning biological microscope (FV300). The different layers of autofluorescence images of the specimen were captured by 488 nm laser scanning and sectioning. Optical sectioning can be performed in the vertical plane. Laser scanning can be performed in the horizontal plane. By comparing the autofluorescence image of the normal colorectal tissue with cancerous tissue, the structures of the optical sectioning image layer were found to be significantly different. We have also obtained fibrous autofluorescence image inside tissue specimen. Our investigation may help provide some useful insight to other autofluorescence research studies like laser induced autofluorescence spectra of human colorectal tissue study as a diagnosis technique for clinical application.
We investigated 48 normal patients and 25 diseased patients using our laser-induced autofluorescence spectra detection system during their regular colonoscopy. The colon and rectum mucosa autofluorescence were excited by 405 nm continue wavelength laser. We observed that cancer or diseased colorectal mucosa, their autofluorescence spectra are significantly different from normal area. The autofluorescence spectra intensity at about 500 nm was been used for our intensity ratio characteristics intensity for our diagnostic algorithm. The intensity ratios of RI-680/I-500 and RI-630/I-500 were performed to identify the detection area. From experimental result we concluded that both intensity ratios of RI-680/I-500 and RI-630/I-500 as guidelines can detect cancerous and polyps disease completely. Our investigation provided some useful insight for laser induced autofluorescence spectra as a diagnosis technique for clinical application.
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