Light-induced fluorescence spectroscopy of adenomas, adenocarcinomas and non-neoplastic mucosa in human colon I. In vitro measurements

doi:10.1016/1011-1344(92)85100-9

Journal of Photochemistry and Photobiology B: Biology

Volume 14, Issue 3, 15 July 1992, Pages 219-230

https://doi.org/10.1016/1011-1344(92)85100-9 Get rights and content

Abstract

In an attempt to evaluate whether induced fluorescence could be exploited to discriminate neoplastic from non-neoplastic tissue, fluorescence spectroscopy was performed at 450–800 nm on 83 biopsy specimens of colonic mucosa. Measurements showed that fluorescence spectra of adenoma, adenocarcinoma and non-neoplastic mucosa manifest dissimilar patterns. Nine variables, whose photophysical and/or biological bases need further investigation, were derived from the spectra. Discriminant functions between the groups of lesions were determined by using a stepwise discriminant analysis. The diagnostic test had a sensitivity of 80.6% and 88.2%, and a specificity of 90.5% and 95.2% in discriminating neoplastic from non-neoplastic mucosa and adenoma from non-neoplastic mucosa respectively. These results suggest that fluorescence spectroscopy has the potential to improve endoscopic diagnosis of premalignant and malignant lesions of colonic mucosa.

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An effective approach to the early diagnosis of colorectal cancer based on three-dimensional fluorescence spectra of human blood plasma
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When excited at a certain wavelength, these tissues would produce a wide fluorescence emission spectrum in the UV–vis regions, attributing to the collagen, elastin, reduced form of nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD) and porphyrins [11]. Previous studies indicated that the fluorescence spectra of adenomas, adenocarcinomas and non-neoplastic mucosa in human colon showed significant differences [12]. With the development of histopathology, the changes of the biochemical and physicochemical properties of cells and tissues could be observed according to the changes of fluorescence emission spectra.
Colorectal cancer (CRC) is a common malignancy in the gastrointestinal tract, and its screening rates remain relatively low in the general population due to the lack of specific symptoms and effective methods. It is still in urgent need to develop rapid and reliable approach to the early diagnosis of CRC. Herein, based on the three-dimensional (3D) fluorescence spectra of human blood plasma, a combination strategy of Tchebichef image moments coupled with partial least squares-discriminate analysis (TM-PLS-DA) was proposed for the detection of CRC from three classes (CRC samples, adenomas samples and non-malignant findings). The established TM-PLS-DA classification model provided an 84 % correct classification for CRC prediction. Venetian blinds 10-fold cross validation was carried out. The error rates both in cross validation and test sets were less than 0.16. Sensitivity and specificity for CRC prediction were 0.95 and 0.88, respectively. At the same time, the diagnostic capacity of the proposed method was tested by receiver operating characteristics (ROC) analysis with area under the curve (AUC) of 0.94 for CRC diagnosis. These results demonstrate that the proposed TM-PLS-DA method based on the 3D fluorescence spectra of blood plasma has great advantage for the accurate CRC detection, which will provide a potential alternative approach for cancer diagnostics.
Multimodal hyperspectroscopy as a triage test for cervical neoplasia: Pivotal clinical trial results
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Citation Excerpt :
The primary goal of this study was to provide a prospective evaluation of the sensitivity and specificity of MHS for the detection of moderate and high grade dysplasia and, using this information, provides insights for how this new test could be used to improve care for women at risk for these conditions. Tissue spectroscopy has been evaluated in many clinical trials for detecting neoplasia of the cervix [3–7], lung [8], gastrointestinal tract [9–11], and skin [12]. These earlier systems typically utilized a single excitation wavelength or a single spectroscopic mode [8,13–15].
To prospectively evaluate a new non invasive device that combines fluorescence and reflectance spectroscopy in a population in women at risk for cervical dysplasia.
A total of 1607 women were evaluated with multimodal hyperspectroscopy (MHS), a painless test with extremely high spectral resolution. Subjects who were referred to colposcopy based on abnormal screening tests or other referral criteria underwent the MHS test and also had a sample taken for additional cytology and presence of high risk human papilloma virus (HPV) prior to undergoing biopsy.
Sensitivity of MHS for cervical intraepithelial neoplasia (CIN) 2 + was 91.3% (252/276). Specificity, or the potential reduction in referrals to colposcopy and biopsy, was 38.9% (222/570) for women with normal or benign histology and 30.3% (182/601) for women with CIN1 histology. Two year follow-up data, collected for a subgroup of 804 women, revealed 67 interval CIN2 + that originally were diagnosed at enrollment as normal or CIN1. MHS identified 60 of these (89.6%) as positive for CIN2 + prior to their discovery during the two year follow-up period.
MHS provides an immediate result at the point of care. Recently, the limitations of cytology have become more obvious and as a consequence greater emphasis is being placed on HPV testing for cervical cancer screening, creating a need for an inexpensive, convenient and accurate test to reduce false positive referrals to colposcopy and increase the yield of CIN2 + at biopsy. MHS appears to have many of the attributes necessary for such an application.
Pattern recognition of multiple excitation autofluorescence spectra for colon tissue classification
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The aim of this study was to explore the usefulness of multiple excitation autofluorescence (AF) and a spectral feature-based pattern recognition in classification of colon tissues.
Under four different excitation wavelengths (337, 375, 405 and 460 nm), AF spectra of freshly excised normal and adenocarcinoma colon tissues were measured. Pattern recognition method including features extraction, data reduction using principal component analysis (PCA) and Fisher's discriminant analysis (FDA) were performed for classification.
There was a significantly difference between spectral patterns of normal and adenocarcinoma tissues. Compared with the other three excitation wavelengths, the AF spectra obtained under 337 nm excitation provided more diagnostic information, but also more sensitive to the trivial change resulted from neoplastic transformation. For discriminating normal from adenocarcinoma tissues, the sensitivity, specificity and accuracy using 337 nm excitation in the present study were 88.9%, 80.0% and 83.9%, respectively. Compared these values with those determined from multispectral data analysis, our findings indicate that the latter has higher specificity while maintaining the same sensitivity (sensitivity 88.9% vs. 88.9%, specificity 91.4% vs. 80.0%, and accuracy 90.3% vs. 83.9%).
This study suggests that the pattern recognition of the multiple excitation AF spectra is an effective algorithm for improving the diagnostic accuracy of adenocarcinoma.
Numerical analysis of the autofluorescence intensity of neoplastic and non-neoplastic colorectal lesions by using a novel videoendoscopy system
2009, Gastrointestinal Endoscopy
Autofluorescence endoscopy (AFE) may improve detection and diagnosis of colorectal lesions. Recently, AFE based on a high-resolution video endoscope was developed.
A novel high-resolution video AFE system was used to quantify autofluorescence of colorectal lesions to determine the characteristics of non-neoplastic and neoplastic lesions.
Retrospective observational study.
Single-center referral hospital.
Ninety-seven patients with 103 colorectal lesions (22 non-neoplastic and 81 neoplastic lesions) who underwent AFE and were treated by using endoscopy or by surgery.
Recorded digital AFE images were analyzed to quantify autofluorescence. The following autofluorescence indexes were calculated: the green/red (G/R) ratio for each lesion, the color-contrast index between each lesion, and the corresponding normal region.
The G:R ratio, color-contrast index, and histopathologic characteristics for each colorectal lesion.
The mean G/R ratio was significantly higher in non-neoplastic lesions (1.17 [95% CI, 1.10-1.24], n = 22) than in neoplastic lesions (0.65 [95% CI, 0.63-0.68], n = 81) (P < .001). Mean color-contrast indexes were significantly lower in non-neoplastic lesions (7.99 [95% CI, 6.40-9.58], n = 22) than neoplastic lesions (35.06 [95% CI, 32.79-37.33], n = 81; P < .001). With a cutoff value of 1.01 for the G/R ratio and 13.94 for color-contrast index, AFE had a sensitivity and specificity of 98.8% and 86.4% respectively, for G/R ratio, and 98.8% and 90.9%, respectively, for color contrast index, in differentiating neoplastic from non-neoplastic colorectal lesions.
Retrospective design.
The quantification of digital AFE images obtained from the novel high-resolution videoendoscopy system revealed that autofluorescence was significantly different between non-neoplastic and neoplastic lesions, and color tone in AFE may represent the histopathologic characteristics of the lesion.
Multi-class classification algorithm for optical diagnosis of oral cancer
2006, Journal of Photochemistry and Photobiology B: Biology
We report development of a direct multi-class spectroscopic diagnostic algorithm for discrimination of high-grade cancerous tissue sites from low-grade as well as precancerous and normal squamous tissue sites of human oral cavity. The algorithm was developed making use of the recently formulated theory of total principal component regression (TPCR). The in vivo autofluorescence spectral data acquired from patients screened for neoplasm of oral cavity at the Government Cancer Hospital, Indore, was used to train and validate the algorithm. The diagnostic algorithm based on TPCR was found to provide satisfactory performance in classifying the tissue sites in four different classes – high-grade squamous cell carcinoma, low-grade squamous cell carcinoma, leukoplakia, and normal squamous tissue. The classification accuracy for these four classes was observed to be ∼94%, 100%, 100% and 91% for the training data set (based on leave-one-out cross-validation), and was ∼90%, 90%, 85% and 88%, respectively for the corresponding classes for the independent validation data set.
Cell and tissue autofluorescence research and diagnostic applications
2005, Biotechnology Annual Review
Citation Excerpt :
The DNF greatly improves the accuracy of diagnosis because the spectra are normalised with respect to their total integrated intensity, thus resulting independent from the intensity factor [79]. An in vitro study aimed to evaluate the efficacy of autofluorescence spectroscopy in colon cancer diagnosis demonstrated that fluorescence emission spectra, monitored in the range 450–800 nm, from adenoma, adenocarcinoma, and non-neoplastic tissues were significantly different [80]. In vitro analysis of normal colon tissues revealed autofluorescence signals mainly arising from fibres of the connective tissues (in particular from collagen) and eosinophils, present at the crypts level in the lamina propria of the mucosa, while colonic adenomas showed intensely fluorescent dysplastic epithelial cells in the crypts, a larger number of eosinophils and a decrease in autofluorescence emission due to connective component [81].
Cells contain molecules, which become fluorescent when excited by UV/Vis radiation of suitable wavelength. This fluorescence emission, arising from endogenous fluorophores, is an intrinsic property of cells and is called auto-fluorescence to be distinguished from fluorescent signals obtained by adding exogenous markers. The majority of cell auto-fluorescence originates from mitochondria and lysosomes. Together with aromatic amino acids and lipo-pigments, the most important endogenous fluorophores are pyridinic (NADPH) and flavin coenzymes. In tissues, the extracellular matrix often contributes to the auto-fluorescence emission more than the cellular component, because collagen and elastin have, among the endogenous fluorophores, a relatively high quantum yield. Changes occurring in the cell and tissue state during physiological and/or pathological processes result in modifications of the amount and distribution of endogenous fluorophores and chemical–physical properties of their microenvironment. Therefore, analytical techniques based on auto-fluorescence monitoring can be utilized in order to obtain information about morphological and physiological state of cells and tissues. Moreover, auto-fluorescence analysis can be performed in real time because it does not require any treatment of fixing or staining of the specimens. In the past few years spectroscopic and imaging techniques have been developed for many different applications both in basic research and diagnostics.

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Light-induced fluorescence spectroscopy of adenomas, adenocarcinomas and non-neoplastic mucosa in human colon I. In vitro measurements

Abstract

Am. Heart J.

Gastroenterology

Gastrointest. Endoscopy

Review of fluorescence diagnosis using porphyrins

SPIE

Fluorescence imaging and point measurements of tissue: applications to the demarcation of malignant tumors and atherosclerotic lesions from normal tissue

Photochem. Photobiol.

Behaviour of haematoporphyrin derivative in adenomas and adenocarcinomas of the colon: a microfluorimetric study

Lasers Med. Sci.

Human teeth with and without caries studied by laser scattering, fluorescence and absorption spectroscopy

IEEE Q.E.

Optical spectroscopic diagnosis of cancer and normal breast tissues

J. Opt. Soc. Am. B

Laser induced fluorescence spectroscopy (LIFS): can it be used in the diagnosis and treatment of colonic malignancy?

Am. J. Gastroenterol.

Spectroscopic diagnosis of colonic dysplasia

Photochem. Photobiol.

Multisite topographic microfluorometry of intracellular and exogeneous fluorochromes

Photochem. Photobiol.

Cellular autofluorescence: is it due to flavins?

J. Histochem. Cytochem.