Performance estimation of diagnostic tests for cervical precancer based on fluorescence spectroscopy: effects of tissue type, sample size, population, and signal-to-noise ratio

Fluorescence spectroscopy may provide a cost-effective tool to improve precancer detection. We describe a method to estimate the diagnostic performance of classifiers based on optical spectra, and to explore the sensitivity of these estimations to factors affecting spectrometer cost. Fluorescence spectra were obtained at three excitation wavelengths in 92 patients with an abnormal Papanicolaou smear and 51 patients with no history of an abnormal smear. Bayesian classification rules were developed and evaluated at multiple misclassification costs. We explored the sensitivity of classifier performance to variations in tissue type, sample size, tested population, signal to noise ratio (SNR), and number of excitation and emission wavelengths. Sensitivity and specificity could be evaluated within +/- 7%. Minimal decrease in diagnostic performance is observed as SNR is reduced to 15, the number of excitation-emission wavelength combinations is reduced to 15 or the number of excitation wavelengths is reduced to one. Diagnostic performance is compromised when ultraviolet excitation is not included. Significant spectrometer cost reduction is possible without compromising diagnostic ability. Decision-analytic methods can be used to rate designs based on incremental cost-effectiveness.     

Ensembles of radial basis function networks for spectroscopicdetection of cervical precancer

The mortality related to cervical cancer can be substantially reduced through early detection and treatment. However, current detection techniques, such as Pap smear and colposcopy, fail to achieve a concurrently high sensitivity and specificity. In vivo fluorescence spectroscopy is a technique which quickly, noninvasively and quantitatively probes the biochemical and morphological changes that occur in precancerous tissue. A multivariate statistical algorithm was used to extract clinically useful information from tissue spectra acquired from 361 cervical sites from 95 patients at 337-, 380-, and 460-nm excitation wavelengths. The multivariate statistical analysis was also employed to reduce the number of fluorescence excitation-emission wavelength pairs required to discriminate healthy tissue samples from precancerous tissue samples. The use of connectionist methods such as multilayered perceptrons, radial basis function (RBF) networks, and ensembles of such networks was investigated. RBF ensemble algorithms based on fluorescence spectra potentially provide automated and near real-time implementation of precancer detection in the hands of nonexperts. The results are more reliable, direct, and accurate than those achieved by either human experts or multivariate statistical algorithms     

Electrical impedance spectroscopy (EIS) in the urinary bladder: the effect of inflammation and edema on identification of malignancy

Previous studies have shown that tetrapolar electrical impedance spectroscopy measurements can identify cervical interstitial neoplasia with the same sensitivity and specificity as cervical smears. In the urinary bladder, the same technique yields significant differences (p < 0.05 at seven frequencies between 9.6 and 614 kHz) between normal and malignant urothelium, but is unable to classify individual measurements. Detailed histological examination demonstrates that inflammation and edema--both of which are common in abnormal urothelium--alter the impedance spectrum significantly in opposing directions. Consideration of morphological changes in abnormal urothelium suggests alternative measurement strategies.     

Comparison of multiexcitation fluorescence and diffuse reflectance spectroscopy for the diagnosis of breast cancer (March 2003)

Nonmalignant (n = 36) and malignant (n = 20) tissue samples were obtained from breast cancer and breast reduction surgeries. These tissues were characterized using multiple excitation wavelength fluorescence spectroscopy and diffuse reflectance spectroscopy in the ultraviolet-visible wavelength range, immediately after excision. Spectra were then analyzed using principal component analysis (PCA) as a data reduction technique. PCA was performed on each fluorescence spectrum, as well as on the diffuse reflectance spectrum individually, to establish a set of principal components for each spectrum. A Wilcoxon rank-sum test was used to determine which principal components show statistically significant differences between malignant and nonmalignant tissues. Finally, a support vector machine (SVM) algorithm was utilized to classify the samples based on the diagnostically useful principal components. Cross-validation of this nonparametric algorithm was carried out to determine its classification accuracy in an unbiased manner. Multiexcitation fluorescence spectroscopy was successful in discriminating malignant and nonmalignant tissues, with a sensitivity and specificity of 70% and 92%, respectively. The sensitivity (30%) and specificity (78%) of diffuse reflectance spectroscopy alone was significantly lower. Combining fluorescence and diffuse reflectance spectra did not improve the classification accuracy of an algorithm based on fluorescence spectra alone. The fluorescence excitation-emission wavelengths identified as being diagnostic from the PCA-SVM algorithm suggest that the important fluorophores for breast cancer diagnosis are most likely tryptophan, NAD(P)H and flavoproteins.     

Immunomorphological identification of an antigen associated with squamous cell carcinoma of the cervix uteri

Data on immunomorphological identification of antigen associated with cervical squamous cell carcinoma are presented. Tissue specimens prepared of cervical squamous cell carcinoma, normal cervical tissue and different other human tumours have been stained immunohistochemically for antigen using a polyclonal antiserum. Immunofluorescence staining for tumour-associated antigen was observed in all specimens of cervical squamous cell carcinoma and in 20% of specimens of the normal cervix uteri. The above antigen is a high specific tumour-associated marker and may be useful in the diagnostic studies.     

LED激发光源的水体浮游植物浓度活体检测系统研究

采用5种不同中心激发波长的超高亮LED作为激发光源构建了一套水体浮游植物活体荧光检测系统.利用该系统测量24种不同来源水样和同一水样10种不同比例稀释样品的浮游植物活体荧光强度,研究了荧光强度与叶绿素a浓度的线性关系.结果表明:蓝光LED(最大发射波长442.7 nm)是用以测量浮游植物活体荧光的最适合光源,相应的检测限达到了0.0278 μg/l(叶绿素a);浮游植物种类、环境造成的荧光量子效率以及水体DOM背景荧光均对活体荧光的测量结果产生影响,尤以浮游植物种类和荧光量子效率的影响更为显著.因此,蓝光超高亮LED将为浮游植物活体荧光测量提供廉价的替代光源,为利用该光源研发浮游植物浓度原位荧光检测仪提供了潜在的可能.为克服浮游植物种类、荧光量子效率以及DOM背景荧光对浮游植物活体荧光测量结果的影响,需要进一步研究上述因素的具体影响行为,发展相应方法进行校正.     

光学相干层析术及其应用

光学相干层析术是一种新型的对活体组织进行非侵入的光学诊断成像技术。它将低相干干涉仪与共焦扫描显微术结合在一起，去掉物镜焦点之外的散射光，利用高灵敏度的探测技术，实现在光散射介质如生物组织中获得清晰的分层图像。     

Real-time sleep apnea detection by classifier combination

To find an efficient and valid alternative of polysomnography (PSG), this paper investigates real-time sleep apnea and hypopnea syndrome (SAHS) detection based on electrocardiograph (ECG) and saturation of peripheral oxygen (SpO(2)) signals, individually and in combination. We include ten machine-learning algorithms in our classification experiment. It is shown that our proposed SpO (2) features outperform the ECG features in terms of diagnostic ability. More importantly, we propose classifier combination to further enhance the classification performance by harnessing the complementary information provided by individual classifiers. With our selected SpO(2) and ECG features, the classifier combination using AdaBoost with Decision Stump, Bagging with REPTree, and either kNN or Decision Table achieves sensitivity, specificity, and accuracy all around 82% for a minute-based real-time SAHS detection over 25 sleep-disordered-breathing suspects' full overnight recordings.     

Hyperspectral Fluorescence Imaging for Mouse Skin Tumor Detection

This paper presents a hyperspectral imaging technique based on laser‐induced fluorescence for non‐invasive detection of tumorous tissue on mouse skin. Hyperspectral imaging sensors collect image data in a number of narrow, adjacent spectral bands. Such high‐resolution measurement of spectral information reveals contiguous emission spectra at each image pixel useful for the characterization of constituent materials. The hyperspectral image data used in this study are fluorescence images of mouse skin consisting of 21 spectral bands in the visible spectrum of the wavelengths ranging from 440 nm to 640 nm. Fluorescence signal is measured with the use of laser excitation at 337 nm. An acousto‐optic tunable filter (AOTF) is used to capture images at 10 nm intervals. All spectral band images are spatially registered with the reference band image at 490 nm to obtain exact pixel correspondences by compensating the spatial offsets caused by the refraction differences in AOTF at different wavelengths during the image capture procedure. The unique fluorescence spectral signatures demonstrate a good separation to differentiate malignant tumors from normal tissues for rapid detection of skin cancers without biopsy.     

几种人工合成色素的荧光光谱与拉曼光谱研究

应用荧光光谱仪及微拉曼光谱仪分别对几种常见的人工合成色素溶液的荧光光谱和拉曼光谱进行了实验研究。一方面,首次测量其不同浓度合成色素溶液的荧光光谱,讨论其谱线特性。另一方面,以有致癌性的合成色素苋菜红为例,应用拉曼光谱技术进行定性和定量检测,验证拉曼光谱对色素检测的可行性。为了提高探测灵敏度,以55nm的金纳米粒子溶胶为基底,在785nm激发光下对不同浓度的苋菜红溶液进行了表面增强拉曼光谱探测,探测到的最低浓度为10-17mol/L。分析结果为这种色素的合理利用及安全快速检测提供了有效的光谱实验依据,也为其它色素的检测提供了参考方法。     

高斯分解法研究浮游植物荧光激发光谱

研究了甲藻和硅藻两个门类的六种浮游植物在发射波长为675 nm处的活体叶绿素荧光激发光谱.对其作四阶导数分析的基础上,根据导数光谱中出现的极大值位置和数量,对甲藻门和硅藻门的激发光谱分别设定统一的初始中心波长.以初始波长为中心对激发光谱做高斯分解及多峰拟合,建立门类水平上统一的高斯基库.研究发现,在波长为350~550 nm内,甲藻和硅藻的高斯基库特征差异显著,而同门类的高斯基库特征相似,表明高斯分解法为甲藻和硅藻的分类识别提供有效的技术手段.利用高斯基库的参数拟合原光谱,拟合曲线与原光谱能较好地吻合,误差小.高斯分解法可再现活体激发光谱中重叠的色素荧光峰,也为光合色素的活体测定提供了新的方法.     

Detection of rapid-eye movements in sleep studies

One of the key features of rapid-eye movement (REM) sleep is the presence of bursts of REMs. Sleep studies routinely use REMs to classify sleep stages. Moreover, REM count or density has been used in studies involving learning and various psychiatric disorders. Most of these studies have been based on the visual identification of REMs, which is generally a very time-consuming task. This and the varying definitions of REMs across scorers have warranted the development of automatic REM detection methodologies. In this paper, we present a new detection scheme that combines many of the intrinsic properties of REMs and requires minimal parameter adjustments. In the proposed method, a single parameter can be used to control the REM detection sensitivity and specificity tradeoff. Manually scored training data are used to develop the method. We assess the performance of the method against manual scoring of individual REM events and present validation results using a separate data set. The ability of the method to discriminate fast horizontal ocular movement in REM sleep from other types of events is highlighted. A key advantage of the presented method is the minimal a priori information requirement. The results of training data (recordings from five subjects) show an overall sensitivity of 78.8% and specificity of 81.6%. The performance on the testing data (recording from five subjects different from the training data) showed overall sensitivity of 67.2% and specificity of 77.5%.     

A novel optical imaging method for the early detection, quantitative grading, and mapping of cancerous and precancerous lesions of cervix

This paper describes a novel optical imaging method for the in vivo early detection, quantitative staging, and mapping of cervical cancer and precancer. A multispectral imaging system was developed, which is capable of performing time-resolved imaging spectroscopy. The system was used in order to assess quantitatively the alterations in the light scattering properties of the cervix, induced selectively and reversibly in cervical neoplasias, after the application of acetic acid solution. Spectral imaging and analysis of cervix show that the maximum contrast between acetic acid responsive and nonresponsive areas is obtained at 525 +/- 15 nm, which is further enhanced by cutting off the regular component of tissue reflection, with the aid of two linear cross polarizers. Successive snapshot imaging at this spectral band enables the quantitative assessment of the temporal alterations in the intensity of the backscattered light, in any spatial location of the examined area. Initial clinical trials show that optical contrast enhancement results in a notable improvement of the sensitivity in detecting incipient lesions. It was also shown that the measured temporal characteristics of the phenomenon contain specific information, which enables the differentiation between neoplastic and nonneoplastic lesions, as well as between neoplasias of different grade. The demonstrated improved sensitivity and specificity highlight the potential of the method in both clinical research and noninvasive diagnosis.     

Using a rapid communication approach to improve a POC Chlamydia test

Chlamydia trachomatis is a common sexually transmitted infection in young women. Available point-of-care (POC) diagnostic tests perform poorly, but development of new devices can be costly and time consuming. We explored the feasibility (user friendliness) and test characteristics (sensitivity and specificity) of a new prototype device to detect Chlamydia in adolescent women by using small numbers of subjects and rapid communication with the manufacturer. We compared cervical POC test results to the gold standard (cervical nucleic acid amplification testing). We also assessed the accuracy of the POC test on self-collected vaginal swabs by comparing results to cervical nucleic acid amplification test and to the cervical POC test. We frequently reviewed user experience and test results with the manufacturer. The results showed the feasibility and accuracy of the device. Feasibility--initial device malfunctions were identified and corrected. This device would be easy to use in a nonclinical setting, as it is self-contained and the color change for some specimens was dramatic and immediate. Accuracy--initial prototypes demonstrated low sensitivities (38%) for vaginal and cervical swabs. After feedback, the company developed new prototypes with improved sensitivity (80%). However, the increased sensitivity was accompanied by a high percentage of indeterminate results and false positives that lowered specificity.     

High Performance,Multiplexed Lung Cancer Biomarker Detection on a Plasmonic Gold Chip

Diagnosis of lung cancer is performed using a plasmonic gold (pGOLD) chip through multiplexed near‐infrared (NIR) detection of carcino‐embryonic antigen (CEA), Cyfra21‐1, and neuron‐specific enolase (NSE) in the serum samples of patients. With ≈50‐fold enhancement of NIR fluorescence, multiplexed microarray analysis of CEA, Cyfra21‐1, and NSE in 10 μL of human serum or whole blood samples on pGOLD chip leads to markedly improved limit‐of‐quantification, limit‐of‐detection, reproducibility, and higher diagnostic sensitivity and specificity compared to traditional biochips and Luminex technology currently in use in hospitals.     

饱和激发情况下油荧光非线性特征初步分析

利用激光诱导荧光( LIF)技术,从荧光强度和光谱偏移两个角度研究饱和激发状态下油荧光的非线性变化特征。利用能量变化的激光束诱导油样本荧光,获得荧光强度变化数据,通过非线性曲线拟合来标定荧光特征峰位430nm处的强度变化参数,获得该波长荧光强度的线性变化范围和非线性趋势,并利用探测波长处荧光强度构造出随诱导光强变化的三维油荧光光谱,实现基于激发光能量参量的油样本光谱区分。实验中,针对普通荧光光谱法无法区分的油样本,结合饱和激发状态下油荧光非线性变化和基于激发光能量参量的油样本光谱特征,实现了样本的有效区分。     

A highly sensitive rapid diagnostic test for Chagas disease that utilizes a recombinant Trypanosoma cruzi antigen

Improved diagnostic tests for Chagas disease are urgently needed. A new lateral flow rapid test for Chagas disease is under development at PATH, in collaboration with Laboratorio Lemos of Argentina, which utilizes a recombinant antigen for detection of antibodies to Trypanosoma cruzi. To evaluate the performance of this test, 375 earlier characterized serum specimens from a region where Chagas is endemic were tested using a reference test (the Ortho T. cruzi ELISA, Johnson & Johnson), a commercially available rapid test (Chagas STAT-PAK, Chembio), and the PATH-Lemos rapid test. Compared to the composite reference tests, the PATH-Lemos rapid test demonstrated an optimal sensitivity of 99.5% and specificity of 96.8%, while the Chagas STAT-PAK demonstrated a sensitivity of 95.3% and specificity of 99.5%. These results indicate that the PATH-Lemos rapid test shows promise as an improved and reliable tool for screening and diagnosis of Chagas disease.     

Laser-induced time-resolved fluorescence of vegetation

Time-resolved measurements of the laser-induced chlorophyll fluorescence emission of vegetation detected by two different techniques are described. Fluorescence decay time measurements using single photon counting and picosecond laser pulse excitation have been used to analyze the fluorescence heterogeneity of plant leaves. The fluorescence is described by lifetimes of 10-40, 80-150, 400-500, and 700-1000 ps. By closing the reaction centers via application of the herbicide DCMU, the lifetimes of the two slowest components increase by a factor of about three. Another possible method to monitor the fluorescence after picosecond excitation could be a streak camera detection system. Measurements performed on the slow decay component of stressed and unstressed plants are presented     

Upconversion Luminescent Chemodosimeter Based on NIR Organic Dye for Monitoring Methylmercury In Vivo

While most luminescent organic dyes display intense Stokes fluorescence, some of them exhibit unique single‐photon frequency upconversion luminescence (FUCL). Compared to conventional anti‐Stokes luminescence of lanthanides and two‐photon excitation, FUCL materials display adjustable spectrum area and require a much lower excitation power. Although this is very beneficial for biological applications in the perspective of reducing photodamage to biological samples and photobleaching of the dyes, the utilization of FUCL for biosensing and bioimaging in vivo has not been reported. In this study, we developed a near‐infrared (NIR) rhodamine derivative (FUC‐1) as a chemodosimeter, which displays weak luminescence but undergoes thiolactone ring‐open process leading to luminescence turn‐on in response to mercury(II) cation or methylmercury with good selectivity and high sensitivity in aqueous solution. Interestingly, FUC‐1 displays particular FUCL, excitation at 808 nm leads to luminescence at 745 nm. Compared to Stokes luminescence resulted from excitation at 630 nm, the use of FUCL lowers the detection limit of Hg²⁺ to be 0.207 nM. FUC‐1 has been used for FUCL bioimaging of methylmercury in live cells and mice. To the best of our knowledge, this is the first example of FUCL biosensing and bioimaging in vivo using visible and NIR fluorescence of small‐molecular dyes.