Fibre-based fluorescence-lifetime imaging microscopy: a real-time biopsy guidance tool for suspected lung cancer.

Lung cancer is the most common cause of cancer-related deaths worldwide. Early detection improves outcomes, however, existing sampling techniques are associated with suboptimal diagnostic yield and procedure-related complications. Autofluorescence-based fluorescence-lifetime imaging microscopy (FLIM), a technique which measures endogenous fluorophore decay rates, may aid identification of optimal biopsy sites in suspected lung cancer. Our fibre-based fluorescence-lifetime imaging system, utilising 488 nm excitation, which is deliverable via existing diagnostic platforms, enables real-time visualisation and lifetime analysis of distal alveolar lung structure. We evaluated the diagnostic accuracy of the fibre-based fluorescence-lifetime imaging system to detect changes in fluorescence lifetime in freshly resected ex vivo lung cancer and adjacent healthy tissue as a first step towards future translation. The study compares paired non-small cell lung cancer (NSCLC) and non-cancerous tissues with gold standard diagnostic pathology to assess the performance of the technique. Paired NSCLC and non-cancerous lung tissues were obtained from thoracic resection patients (N=21). A clinically compatible 488 nm fluorescence-lifetime endomicroscopy platform was used to acquire simultaneous fluorescence intensity and lifetime images. Fluorescence lifetimes were calculated using a computationally-lightweight, rapid lifetime determination method. Fluorescence lifetime was significantly reduced in ex vivo lung cancer, compared with non-cancerous lung tissue [mean ± standard deviation (SD), 1.79±0.40 vs. 2.15±0.26 ns, P<0.0001], and fluorescence intensity images demonstrated distortion of alveolar elastin autofluorescence structure. Fibre-based fluorescence-lifetime imaging demonstrated good performance characteristics for distinguishing lung cancer, from adjacent non-cancerous tissue, with 81.0% sensitivity and 71.4% specificity. Our novel fibre-based fluorescence-lifetime imaging system, which enables label-free imaging and quantitative lifetime analysis, discriminates ex vivo lung cancer from adjacent healthy tissue. This minimally invasive technique has potential to be translated as a real-time biopsy guidance tool, capable of optimising diagnostic accuracy in lung cancer.

Full spectrum fluorescence lifetime imaging with 0.5 nm spectral and 50 ps temporal resolution.

The use of optical techniques to interrogate wide ranging samples from semiconductors to biological tissue for rapid analysis and diagnostics has gained wide adoption over the past decades. The desire to collect ever more spatially, spectrally and temporally detailed optical signatures for sample characterization has specifically driven a sharp rise in new optical microscopy technologies. Here we present a high-speed optical scanning microscope capable of capturing time resolved images across 512 spectral and 32 time channels in a single acquisition with the potential for ~0.2 frames per second (256 × 256 image pixels). Each pixel in the resulting images contains a detailed data cube for the study of diverse time resolved light driven phenomena. This is enabled by integration of system control electronics and on-chip processing which overcomes the challenges presented by high data volume and low imaging speed, often bottlenecks in previous systems.

Solitary pulmonary nodule imaging approaches and the role of optical fibre-based technologies.

Solitary pulmonary nodules (SPNs) are a clinical challenge, given there is no single clinical sign or radiological feature that definitively identifies a benign from a malignant SPN. The early detection of lung cancer has a huge impact on survival outcome. Consequently, there is great interest in the prompt diagnosis, and treatment of malignant SPNs. Current diagnostic pathways involve endobronchial/transthoracic tissue biopsies or radiological surveillance, which can be associated with suboptimal diagnostic yield, healthcare costs and patient anxiety. Cutting-edge technologies are needed to disrupt and improve, existing care pathways. Optical fibre-based techniques, which can be delivered via the working channel of a bronchoscope or via transthoracic needle, may deliver advanced diagnostic capabilities in patients with SPNs. Optical endomicroscopy, an autofluorescence-based imaging technique, demonstrates abnormal alveolar structure in SPNs in vivo Alternative optical fingerprinting approaches, such as time-resolved fluorescence spectroscopy and fluorescence-lifetime imaging microscopy, have shown promise in discriminating lung cancer from surrounding healthy tissue. Whilst fibre-based Raman spectroscopy has enabled real-time characterisation of SPNs in vivo Fibre-based technologies have the potential to enable in situ characterisation and real-time microscopic imaging of SPNs, which could aid immediate treatment decisions in patients with SPNs. This review discusses advances in current imaging modalities for evaluating SPNs, including computed tomography (CT) and positron emission tomography-CT. It explores the emergence of optical fibre-based technologies, and discusses their potential role in patients with SPNs and suspected lung cancer.

Deep Learning in ex-vivo Lung Cancer Discrimination using Fluorescence Lifetime Endomicroscopic Images.

Fluorescence lifetime is effective in discriminating cancerous tissue from normal tissue, but conventional discrimination methods are primarily based on statistical approaches in collaboration with prior knowledge. This paper investigates the application of deep convolutional neural networks (CNNs) for automatic differentiation of ex-vivo human lung cancer via fluorescence lifetime imaging. Around 70,000 fluorescence images from ex-vivo lung tissue of 14 patients were collected by a custom fibre-based fluorescence lifetime imaging endomicroscope. Five state-of-the-art CNN models, namely ResNet, ResNeXt, Inception, Xception, and DenseNet, were trained and tested to derive quantitative results using accuracy, precision, recall, and the area under receiver operating characteristic curve (AUC) as the metrics. The CNNs were firstly evaluated on lifetime images. Since fluorescence lifetime is independent of intensity, further experiments were conducted by stacking intensity and lifetime images together as the input to the CNNs. As the original CNNs were implemented for RGB images, two strategies were applied. One was retaining the CNNs by putting intensity and lifetime images in two different channels and leaving the remaining channel blank. The other was adapting the CNNs for two-channel input. Quantitative results demonstrate that the selected CNNs are considerably superior to conventional machine learning algorithms. Combining intensity and lifetime images introduces noticeable performance gain compared with using lifetime images alone. In addition, the CNNs with intensity-lifetime RGB image is comparable to the modified two-channel CNNs with intensity-lifetime two-channel input for accuracy and AUC, but significantly better for precision and recall.

The relationship of nightmare frequency and nightmare distress to well-being.

Nightmares can be defined as very disturbing dreams, the events or emotions of which cause the dreamer to wake up. In contrast, unpleasant dreams can be defined in terms of a negative emotional rating of a dream, irrespective of whether or not the emotions or events of the dream woke the dreamer. This study addresses whether frequency of unpleasant dreams is a better index of low well-being than is frequency of nightmares. A total of 147 participants reported their nightmare frequency retrospectively and then kept a log of all dreams, including nightmares, for 2 weeks, and rated each dream for pleasantness/unpleasantness. Anxiety, depression, neuroticism, and acute stress were found to be associated with nightmare distress (ND) (the trait-like general level of distress in waking-life caused by having nightmares) and prospective frequency of unpleasant dreams, and less so with the mean emotional tone of all dreams, or retrospective or prospective nightmare frequency. Correlations between low well-being and retrospective nightmare frequency became insignificant when trait ND was controlled for, but correlations with prospective unpleasant dream frequency were maintained. The reporting of nightmares may thus be confounded and modulated by trait ND: such confounding does not occur for the reporting of unpleasant dreams in general. Thus there may be attributional components to deciding that one has been awoken by a dream, which can affect estimated nightmare frequency and its relationship with well-being. Underestimation of nightmare frequency by the retrospective questionnaire compared with logs was found to be a function of mean dream unpleasantness and ND.