DIY: "Do Imaging Yourself" - Conventional microscopes as powerful tools for in vivo investigation.

Intravital imaging has been increasingly employed in cell biology studies and it is becoming one of the most powerful tools for in vivo investigation. Although some protocols can be extremely complex, most intravital imaging procedures can be performed using basic surgery and animal maintenance techniques. More importantly, regular confocal microscopes - the same that are used for imaging immunofluorescence slides - can also acquire high quality intravital images and movies after minor adaptations. Here we propose minimal adaptations in stock microscopes that allow major improvements in different fields of scientific investigation.

Basics of Confocal Microscopy and the Complexity of Diagnosing Skin Tumors: New Imaging Tools in Clinical Practice, Diagnostic Workflows, Cost-Estimate, and New Trends.

The use of reflectance confocal microscopy (RCM) and other noninvasive imaging devices can potentially streamline clinical care, leading to more precise and efficient management of skin cancer. This article explores the potential role of RCM in cutaneous oncology, as an adjunct to more established techniques of detecting and monitoring for skin cancer, such as dermoscopy and total body photography. Discussed are current barriers to the adoption of RCM, diagnostic workflows and standards of care in the United States and Europe, and medicolegal issues. The potential role of RCM and other similar technological innovations in the enhancement of dermatologic care is evaluated.

[Reflectance confocal microscopy : what future for dermatology ?] / Microscopie confocale in vivo : quel avenir en dermatologie ?

Reflectance confocal microscopy is a non invasive imaging technique which provides in vivo and real time images of different skin tissues with a resolution close to histology, however with a depth limited to superficial dermis.The first lesions that were morphologically analyzed are melanocytic lesions. Reflectance confocal microscopy has been used for about ten years in dermatology. Its progressive improvement over the years has allowed it to be an efficient tool for diagnosing cutaneous tumors. It has been developed for inflammatory dermatosis, cutaneous infections, angiomas, cosmetology. Furthermore, it is also used to delimit the edges of lesions or the area to biopsy. This cutaneous imaging technique represents a major innovation and has its place in dermatological practice.

La microscopie confocale (MC) par réflectance est une technique d'imagerie non invasive qui permet d'obtenir in vivo et en temps réel des images de différents tissus de la peau avec une résolution proche de l'histologie, avec toutefois une profondeur limitée au derme superficiel.Les premières lésions qui ont été analysées morphologiquement sont les lésions mélanocytaires. Utilisée depuis une dizaine d'années en dermatologie, la MC est arrivée à maturité pour le diagnostic des tumeurs cutanées. Elle se développe pour les dermatoses inflammatoires, les infections cutanées, les angiomes, la cosmétologie mais aussi lors d'excision pour préciser les limites d'une lésion ou la zone d'intérêt à biopsier. Cette technique d'imagerie cutanée représente une innovation majeure et a une place logique en pratique dermatologique.

Advances in endoscopic imaging in ulcerative colitis.

Modern strategies for the treatment of ulcerative colitis require more accurate tools for gastrointestinal imaging to better assess mucosal disease activity and long-term prognostic clinical outcomes. Recent advances in gastrointestinal luminal endoscopy are radically changing the role of endoscopy in every-day clinical practice and research trials. Advanced endoscopic imaging techniques including high-definition endoscopes, optical magnification endoscopy, and various chromoendoscopy techniques have remarkably improved endoscopic assessment of ulcerative colitis. More recently, optical biopsy techniques with either endocytoscopy or confocal laser endomicroscopy have shown great potential in predicting several histological changes in real time during ongoing endoscopy. Here, we review current applications of advanced endoscopic imaging techniques in ulcerative colitis and present the most promising upcoming headways in this field.

Recent trends in diagnostic techniques for inflammatory bowel disease.

Although ileocolonoscopy is the gold standard for diagnosis of inflammatory bowel disease and is useful for assessing the disease severity in the colon and terminal ileum, several alternative diagnostic techniques have been developed recently. For ulcerative colitis (UC), magnification colonoscopy, endocytoscopy, and confocal laser endomicroscopy enable assessment of histological inflammation without the need for biopsy. Capsule endoscopy is useful for detection of small intestinal and colonic lesions in both female and male patients. For UC, capsule endoscopy may be useful for evaluating colonic inflammation in patients with a previous poor colonoscopy experience, while it should be used only in Crohn's disease (CD) patients with unexplained symptoms when other examinations are negative. Magnetic resonance enterography (MRE) is particularly useful for detecting transmural inflammation, stenosis, and extraintestinal lesions, including abscesses and fistulas. MRE is also useful when evaluating small and large intestinal lesions, even in cases with severe strictures in which full evaluation of the small bowel would be virtually impossible using other devices. Therefore, the appropriate diagnostic devices for detecting CD lesions in the small and large intestine should be used.

Novel imaging modalities for immune cell monitoring in the intestine.

PURPOSE OF REVIEW: The introduction of novel molecular imaging modalities that can not only define disease states on the basis of structural changes and morphology, but also allow in-vivo visualization and characterization of molecular and biochemical alterations on a cellular level add a new dimension to our current diagnostic possibilities. The advents of innovative endoscopic devices coupled with the introduction of novel targeting ligands contribute to the recent advances made in the field of molecular imaging. The purpose of this review is to present and discuss the concepts and the potential of novel endoscopic imaging modalities for immune cell monitoring in the intestine. RECENT FINDINGS: Recent progress concerning molecular imaging studies in animals and human patients implicates that this approach can be used to improve detection of mucosal lesions in wide-field imaging and for in-vivo characterization of the mucosa with the ultimate goal of assessing the likelihood of response to targeted therapy with biological agents. In particular, molecular endomicroscopy for assessment of mucosal immune responses ('immunoendoscopy') emerges as a novel approach for optimized endoscopic diagnosis and individualized therapy. SUMMARY: Molecular imaging modalities in the intestine have the immediate potential to have an impact on current clinical practice and could therefore open new frontiers for clinical endoscopy and give hope for improved diagnosis and targeted therapies.

Screening, early detection, education, and trends for melanoma: current status (2007-2013) and future directions: Part II. Screening, education, and future directions.

New evidence has accumulated over the past several years that supports improved melanoma outcomes associated with both clinician and patient screening. Population-based and workplace studies conducted in Australia and the Unites States, respectively, have shown decreases in the incidence of thick melanoma and overall melanoma mortality, and a year-long statewide screening program in Germany has shown a nearly 50% reduction in mortality 5 years after the screening ended. Current melanoma screening guidelines in the United States are inconsistent among various organizations, and therefore rates of both physician and patient skin examinations are low. As policymaking organizations update national screening recommendations in the United States, the latest research reviewed in part II of this continuing medical education article should be considered to establish the most effective recommendations. Patient and provider education will be necessary to ensure that appropriate patients receive recommended screening.

Laser scanning confocal endomicroscopy in the neurosurgical operating room: a review and discussion of future applications.

Laser scanning confocal endomicroscopy (LSCE) is an emerging technology for examining brain neoplasms in vivo. While great advances have been made in macroscopic fluorescence in recent years, the ability to perform confocal microscopy in vivo expands the potential of fluorescent tumor labeling, can improve intraoperative tissue diagnosis, and provides real-time guidance for tumor resection intraoperatively. In this review, the authors highlight the technical aspects of confocal endomicroscopy and fluorophores relevant to the neurosurgeon, provide a comprehensive summary of LSCE in animal and human neurosurgical studies to date, and discuss the future directions and potential for LSCE in neurosurgery.

In vivo confocal microscopy in dermatology: from research to clinical application.

Confocal laser scanning microscopy (CLSM) represents an emerging technique for the noninvasive histomorphological analysis of skin in vivo and has shown its applicability for dermatological research as well as its value as an adjunct tool in the clinical management of skin cancer patients. Herein, we aim to give an overview on the current clinical indications for CLSM in dermatology and also highlight the diverse applications of CLSM in dermatological research.

Optical endomicroscopy and the road to real-time, in vivo pathology: present and future.

Epithelial cancers account for substantial mortality and are an important public health concern. With the need for earlier detection and treatment of these malignancies, the ability to accurately detect precancerous lesions has an increasingly important role in controlling cancer incidence and mortality. New optical technologies are capable of identifying early pathology in tissues or organs in which cancer is known to develop through stages of dysplasia, including the esophagus, colon, pancreas, liver, bladder, and cervix. These diagnostic imaging advances, together as a field known as optical endomicroscopy, are based on confocal microscopy, spectroscopy-based imaging, and optical coherence tomography (OCT), and function as "optical biopsies," enabling tissue pathology to be imaged in situ and in real time without the need to excise and process specimens as in conventional biopsy and histopathology. Optical biopsy techniques can acquire high-resolution, cross-sectional images of tissue structure on the micron scale through the use of endoscopes, catheters, laparoscopes, and needles. Since the inception of these technologies, dramatic technological advances in accuracy, speed, and functionality have been realized. The current paradigm of optical biopsy, or single-area, point-based images, is slowly shifting to more comprehensive microscopy of larger tracts of mucosa. With the development of Fourier-domain OCT, also known as optical frequency domain imaging or, more recently, volumetric laser endomicroscopy, comprehensive surveillance of the entire distal esophagus is now achievable at speeds that were not possible with conventional OCT technologies. Optical diagnostic technologies are emerging as clinically useful tools with the potential to set a new standard for real-time diagnosis. New imaging techniques enable visualization of high-resolution, cross-sectional images and offer the opportunity to guide biopsy, allowing maximal diagnostic yields and appropriate staging without the limitations and risks inherent with current random biopsy protocols. However, the ability of these techniques to achieve widespread adoption in clinical practice depends on future research designed to improve accuracy and allow real-time data transmission and storage, thereby linking pathology to the treating physician. These imaging advances are expected to eventually offer a see-and-treat paradigm, leading to improved patient care and potential cost reduction. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/5372548637202968.

Endoscopic diagnosis of biliary tract disease.

PURPOSE OF REVIEW: Endoscopic diagnosis of biliary disease is challenging due to difficulties in access, visualization, and sampling. Recent advances in endoscopic technology, ancillary diagnostic methods, and our understanding of autoimmune pancreatitis (AIP) and IgG4-related cholangitis (IRC) have led to improvements in the endoscopic diagnosis of pancreaticobiliary disease. RECENT FINDINGS: Single-operator cholangioscopy overcomes several of the limitations of mother-baby cholangioscopy enhancing the diagnostic accuracy in indeterminate pancreaticobiliary disease. Probe-based confocal laser endomicroscopy has been recently shown to provide a significantly higher accuracy for the diagnosis of malignant biliary strictures than achieved by endoscopic retrograde cholangiopancreatogram and standard tissue acquisition, and has the potential to develop into a useful adjunct method of cholangioscopy. Fluorescence in-situ hybridization increases the sensitivity of routine brush cytology without compromising specificity in patients with indeterminate biliary strictures. The diagnosis of AIP/IRC remains challenging. The recently published international consensus criteria for AIP have included data on the potential diagnostic utility of endoscopic retrograde pancreatogram and endoscopic ampullary biopsies. SUMMARY: Recent technical advances as well as ancillary diagnostic methods have improved the diagnostic accuracy of conventional endoscopic techniques. Future refinement of endoscopic methods may further improve diagnostic approaches to biliary disease.

Confocal laser endomicroscopy: a primer for pathologists.

CONTEXT: The advent of new endoscopic optical techniques is likely to change pathologists' role in diagnosis. OBJECTIVE: To describe how confocal laser endomicroscopy (CLE) works, show its advantages and limitations compared to cytohistologic biopsy, and explore how it may affect the practice of pathology. DATA SOURCES: Literature review. CONCLUSIONS: Confocal laser endomicroscopy is proving its ability to provide histology-like images of tissues in vivo to help avoid risks and costs of conventional biopsies. Confocal imaging restricts light to 1 plane, emulating a paraffin section, and topical or systemic optical contrast agents allow subcellular resolution. New contrast agents could theoretically permit molecular characterization. In vivo imaging has begun to demonstrate novel, dynamic types of diagnostic features. Decreased histologic biopsies can be anticipated for a few scenarios. Significant limitations of CLE include the inability to create a tissue archive for broad molecular classification, suboptimal contrast agents, small fields of view and shallow penetration, paucity of clinical validation studies, and problems with reimbursement. Confocal laser endomicroscopy exposes new opportunities for pathologists: CLE technologies can be exploited in pathology, and diagnostic criteria expanded based on endoscopists' discoveries. Potential synergy exists between CLE and cytology, allowing the low-magnification diagnostic architectural changes by CLE and cytomorphology to emulate the full diagnostic information in a histologic biopsy while providing an archive of material for molecular or immunohistochemical studies. Confocal laser endomicroscopy will decrease some types of biopsies, but offers an opportunity for pathologists to find new ways to provide value and improve patient care.

New imaging techniques and opportunities in endoscopy.

Gastrointestinal endoscopy is undergoing major improvements, which are driven by new available technologies and substantial refinements of optical features. In this Review, we summarize available and evolving imaging technologies that could influence the clinical algorithm of endoscopic diagnosis. Detection, characterization and confirmation are essential steps required for proper endoscopic diagnosis. Optical and nonoptical methods can help to improve each step; these improvements are likely to increase the detection rate of neoplasias and reduce unnecessary endoscopic treatments. Furthermore, functional and molecular imaging are emerging as new diagnostic tools that could provide an opportunity for personalized medicine, in which endoscopy will define disease outcome or predict the response to targeted therapy.

Progress toward optical biopsy: bringing the microscope to the patient.

The investigation of many lung diseases currently requires bronchoscopic or surgical histopathological tissue biopsy. This creates risks for patients and entails processing costs and delays in diagnosis. However, several mainly probe-based biophotonic techniques that can image solitary lesions and diffuse lung diseases are fuelling a paradigm shift toward real-time in vivo diagnosis. Optical coherence tomography (OCT) uses near-infrared light in a process analogous to ultrasonography to image the mucosal and submucosal tissue boundaries of the bronchial tree. With 15-µm resolution, early work suggests it can differentiate between neoplasia, carcinoma in situ, dysplasia, and metaplasia based around epithelial thickness and breaches in the basement membrane. Probe-based confocal laser endomicroscopy (pCLE) has superior resolution but less penetration than OCT and employs blue argon laser light to fluoresce the endogenous elastin of (1) the acinar scaffold of the peripheral lung and (2) the basement membrane lying under bronchial mucosa. Initial studies suggest that the regular fibre arrangement of the basement membrane is altered in the presence of overlying malignant epithelium. pCLE produces detailed representations of the alveolar septal walls, microvessels, and some inflammatory cells. A third device, the endocytoscope, is a contact microscope requiring contrast agent to provide subcellular resolution of bronchial mucosa. Further development of these "optical biopsy" techniques and evaluation of diagnostic sensitivity and specificity of the acquired images are needed before they can be considered effective methods for eliminating the need for, and thus risks of, pinch biopsy to enable real-time diagnosis to streamline management.

In vivo confocal microscopy, an inner vision of the cornea - a major review.

The demands of modern ophthalmology have evolved from descriptive findings from the slit lamp to in vivo assessment of cellular level changes. Nowadays, the latter can be provided by in vivo confocal microscopy. This article gives an overview of confocal principles using tandem scanning, scanning slit and laser scanning techniques used in ophthalmology. The main part of the paper describes the clinical applications emphasizing the anatomy of the normal and pathological cornea, and illustrates side-effects of topical medication, contact lens wear, cross-linking and refractive surgery. Finally, a summary about experimental applications, including animal studies, surface characterization and volume rendering as well as future developments, is given.

Chromoscopic endomicroscopy: in vivo cellular resolution imaging of the colorectum.

Advances in imaging technology and engineering have now permitted functional integration of a confocal endomicroscope into the distal tip of a conventional video colonoscope enabling imaging of the surface epithelium and the underlying lamina propria during ongoing video endoscopy. For the first time, the endoscopist is now able to resolve the surface and subsurface mucosa at cellular resolution in vivo and in real time. A new era in endoscopic imaging has therefore begun - histoendoscopy. In addition to providing a high-accuracy in vivo optical biopsy tool for the differentiation between benign hyperplasia, intra-epithelial neoplasia and carcinoma in sporadic cohorts, endomicroscopy with targeted biopsies has now been shown to increase the yield of intra-epithelial neoplasia complicating ulcerative colitis. Furthermore, recent data examining endomicroscopic molecular ex vivo imaging using anti-CD44v6 antibody has identified aberrant crypt foci based on their surface molecular expression. Receptor overexpression in vivo in humans may, in the near future, be exploited for the diagnosis of inflammation, neoplasia and in predicting targeted molecular therapy. Endomicroscopy will be key to this immuno-imaging interface. Within the present review, we discuss the current clinical evidence in support of confocal endomicroscopy and explore the new diagnostic possibilities for this technology.