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Confocal Microscopy: Clinical Impacts and Innovation

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A special issue of Diagnostics (ISSN 2075-4418). This special issue belongs to the section "Optical Diagnostics".

Deadline for manuscript submissions: closed (31 August 2023) | Viewed by 24357

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Prof. Dr. Cleopatra Charalampaki

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Guest Editor

Department of Neurosurgery, Cologne Medical Center, University Witten, Cologne, Germany
Interests: optic; photonic; confocal microscopy; modern operative techniques; deep machine learning; molecular pathology
Special Issues, Collections and Topics in MDPI journals

Dr. Stamatia Giannarou

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Guest Editor

Department of Electrical and Electronic Engineering, Imperial College London, London, UK
Interests: enhanced surgical vision for navigation during minimally invasive and robot-assisted operations

Special Issue Information

Dear Colleagues,

Tumor diagnosis and definition of tumor borders intraoperatively are based on visualization modalities used by surgeons as well as on the histopathologic examination of a limited number of biopsy specimens. Furthermore, optimal surgical therapy is the combination of maximal near-total resection and minimal injury of normal tissues, achieved only if it is possible to identify intraoperative cellular structures and differentiate tumor tissue from normal functional tissue, so to resect a tumor completely and protect the normal tissue. To achieve this goal, we need new technological equipment combined with new surgical concepts.

The principle of confocal laser endomicroscopy (CLE), based on extreme miniaturization of the microscope imaging head, offers the possibility of in vivo microscopy with subcellular and subnuclear resolution. Clear visualization of the cytoarchitecture can be achieved with a 400-fold to 1000-fold magnification.

In oncological diagnosis and surgery, CLE would allow on one hand, intraoperative detection and differentiation of single tumor cells (without the need of fast biopsies) and on the other hand, the definition of borders between tumor and normal tissue on a cellular level, making surgical resection much more accurate than ever before. The application and implementation of CLE-assisted surgery in surgical oncology increases not only the diagnostic but also the therapeutic options by extending the resection borders of cancer on a cellular level and, more importantly, by protecting the functionality of normal tissues.

For this Special Issue, we would like to attract studies from authors working on implementing in vivo and ex vivo surgical and clinical concepts and techniques of confocal laser endomicroscopy in their treatments.

Potential topics of interest include but are not limited to the following:

Confocal-assisted surgery for oncologic brain and head and neck surgery as well as for other disciplines related to oncological surgery
New frontiers in fluorescence microscopy used for surgical oncology
Advances in intraoperative visualization modalities regarding tumor differentiation
Implementation of cell imaging in different oncologic working disciplines
New concept of cellular surgical cancer therapies in the operation theatre
Embedded technological systems for tumor margins differentiation and cellular analysis in thoracic, abdominal, and urological diseases
Deep machine learning techniques for confocal microscopy in pathology

Prof. Dr. Cleopatra Charalampaki
Dr. Stamatia Giannarou
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Diagnostics is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

confocal laser endomicroscopy
diagnosis
biopsy specimens
tumor
cell imaging
deep machine learning techniques

Related Special Issue

Confocal Microscopy: Clinical Impacts and Innovation—2nd Edition in Diagnostics

Published Papers (10 papers)

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Editorial

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3 pages, 175 KiB

Open AccessEditorial

Confocal Laser Endomicroscopy in Oncological Surgery

by Patra Charalampaki and Irini Kakaletri

Diagnostics 2021, 11(10), 1813; https://doi.org/10.3390/diagnostics11101813 - 30 Sep 2021

Cited by 2 | Viewed by 1366

Abstract

The therapy of choice in the treatment of abnormalities in the human body, is to attempt a personalized diagnosis and with minimal invasiveness, ideally resulting in total resection (surgery) or turning off (intervention) of the pathology with preservation of normal functional tissue, followed [...] Read more.

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

Research

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12 pages, 3211 KiB

Open AccessCommunication

Vitiligo Treated with Combined Piperine-Based Topical Treatment and Narrowband Ultraviolet B Therapy: Follow-Up with Reflectance Confocal Microscopy

by Cristina Bertoli, Johanna Chester, Chiara Cortelazzi, Silvana Ciardo, Marco Manfredini, Sergio Di Nuzzo, Shaniko Kaleci, Giovanni Pellacani and Francesca Farnetani

Diagnostics 2024, 14(5), 494; https://doi.org/10.3390/diagnostics14050494 - 25 Feb 2024

Viewed by 817

Abstract

Background: Reflectance confocal microscopy (RCM) has a defined in vivo morphology of vitiligo and re-pigmentation. Combination therapies seem more effective than monotherapies. Objective: We aim to describe the clinical and RCM features of re-pigmentation with combined narrowband ultraviolet B (NB-UVB) and piperine-based topical treatment in localized vitiligo. Methods: Eight patients enrolled at a single center received combined treatment: topical treatment was applied twice daily + NB-UVB twice weekly for 2 × 2-month periods. Clinical changes were analyzed by the Vitiligo Noticeability Scale (VNS) and percentage of re-pigmentation. The evaluator agreement was assessed. Predefined RCM features had the presence/absence of (i) blood vessels, (ii) dendritic cells, and the quantity of (i) an irregular honeycombed pattern and (ii) non-pigmented papillae. Clinical and RCM monitoring was performed at the baseline, 2, 3, 5, and 7 months. Results: Macules were “slightly less noticeable” with 25–50% re-pigmentation. Irregular honeycomb patterns and non-pigmented papillae were significantly less frequently observed, and in less extended areas (T1 vs. T2, p = 0.039; T0 vs. T1, p = 0.005 and T2 vs. T4, p = 0.033). Dendritic cells and blood vessels improved, with significant changes in blood vessels (T1 vs. T2, p = 0.005 and T3 vs. T4, p = 0.008). Conclusions: RCM confirms the morphological changes induced by combined treatment for localized vitiligo. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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10 pages, 2039 KiB

Open AccessArticle

Neuroinflammatory Findings of Corneal Confocal Microscopy in Long COVID-19 Patients, 2 Years after Acute SARS-CoV-2 Infection

by Pilar Cañadas, Leonela Gonzalez-Vides, Marta Alberquilla García-Velasco, Pedro Arriola, Noemí Guemes-Villahoz and Jose Luis Hernández-Verdejo

Diagnostics 2023, 13(20), 3188; https://doi.org/10.3390/diagnostics13203188 - 12 Oct 2023

Cited by 1 | Viewed by 3961

Abstract

Objective: To describe corneal confocal microscopy findings in patients with long COVID-19 with persistent symptoms over 20 months after SARS-CoV-2 infection. Design: A descriptive cross-sectional study that included a total of 88 patients; 60 patients with Long COVID-19 and 28 controls. Long COVID-19 diagnosis was established according to the World Health Organization criteria. Corneal confocal microscopy using a Heidelberg Retina Tomograph II (Heidelberg Engineering, Heidelberg, Germany) was performed to evaluate sub-basal nerve plexus morphology (corneal nerve fiber density, nerve fiber length, nerve branch density, nerve fiber total branch density, nerve fiber area, and nerve fiber width). Dendritic cell density and area, along with microneuromas and other morphological changes of the nerve fibers were recorded. Results: Long COVID-19 patients presented with reduced corneal nerve density and branch density as well as shorter corneal nerves compared to the control group. Additionally, Long COVID-19 patients showed an increased density of dendritic cells also with a greater area than that found in the control group of patients without systemic diseases. Microneuromas were detected in 15% of Long COVID-19 patients. Conclusions: Long COVID-19 patients exhibited altered corneal nerve parameters and increased DC density over 20 months after acute SARS-CoV-2 infection. These findings are consistent with a neuroinflammatory condition hypothesized to be present in patients with Long COVID-19, highlighting the potential role of corneal confocal microscopy as a promising noninvasive technique for the study of patients with Long COVID-19. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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13 pages, 2618 KiB

Open AccessArticle

The Multifaceted Role of Confocal Laser Endomicroscopy in Head and Neck Surgery: Oncologic and Functional Insights

by Nina Wenda, Kai Fruth, Annette Fisseler-Eckhoff and Jan Gosepath

Diagnostics 2023, 13(19), 3081; https://doi.org/10.3390/diagnostics13193081 - 28 Sep 2023

Cited by 1 | Viewed by 890

Abstract

(1) Background: Confocal laser endomicroscopy (CLE) has emerged as a transformative tool in head and neck surgery, with applications spanning oncologic insights and functional evaluations. This study delves into CLE’s potential in these domains. (2) Methods: We performed CLE in head and neck oncologic surgery, focusing on tumor margin identification and precise resection. We also employed CLE for functional assessment in allergic rhinitis, observing real-time mucosal changes during nasal provocation testing. (3) Results: In oncologic surgery, CLE enabled real-time visualization of tumor margins and cellular patterns, aiding resection decisions. In allergic rhinitis assessment, CLE captured dynamic morphological alterations upon allergen exposure, enhancing understanding of mucosal reactions. (4) Conclusions: The integration of CLE with evolving technologies such as deep learning and AI holds promise for enhanced diagnostic accuracy. This study underscores CLE’s expansive potential, highlighting its role in guiding surgical choices and illuminating inflammatory processes in the head and neck. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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10 pages, 967 KiB

Open AccessArticle

Socio-Organizational Impact of Confocal Laser Endomicroscopy in Neurosurgery and Neuropathology: Results from a Process Analysis and Expert Survey

by Marina L. Fotteler, Friederike Liesche-Starnecker, Maria C. Brielmaier, Johannes Schobel, Jens Gempt, Jürgen Schlegel and Walter Swoboda

Diagnostics 2021, 11(11), 2128; https://doi.org/10.3390/diagnostics11112128 - 16 Nov 2021

Viewed by 3436

Abstract

During brain tumor resection surgery, it is essential to determine the tumor borders as the extent of resection is important for post-operative patient survival. The current process of removing a tissue sample for frozen section analysis has several shortcomings that might be overcome by confocal laser endomicroscopy (CLE). CLE is a promising new technology enabling the digital in vivo visualization of tissue structures in near real-time. Research on the socio-organizational impact of introducing this new methodology to routine care in neurosurgery and neuropathology is scarce. We analyzed a potential clinical workflow employing CLE by comparing it to the current process. Additionally, a small expert survey was conducted to collect data on the opinion of clinical staff working with CLE. While CLE can contribute to a workload reduction for neuropathologists and enable a shorter process and a more efficient use of resources, the effort for neurosurgeons and surgery assistants might increase. Experts agree that CLE offers huge potential for better diagnosis and therapy but also see challenges, especially due to the current state of experimental use, including a risk for misinterpretations and the need for special training. Future studies will show whether CLE can become part of routine care. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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Review

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18 pages, 1358 KiB

Open AccessReview

Confocal Microscopy for Diagnosis and Management of Cutaneous Malignancies: Clinical Impacts and Innovation

by Mehmet Fatih Atak, Banu Farabi, Cristian Navarrete-Dechent, Gennady Rubinstein, Milind Rajadhyaksha and Manu Jain

Diagnostics 2023, 13(5), 854; https://doi.org/10.3390/diagnostics13050854 - 23 Feb 2023

Cited by 5 | Viewed by 2221

Abstract

Cutaneous malignancies are common malignancies worldwide, with rising incidence. Most skin cancers, including melanoma, can be cured if diagnosed correctly at an early stage. Thus, millions of biopsies are performed annually, posing a major economic burden. Non-invasive skin imaging techniques can aid in early diagnosis and save unnecessary benign biopsies. In this review article, we will discuss in vivo and ex vivo confocal microscopy (CM) techniques that are currently being utilized in dermatology clinics for skin cancer diagnosis. We will discuss their current applications and clinical impact. Additionally, we will provide a comprehensive review of the advances in the field of CM, including multi-modal approaches, the integration of fluorescent targeted dyes, and the role of artificial intelligence for improved diagnosis and management. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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19 pages, 3296 KiB

Open AccessReview

Development, Implementation and Application of Confocal Laser Endomicroscopy in Brain, Head and Neck Surgery—A Review

by Irini Kakaletri, Maximilian Linxweiler, Serine Ajlouni and Patra Charalampaki

Diagnostics 2022, 12(11), 2697; https://doi.org/10.3390/diagnostics12112697 - 04 Nov 2022

Cited by 6 | Viewed by 2298

Abstract

When we talk about visualization methods in surgery, it is important to mention that the diagnosis of tumors and how we define tumor borders intraoperatively in a correct way are two main things that would not be possible to achieve without this grand variety of visualization methods we have at our disposal nowadays. In addition, histopathology also plays a very important role, and its importance cannot be neglected either. Some biopsy specimens, e.g., frozen sections, are examined by a histopathologist and lead to tumor diagnosis and the definition of its borders. Furthermore, surgical resection is a very important point when it comes to prognosis and life survival. Confocal laser endomicroscopy (CLE) is an imaging technique that provides microscopic information on the tissue in real time. CLE of disorders, such as head, neck and brain tumors, has only recently been suggested to contribute to both immediate tumor characterization and detection. It can be used as an additional tool for surgical biopsies during biopsy or surgical procedures and for inspection of resection margins during surgery. In this review, we analyze the development, implementation, advantages and disadvantages as well as the future directions of this technique in neurosurgical and otorhinolaryngological disciplines. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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12 pages, 3804 KiB

Open AccessReview

The Use of Confocal Laser Endomicroscopy in Diagnosing Barrett’s Esophagus and Esophageal Adenocarcinoma

by Jitka Vaculová, Radek Kroupa, Zdeněk Kala, Jiří Dolina, Tomáš Grolich, Jakub Vlažný, David Said, Lydie Izakovičová Hollá, Petra Bořilová Linhartová, Vladimír Procházka, Marek Joukal, Petr Jabandžiev, Ondřej Slabý and Lumír Kunovský

Diagnostics 2022, 12(7), 1616; https://doi.org/10.3390/diagnostics12071616 - 02 Jul 2022

Cited by 1 | Viewed by 1935

Abstract

Confocal laser endomicroscopy (CLE) is a diagnostic technique that enables real-time microscopic imaging during microscopic examination and evaluation of epithelial structures with 1000-fold magnification. CLE can be used in the diagnosis of various pathologies, in pneumology, and in urology, and it is very widely utilized in gastroenterology, most importantly in the diagnosis of Barrett’s esophagus (BE), esophageal adenocarcinoma (EAC), biliary strictures, and cystic pancreatic lesions. A literature search was made in MEDLINE/PubMed and Google Scholar databases while focusing on diagnostics using CLE of BE and EAC. We then examined randomized and observational studies, systematic reviews, and meta-analyses relating to the utilization of CLE in BE and EAC diagnostics. Here, we discuss whether CLE can be a suitable diagnostic method for surveillance of BE. Even though many studies have proven that CLE increases diagnostic accuracy in detecting neoplastic transformation of BE, CLE is still not used as a standard diagnostic tool in BE surveillance due to a deficiency of scientific evidence. More studies and data are needed if CLE is to find a place as a new technique in BE surveillance. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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Other

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2 pages, 745 KiB

Open AccessInteresting Images

In Vivo View of a Reclining Demodex Mite in a Milia Cyst

by Katharine Hanlon, Meredith Thomley and Lilia Correa-Selm

Diagnostics 2023, 13(10), 1718; https://doi.org/10.3390/diagnostics13101718 - 12 May 2023

Viewed by 1125

Abstract

Demodex folliculorum and Demodex brevis are commonly present on facial skin and frequently noted via Reflectance Confocal Microscopy (RCM) examination. These mites inhabit follicles and are often seen in groups of two or more, although D. brevis is usually found as a solitary mite. When observed through RCM, they are typically present as refractile, round groupings seen on a transverse image plane inside the sebaceous opening, as they are vertically oriented, and their exoskeletons refract under near-infrared light. Inflammation may occur, leading to a variety of skin disorders; nonetheless, these mites are considered to be part of normal skin flora. a 59-year-old woman presented to our dermatology clinic for confocal imaging (Vivascope 3000, Caliber ID, Rochester, NY, USA) of a previously excised skin cancer for margin evaluation. She did not exhibit symptoms of rosacea or active inflammation of the skin. Incidentally, a solitary demodex mite was noted in a milia cyst nearby the scar. The mite appeared to be trapped in the keratin-filled cyst and was positioned horizontally to the image plane such that its entire body was captured in a coronal orientation as a stack. Demodex identification using RCM can provide clinical diagnostic value in the context of rosacea or inflammation; in our case, this solitary mite was thought to be part of the patient’s normal skin flora. Demodex are practically ubiquitous on the facial skin of older patients and are frequently noted during RCM examination; however, the orientation of the mite referenced herein is uncommon, allowing for a unique view of its anatomy. The use of RCM to identify demodex may become more routine as access to technology grows. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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8 pages, 2318 KiB

Open AccessCase Report

Corneal Confocal Microscopy Findings in Neuro Lyme Disease: A Case Report

by Pilar Cañadas, Montserrat García-Gonzalez, Rafael Cañones-Zafra and Miguel A. Teus

Diagnostics 2022, 12(2), 343; https://doi.org/10.3390/diagnostics12020343 - 29 Jan 2022

Viewed by 4804

Abstract

Neuro Lyme disease is caused by several bacteriae of the Borreliaceae family, such as Borrelia Miyamotoi. In late stages of illness, patients with Lyme disease may develop chronic neurologic symptoms such as cognitive disturbances or small fiber peripheral neuropathy. Confocal microscopy is a non-invasive method designed to evaluate the human cornea in vivo. Thus, all the corneal layers, including the cells and the sub-basal nerve plexus, can be easily visualized and analyzed. This is the first report of the morphology of small-fiber peripheral neuropathy analyzed by confocal microscopy in a patient diagnosed of neuro Lyme disease. The decrease in the number of unmyelinated sub-basal nerve fibers with abundant presence of dendritic cells (DC) in comparison with healthy corneas strongly supports the diagnosis of small fiber peripheral neuropathy in a case of neuroborreliosis disease. Full article

(This article belongs to the Special Issue Confocal Microscopy: Clinical Impacts and Innovation)

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