In Vivo Multiphoton Microscopy of Basal Cell Carcinoma

Balu, Mihaela; Zachary, Christopher B.; Harris, Ronald M.; Krasieva, Tatiana B.; König, Karsten; Tromberg, Bruce J.; Kelly, Kristen M.

doi:10.1001/jamadermatol.2015.0453

Cited by 107 publications

(98 citation statements)

References 23 publications

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“…Increased NADH fluorescence intensity, as compared to normal epithelium, has also been revealed in human head and neck tumors in vivo [85]. Similar results have also been obtained when studying human basal cell carcinoma in vivo [3]. All authors attribute high NADH fluorescence intensity to active metabolic processes in tumor cells.…”

Section: Redox Ratio As Metabolic Status Indicatorsupporting

confidence: 68%

“…Cofactors NADH and FAD which are present in the cell in an oxidized (NAD + , FAD) or reduced form (NADN, FADN 2 ) play the role of electron donors and acceptors in reactions of ATP formation [1][2][3][4][5][6][7].…”

Section: Cofactors Nadn and Fad And Their Role In Cellular Energy Metmentioning

confidence: 99%

“…There is equipment for two-photon microscopy and FlIM, approved for application in clinic, such as MPTflex and DermaInspect systems produced by Jenlab (Germany) [3,96,97].…”

Section: Nadh and Fad Fluorescence Lifetime In Energy Metabolism Evalmentioning

confidence: 99%

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Metabolic Imaging in the Study of Oncological Processes (Review)

Lukina¹,

Shirmanova²,

Sergeeva³

et al. 2016

Sovrem Tehnol Med

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There have been reviewed the main approaches to the study of energy metabolism in cancer cells, based on fluorescent imaging of NADH and FAD cofactors. The term "metabolic imaging" covers a range of modern fluorescent techniques for detecting NADH and FAD according to fluorescence intensity and/or lifetime. These cofactors play an important role in energy metabolism reactions acting as electron carriers and, being fluorescent, serve as a basis for metabolic process analysis in living cells and tissues without the use of additional coloring agents. Particular attention is paid to metabolic changes associated with carcinogenesis. Numerous examples of metabolic imaging application in cell cultures in vitro, animal and human tumors in vivo, as well as in patients' tumor biopsy samples have demonstrated its being highly demanded for biomedical research in the area of oncology.Key words: metabolic imaging; energy metabolism; fluorescence lifetime; redox ratio; NADH; FAD; oxidative phosphorylation; glycolysis; tumor cells.For contacts: Mariya M. lukina, e-mail: kuznetsova.m.m@yandex.ru Cells need energy to maintain homeostasis. All processes of cell activity, such as generating concentration gradients, cytoskeleton movement, DNA repair, transcription, translation and vesicular transport, are energy-dependent. Energy metabolism of normal cells is known to differ significantly from cell metabolism in case of a disease. Therefore, the metabolic status can serve as an indicator for diagnosis and visualization of response to pathological process treatment. Taking into account high incidence of oncological diseases, assessment of metabolic phenotype of tumor cells is particularly urgent.Development of optical visualization technologies has provided the possibility of noninvasive analysis of metabolic cofactors NADH and FAD in living tumor cells at high spatial resolution (up to several hundred Metabolic Imaging in the Study of Oncological Process nanometers) without using additional coloring agents and with no significant effect on biochemical and physiological condition of cells. The term "metabolic imaging" covers a range of modern fluorescent techniques which allow visualizing NADH and FAD according to their fluorescence intensity and/or lifetime.The present review characterizes the properties of energy metabolism in cancer cells, describes the two key approaches to the assessment of metabolic status: analysis of cofactor fluorescence intensity ratio (redox ratio) and their fluorescence lifetime. There have been presented numerous examples of metabolic imaging application in cell cultures in vitro, animal and human tumors in vivo, as well as in patients' tumor biopsy samples.

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Section: Redox Ratio As Metabolic Status Indicatorsupporting

confidence: 68%

Section: Cofactors Nadn and Fad And Their Role In Cellular Energy Metmentioning

confidence: 99%

See 1 more Smart Citation

Metabolic Imaging in the Study of Oncological Processes (Review)

Lukina¹,

Shirmanova²,

Sergeeva³

et al. 2016

Sovrem Tehnol Med

View full text Add to dashboard Cite

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“…In BCC, cellular nests resided in the dermis surrounded by excessive collagen/elastin stromal deposition (Fig. 4A, green arrows) (24), findings representing typical histological BCC presentation. The cellular morphological features were more highly variable among the melanomas than among the BCCs, with dendritic cells (melanocytes) appearing within the upper epidermal layers, haphazard cellular morphology, or cells invading the dermis (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In the case of melanoma detection, these criteria rely on quantitative manual tracing analysis of lesion-specific features of interest, such as dendrite density, and assessment of the endogenous TPEF NADH intensity variations (23). In the case of BCC lesions, detection is based on the qualitative demonstration of important diagnostic features, such as the presence of nests of basaloid cells in the papillary and upper reticular dermis (24). Other optical techniques, such as optical coherence tomography and confocal reflectance microscopy, also rely on specialized morphological criteria for skin lesion evaluation (25, 26).…”

Section: Introductionmentioning

confidence: 99%

Imaging mitochondrial dynamics in human skin reveals depth-dependent hypoxia and malignant potential for diagnosis

et al. 2016

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Active changes in mitochondrial structure and organization facilitate cellular homeostasis. Because aberrant mitochondrial dynamics are implicated in a variety of human diseases, their assessment is potentially useful for diagnosis, therapy, and disease monitoring. Because current techniques for evaluating mitochondrial morphology are invasive or necessitate mitochondria-specific dyes, their clinical translation is limited. We report that mitochondrial dynamics can be monitored in vivo, within intact human skin by relying entirely on endogenous two-photon–excited fluorescence from the reduced metabolic coenzyme nicotinamide adenine dinucleotide (NADH). We established the sensitivity of this approach with in vivo, fast temporal studies of arterial occlusion-reperfusion, which revealed acute changes in the mitochondrial metabolism and dynamics of the lower human epidermal layers. In vitro hypoxic-reperfusion studies validated that the in vivo outcomes were a result of NADH fluorescence changes. To demonstrate the diagnostic potential of this approach, we evaluated healthy and cancerous human skin epithelia. Healthy tissues displayed consistent, depth-dependent morphological and mitochondrial organization patterns that varied with histological stratification and intraepithelial mitochondrial protein expression. In contrast, these consistent patterns were absent in cancerous skin lesions. We exploited these differences to successfully differentiate healthy from cancerous tissues using a predictive classification approach. Collectively, these results demonstrate that our label-free, automated, near real-time assessments of mitochondrial organization—relying solely on endogenous contrast—could be useful for accurate, noninvasive in vivo diagnosis.

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Real-time Analysis of Skin Biopsy Specimens With 2-Photon Fluorescence Microscopy

et al. 2022

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IMPORTANCE Nonmelanoma skin cancers (NMSCs) are primarily diagnosed through paraffin section histologic analysis of skin biopsy specimens that requires days to weeks before a formal diagnosis is reported. Two-photon fluorescence microscopy (TPFM) has the potential for point-of-care diagnosis of NMSC and other dermatologic conditions, which could enable same-visit diagnosis and treatment.OBJECTIVE To demonstrate that TPFM imaging of NMSC can occur within minutes of obtaining biopsies and provide similar histological features to those of conventional histology and evaluate TPFM diagnostic performance with respect to conventional histology. DESIGN, SETTING, AND PARTICIPANTSThis comparative effectiveness pilot study examined 29 freshly excised biopsies from confirmed NMSC lesions in patients presenting for treatment. Biopsies underwent imaging immediately with TPFM on site at Rochester Dermatologic Surgery (Victor, New York) between October 2019 and August 2021. The imaged biopsies were subsequently submitted for paraffin histology to produce coregistered images. Twelve of these coregistered image pairs (41.4%) were used as a training set. Fifteen (51.7%) were used in a masked evaluation by a board-certified dermatopathologist. Two (6.9%) were excluded from the study before evaluation because they could not be coregistered.MAIN OUTCOMES AND MEASURES Sensitivity, specificity, and accuracy of TPFM for NMSC biopsies were evaluated compared with conventional histology.RESULTS Fourteen of the 15 biopsy specimens (93.3%) in the evaluation set were identically diagnosed with TPFM and paraffin histology. The TPFM had 100% sensitivity (95% CI, 48%-100%), 100% specificity (95% CI, 69%-100%), and 100% accuracy (95% CI, 78%-100%) for basal cell carcinoma diagnosis. For squamous cell carcinoma diagnosis, TPFM had 89% sensitivity (95% CI, 52%-100), 100% specificity (95% CI, 54%-100%), and 93% accuracy (95% CI, 68%-100%). For overall NMSC diagnosis, TPFM had a 93% sensitivity (95% CI, 66%-100%), 100% specificity (95% CI, 3%-100%), and 93% accuracy (95% CI, 68%-100%). Examination of the 1 discordant pair revealed mismatched imaging planes as the source of error. CONCLUSIONS AND RELEVANCEThe results of this comparative effectiveness pilot study suggest that TPFM captures histological characteristics of NMSC that are present in conventional histology, which reveals its potential as a rapid, point-of-care diagnostic alternative that does not need extensive sample preparation or retraining for image evaluation. Further validation of TPFM imaging performed for a larger cohort is needed to fully evaluate its diagnostic accuracy and potential effect within the field.

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In Vivo Multiphoton Microscopy of Basal Cell Carcinoma

Cited by 107 publications

References 23 publications

Metabolic Imaging in the Study of Oncological Processes (Review)

Metabolic Imaging in the Study of Oncological Processes (Review)

Imaging mitochondrial dynamics in human skin reveals depth-dependent hypoxia and malignant potential for diagnosis

Real-time Analysis of Skin Biopsy Specimens With 2-Photon Fluorescence Microscopy

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