ABSTRACT
BACKGROUND: Mucous membranes may be affected by various diseases and the diagnostic accessibility is limited. Multiphoton laser tomography (MPT) is a useful tool for in vivo evaluation of superficial skin structures and might also be useful for the imaging of mucosa. OBJECTIVES: In order to investigate the suitability of MPT for the evaluation of mucous membranes, tissue samples of different donors and anatomical localizations have been imaged. METHODS: Human mucosa samples from the urinary bladder, palatine tonsil and ocular conjunctiva were investigated by MPT and subsequently compared with conventional histology. RESULTS: Horizontal images of the epithelium and the underlying connective tissue were obtained using the autofluorescence and second harmonics generation signals by MPT. The stratification of multilayered epithelium was consistently reproduced. Also inflammatory changes like lymphocytic infiltrates and widened intercellular spaces were imaged in some cases. CONCLUSION: Mucous tissue samples were obtained and MPT imaging was successfully performed. In the direct comparison with histology of the same tissue samples, the characteristic morphology of the epithelia was found. Further studies are necessary in order to compare images of healthy and diseased mucosal epithelium and the use of MPT for its evaluation in vivo.
Subject(s)
Conjunctiva/anatomy & histology , Mucous Membrane/anatomy & histology , Palatine Tonsil/anatomy & histology , Urinary Bladder/anatomy & histology , Humans , Lasers , Mucous Membrane/pathology , Palatine Tonsil/pathology , TomographyABSTRACT
BACKGROUND: Multiphoton laser tomography (MPT) is a non-invasive technique that allows imaging of skin in vivo with very high spatial resolution and contrast. Previous work of our group has demonstrated that known morphological changes due to erythematogenic ultraviolet B (UVB) irradiation may be imaged in vivo by MPT. The present work investigated if morphological skin changes known from experimental erythematogenic UVB irradiation are also demonstrable in the course of a standard phototherapy regime that implies suberythematogenic doses of narrow band UVB. METHODS: Sixteen patients with psoriasis vulgaris receiving a narrow band phototherapy were included. A test field and a light-protected control field were measured with the multiphoton tomograph DermaInspect® at four time points: at baseline, the next day, after 3 days and at the day of the last exposure. RESULTS: In the course of the UVB phototherapy, spongiosis and pleomorphy as parameters of inflammation and cellular damage did not show significant changes. By contrast, an adaptive skin reaction with significant changes of keratosis and pigmentation was observed. CONCLUSION: MPT is a suitable technique for the investigation of qualitative and quantitative skin changes after UVB irradiation. After suberythematogenic UVB irradiation, photoadaptive skin changes, but no cellular damage can be observed with MPT.
Subject(s)
Epidermis/diagnostic imaging , Epidermis/radiation effects , Microscopy, Fluorescence, Multiphoton/methods , Psoriasis/diagnostic imaging , Psoriasis/radiotherapy , Ultraviolet Therapy/methods , Adult , Aged , Epidermis/pathology , Female , Humans , Lasers , Male , Middle Aged , Psoriasis/pathology , Tomography, Optical/methods , Treatment Outcome , Young AdultABSTRACT
In recent years, multiphoton tomography (MPT) and multiphoton microscopy have gained increasing importance as noninvasive examination techniques in dermatology. MPT imaging is based on the specific stimulation of biogenic fluorophores. The induction of second harmonic generation is also used for imaging of particular molecules. Additional fluorescence staining or fluorescence markers are not necessary-an important advantage for the in vivo examination of human skin. Multiphoton techniques are not only appropriate for clinical diagnostics but also for biomedical research. MPT provides an optical biopsy depth up to 200 µm with subcellular resolution depicting cellular and extracellular structures. In combination with fluorescence lifetime imaging, additional information about the microenvironment, the energetic state and the cellular metabolism can be obtained. This review presents recent developments of MPT for the in vivo evaluation of physiological and pathological changes of skin and diagnostics of dermal diseases.
Subject(s)
Microscopy, Fluorescence, Multiphoton/methods , Molecular Imaging/methods , Skin Diseases/metabolism , Skin Diseases/pathology , Skin/metabolism , Skin/pathology , Biomarkers/metabolism , Dermoscopy/methods , Humans , Image Enhancement/methods , Image Interpretation, Computer-Assisted/methodsABSTRACT
BACKGROUND: In vivo multiphoton tomography (MPT) of human skin has become a valuable tool for non-invasive examination of morphological and biophysical skin properties and their alterations. So far, skin changes after UVB irradiation were mainly evaluated clinically and histologically. The present study aimed at non-invasive imaging of histological changes during acute UVB irradiation by multiphoton laser tomography. METHODS: In 10 volunteers, five areas were irradiated once with an erythematous UVB dose. Multiphoton measurements were performed four times, i.e. before irradiation (baseline), and 24, 48 and 72 h after irradiation, respectively. The data were evaluated for changes of epidermal pleomorphy, spongiosis, pigmentation and thickness. RESULTS: The four parameters were altered significantly by acute UVB irradiation, i.e. epidermal pleomorphy, spongiosis, pigmentation and thickness increased within 72 h after irradiation. CONCLUSION: Thus, the study has shown that typical epidermal changes induced by acute UVB irradiation can be evaluated by MPT.
Subject(s)
Epidermis/injuries , Epidermis/pathology , Erythema/pathology , Microscopy, Fluorescence, Multiphoton/methods , Radiation Injuries/pathology , Ultraviolet Rays/adverse effects , Adult , Aged , Dose-Response Relationship, Drug , Epidermis/radiation effects , Erythema/etiology , Female , Humans , Lasers , Male , Middle Aged , Radiation Dosage , Radiation Injuries/etiology , Tomography, Optical/methods , Young AdultABSTRACT
BACKGROUND: Multiphoton tomography (MPT) is a novel non-invasive imaging method in dermatology allowing the depiction of the epidermis with sub-cellular resolution. Here, we present a descriptive characterization of unaffected human epidermis, morphometric data on human keratinocytes and some epidermal parameters in vivo and a morphological characterization of keratinocyte changes in actinic keratoses. METHODS: In a clinical setting, 57 volunteers of different age groups were examined using MPT. RESULTS: The morphological appearance of keratinocytes showed polygonal cells in the horny layer, a granular cytoplasm in the stratum granulosum, smaller prickle cells in the stratum spinosum and hyperpigmented small round basal cells. Actinic keratoses presented remarkable differences including widened inter-cellular spaces, heterogeneity in cellular fluorescence and shape as well as an increased ratio of nuclear to cellular size. Finally, the thickness of the epidermis was significantly increased in actinic keratoses compared with the control. CONCLUSION: In vivo MPT provides high-resolution images allowing the identification and quantification of cellular morphometric parameters. First observations of morphology and morphometry of actinic keratoses are reported.
Subject(s)
Dermoscopy/methods , Keratinocytes/cytology , Keratosis, Actinic/pathology , Microscopy, Fluorescence, Multiphoton/methods , Tomography/methods , Adult , Aged , Aged, 80 and over , Dermis/pathology , Epidermis/pathology , Female , Humans , Male , Microscopy, Confocal/methods , Middle Aged , Skin Aging/pathology , Young AdultABSTRACT
Over the last decades, there has been a dramatic increase of skin tumour incidence that results from actinic damage. This increase is mainly due to the current demographic development, but also due to a change in spare time habits. Thus, the treatment and prevention of light induced skin tumours is gaining interest, but also the treatment of photoaged skin. This review will focus on 1) the basic principles of actinic damage including carcinogenesis; 2) treatment options for photoaged skin and 3) an overview of current therapy concepts of light induced skin tumours.
Subject(s)
Skin Diseases/etiology , Skin Diseases/therapy , Skin Neoplasms/therapy , Sunlight/adverse effects , Humans , Keratosis, Actinic/etiology , Keratosis, Actinic/therapy , Skin Aging , Skin Neoplasms/etiologyABSTRACT
RegRand (Version 1.0) is Macintosh-based software that enables a researcher to conduct a non-parametric statistical analysis of the data from Koehler and Levin's (1998) recently described regulated randomization single-case multiple-baseline design. Regulated randomization design and analysis principles are reviewed in relation to an educational research application and a step-by-step illustration of them in relation to the RegRand program is presented.
