Precise removal of basal cell carcinomas (BCCs) with minimal damage to the surrounding normal skin is guided by the examination of frozen histology of each excision during Mohs surgery. The preparation of frozen histology is slow, requiring 20 to 45 min per excision. Confocal reflectance mosaicing may enable rapid detection of BCCs directly in surgical excisions, with minimal need for frozen histology. Soaking the excisions in acetic acid rapidly brightens nuclei and enhances BCC-to-dermis contrast. Clinically useful concentrations of acetic acid from 10 to 1% require 30 s to 5 min, respectively. A tissue fixture precisely controls the stability, flatness, tilt, and sag of the excisions, which enables mosaicing of 36x36 images to create a field of view of 12x12 mm. This simulates a 2x magnification view in light microscopes, which is routinely used by Mohs surgeons to examine frozen histology. Compared to brightfield, cross-polarization enhances contrast and detectability of BCCs in the papillary dermis but not in the reticular dermis. Comparison of mosaics to histology shows that nodular, micronodular, and superficial BCCs are easily detected. However, infiltrative and sclerosing BCCs tend to be obscured within the surrounding bright dermis. The mosaicing method currently requires 9 min, and thus may expedite Mohs surgery.
SummaryPrecise micro-surgical removal of tumour with minimal damage to the surrounding normal tissue requires a series of excisions, each guided by an examination of frozen histology of the previous. An example is Mohs surgery for the removal of basal cell carcinomas (BCCs) in skin. The preparation of frozen histology is labour-intensive and slow. Confocal microscopy may enable rapid detection of tumours directly in surgical excisions with minimal need for frozen histology. Mosaicing of images enables observation of nuclear and cellular morphology in large areas of surgically excised tissue. In skin, the use of 10-1% acetic acid as a reflectance contrast agent brightens nuclei in 0.5-5 min and enhances nuclear-to-dermis contrast and detectability of BCCs. A tissue fixture was engineered for precisely mounting surgical excisions to enable mosaicing of 36 × 36 images to create a field of view of 12 × 12 mm. This large field of view displays the excision at 2× magnification, similar to that routinely used by Mohs surgeons when examining frozen histology. Comparison of mosaics to histology demonstrates detectability of BCCs. Confocal mosaicing presently requires 9 min, instead of 20-45 min per excision for preparing frozen histology, and thus may provide a means for rapid pathology-at-the-bedside to expedite and guide surgery.
Aim:This study was conducted to evaluate the impact of summer and winter season on physiological, biochemical, hormonal, and antioxidant parameters in Indigenous sheep.Materials and Methods:The research was carried out during summer and winter season. 8 adult apparently healthy female sheep (aged 2-4 years) of similar physiological status were selected. Daily ambient temperature and relative humidity were recorded to calculate the temperature-humidity index (THI). The THI value of summer and winter season were 82.55 and 59.36, respectively, which indicate extreme hot condition during summer season and extreme cold condition during winter season. Physiological parameters were recorded daily during the experimental periods. Blood samples were collected at weekly interval and analyzed for biochemical, hormonal, and antioxidant parameters. The results were analyzed using completely randomized design.Results:From data obtained in this study, we found that higher THI during summer have significant effect over various physiological, biochemical, hormonal, and enzymatic indices of indigenous sheep. The physiological response such as rectal temperature, respiration rate (RR), pulse rate (PR), and skin temperature (ST) was increased significantly. We also found a significant increase in some biochemical parameters such as blood urea nitrogen (BUN), uric acid, creatinine (Cr), alanine transaminase (ALT), aspartate transaminase (AST), sodium (Na), and potassium (K). The level of cortisol hormone and superoxide dismutase (SOD), glutathione peroxidase (GPx), and lipid peroxidase (LPO) antioxidants increased significantly during summer. Whereas, some parameters such as glucose, cholesterol, calcium (Ca), inorganic phosphorus (IP), triiodothyronine (T3), and thyroxine (T4) were decreased significantly during summer season.Conclusion:It was concluded that the THI is a sensitive indicator of heat stress and is impacted by ambient temperature more than the relative humidity in Indigenous sheep. Higher THI is associated with significant increase in RT, RR, PR, ST, BUN, uric acid, Cr, ALT, AST, Na, K, cortisol, SOD, GPx, and LPO and with a significant decrease in glucose, cholesterol, Ca, IP, T3 and T4.
Both point-scanning and line-scanning confocal microscopes provide resolution and optical sectioning to observe nuclear and cellular detail in human tissues, and are being translated for clinical applications. While traditional point-scanning is truly confocal and offers the best possible optical sectioning and resolution, line-scanning is partially confocal but may offer a relatively simpler and lower-cost alternative for more widespread dissemination into clinical settings. The loss of sectioning and loss of contrast due to scattering in tissue is more rapid and more severe with a line-scan than with a point-scan. However, the sectioning and contrast may be recovered with the use of a divided-pupil. Thus, as part of our efforts to translate confocal microscopy for detection of skin cancer, and to determine the best possible approach for clinical applications, we are now developing a quantitative understanding of imaging performance for a set of scanning and pupil conditions. We report a Fourier-analysis-based computational model of confocal microscopy for six configurations. The six configurations are point-scanning and line-scanning, with full-pupil, half-pupil and divided-pupils. The performance, in terms of on-axis irradiance (signal), resolution and sectioning capabilities, is quantified and compared among these six configurations.
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