Fibre optic confocal imaging (FOCI) enabled subsurface fluorescence microscopy of the skin of hairless mice in vivo. Application of acridine orange enabled imaging of the layers of the epidermis. The corneocytes of the stratum corneum, the keratinocytes in the basal layers and redundant hair follicles were visualised at depths greater than 100 µm. Cellular and nuclear membranes of keratinocytes of the skin were visualised by the use of acridine orange and DIOC5(3). Imaging of the skin after injection of FITC-dextran revealed an extensive network of blood vessels with a size range up to 20 µm. Blood cells could be seen moving through dermal vessels and the blood circulation through the dermal vascular bed was video-taped. The fluorescent dye 4-di-2-ASP showed the presence of nerves fibres around the hair follicles and subsurface blood vessels. Comparison was made between images obtained in vivo using FOCI and in vitro scanning electron microscopy and conventional histology. FOCI offers the potential to study dynamic events in vivo, such as blood flow, skin growth, nerve regeneration and many pathological processes, in ways which have not previously been possible.

A fluorescence confocal microscopy technique was employed to obtain subsurface images of nerve and microvascular structure in the vas deferens and colon of the living rat. The use of dual labelling with vital dyes and 2-channel confocal acquisition allowed differentiation of microscopic structure at both low and higher magnification. Characteristic staining patterns of nerves and blood vessels were repeatedly obtained in each tissue, suggesting the potential of this technique for studying morphological changes associated with surgical procedures and\or models of neuronal or vascular pathology.

Purpose Certain types of repetitive sub-maximal knee loading cause microfatigue damage in the human anterior cruciate ligament (ACL) that can accumulate to produce macroscopic tissue failure. However, monitoring the progression of that ACL microfatigue damage as a function of loading cycles has not been reported. To explore the fatigue process, a confocal laser endomicroscope (CLEM) was employed to capture sub-micron resolution fluorescence images of the tissue in situ. The goal of this study was to quantify the in situ changes in ACL autofluorescence (AF) signal intensity and collagen microstructure as a function of the number of loading cycles. Methods Three paired and four single cadaveric knees were subjected to a repeated 4 times bodyweight landing maneuver known to strain the ACL. The paired knees were used to compare the development of ACL microfatigue damage on the loaded knee after 100 consecutive loading cycles, relative to the contralateral unloaded control knee, through second harmonic generation (SHG) and AF imaging using confocal microscopy (CM). The four single knees were used for monitoring progressive ACL microfatigue damage development by AF imaging using CLEM. Results The loaded knees from each pair exhibited a statistically significant increase in AF signal intensity and decrease in SHG signal intensity as compared to the contralateral control knees. Additionally, the anisotropy of the collagen fibers in the loaded knees increased as indicated by the reduced coherency coefficient. Two out of the four single knee ACLs failed during fatigue loading, and they exhibited an order of magnitude higher increase in autofluorescence intensity per loading cycle as compared to the intact knees. Of the three regions of the ACL - proximal, midsubstance and distal - the proximal region of ACL fibers exhibited the highest AF intensity change and anisotropy of fibers. Conclusions CLEM can capture changes in ACL AF and collagen microstructures in situ during and after microfatigue damage development. Results suggest a large increase in AF may occur in the final few cycles immediately prior to or at failure, representing a greater plastic deformation of the tissue. This reinforces the argument that existing microfatigue damage can accumulate to induce bulk mechanical failure in ACL injuries. The variation in fiber organization changes in the ACL regions with application of load is consistent with the known differences in loading distribution at the ACL femoral enthesis.

ObjectiveTo systematically identify and summarise current research on the utility of confocal microscopy in oral squamous cell carcinoma and oral epithelial dysplasia in oral potentially malignant disorders.MethodsDatabases Medline, Embase, Evidence‐Based Medicine, and Web of Science were searched with articles screened and included if their primary objective was the use of a confocal microscope in diagnosis of oral cancer or epithelial dysplasia, in vivo or ex vivo.Results and DiscussionTwenty‐eight relevant studies were identified of which 21 studies included oral squamous cell carcinoma specimens. Fifteen studies included in vivo use. The studies included both qualitative and fluorescence confocal microscope and reflectance confocal microscope analysis along with quantitative analysis of carcinoma and dysplasia. Thirteen studies reported the predictive value of their confocal device in the diagnosis of dysplasia and carcinoma. The quantitative software‐based studies show promise in objectifying the diagnostic process for identifying abnormalities within the microstructure of the oral mucosa.ConclusionsThere was heterogeneity in the criteria for diagnosis of dysplasia and oral squamous cell carcinoma with experience levels of assessors impacting method efficacy. Both qualitative and quantitative confocal assessment methodologies have been explored, the latter highlighting the potential of future machine‐augmented diagnostic precision.

1. The tetrabrominated diphenyl ether 3,5-dibromo-2-(2,4-dibromophenoxy)phenol (BPE), a natural marine product isolated from a sponge, was tested for pharmacological activity in guinea-pig ileum. 2. BPE (2 mumol/L) decreased basal force and the frequency of spontaneous contractions of the ileum. It also significantly decreased contractions of the ileum induced by 5 mmol/L barium and to electrical stimulation at parameters which stimulated intrinsic nerves. 3. The slopes of concentration-response curves to acetylcholine (ACh), histamine and 5-hydroxytryptamine (5-HT) were significantly reduced by BPE at concentrations of 2 mumol/L or greater. 4. BPE (2 mumol/L) did not affect calcium-induced contractions of longitudinal muscle fibres from guinea-pig ileum which were stripped of their cellular membrane. It (6 mumol/L) also had no effect on ATP levels in longitudinal muscle fibres. 5. BPE (2 mumol/L) reduced both phasic and tonic components of contractions induced by raising the extracellular concentration of K+ to 15, 30, 45 or 60 mmol/L (in the presence of atropine, propranolol, phentolamine and desensitization to 5-HT to inhibit the effects of nerve transmitter release). 6. BPE (2 mumol/L) reduced carbachol-induced contractions of ileum pre-incubated in 1 mumol/L felodipine, a blocker of L-type voltage-operated calcium channels (VOCC). 7. BPE dose dependently (0.6-6 mumol/L) reduced contractions induced by Ca2+ in both K+ depolarized ileum and in tissue exposed to carbachol (10 mumol/L) in the presence of felodipine (0.1 mumol/L). 8. These results suggest that BPE affects intracellular messenger systems controlling cytosolic calcium and/or blocks entry of calcium into the cell through both VOCC and receptor-operated channels (ROC).