Texaphyrin-Based Calcium Sensor for Multimodal Imaging.

The ability to monitor intracellular calcium concentrations using fluorescent probes has led to important insights into biological signaling processes at the cellular level. An important challenge is to relate such measurements to broader patterns of signaling across fields of view that are inaccessible to optical techniques. To meet this need, we synthesized molecular probes that couple calcium-binding moieties to lanthanide texaphyrins, resulting in complexes endowed with a diverse complement of magnetic and photophysical properties. We show that the probes permit intracellular calcium levels to be assessed by fluorescence, photoacoustic, and magnetic resonance imaging modalities and that they are detectable by multimodal imaging in brain tissue. This work thus establishes a route for monitoring signaling processes over a range of spatial and temporal scales.

Mapping light distribution in tissue by using MRI-detectable photosensitive liposomes.

Characterizing sources and targets of illumination in living tissue is challenging. Here we show that spatial distributions of light in tissue can be mapped by using magnetic resonance imaging (MRI) in the presence of photosensitive nanoparticle probes. Each probe consists of a reservoir of paramagnetic molecules enclosed by a liposomal membrane incorporating photosensitive lipids. Incident light causes the photoisomerization of the lipids and alters hydrodynamic exchange across the membrane, thereby affecting longitudinal relaxation-weighted contrast in MRI. We injected the nanoparticles into the brains of live rats and used MRI to map responses to illumination profiles characteristic of widely used applications of photostimulation, photometry and phototherapy. The responses deviated from simple photon propagation models and revealed signatures of light scattering and nonlinear responsiveness. Paramagnetic liposomal nanoparticles may enable MRI to map a broad range of optical phenomena in deep tissue and other opaque environments.

Caries inhibition of simulated active caries lesions with CO2 laser irradiation and fluoride.

It has been well established that CO2 laser irradiation can be used to transform the mineral phase of dental enamel to make it more resistant to acid dissolution. The purpose of this study was to investigate if carbon dioxide laser irradiation and topical fluoride can be used to treat incipient caries lesions to inhibit further progression, i.e. treat active lesion surfaces as opposed to sound surfaces prior to subjecting them to an acid challenge. Simulated active caries lesions were produced on twenty eight bovine enamel samples using a pH cycling model and those surfaces were irradiated by a 9.4 µm CO2 laser and treated with topical fluoride. Changes in the surface morphology, acid resistance, and permeability were measured using digital microscopy, optical coherence tomography (OCT), and SWIR reflectance surface dehydration rate measurements at 1950 nm after exposure to a further acid challenge. There was a significant reduction (P < 0.05) of further lesion progression for lesion windows treated with CO2 laser irradiation followed by the application of an acidulated phosphate fluoride gel compared to the untreated lesion windows on each sample. Treatment by laser irradiation alone was not effective. The degree of lesion inhibition was not as high as has been previously observed for laser irradiated sound enamel surfaces exposed to an acid challenge.

Efficacy of Pre-Procedural Mouthwashes against SARS-CoV-2: A Systematic Review of Randomized Controlled Trials.

The oral mucosa is one of the first sites to be affected by the SARS-CoV-2. For this reason, healthcare providers performing aerosol-generating procedures (AGPs) in the oral cavity are at high risk of infection with COVID-19. The aim of this systematic review is to verify whether there is evidence in the literature describing a decrease in the salivary viral load of SARS-CoV-2 after using different mouthwashes. An electronic search of the MEDLINE database (via PubMed), Web of Science, SCOPUS, and the Cochrane library database was carried out. The criteria used were those described by the PRISMA® Statement. Randomized controlled trial studies that have used mouthwashes as a form of intervention to reduce the viral load in saliva were included. The risk of bias was analyzed using the Joanna Briggs Institute Critical Appraisal Tool. Ultimately, eight articles were included that met the established criteria. Based on the evidence currently available in the literature, PVP-I, CHX and CPC present significant virucidal activity against SARS-CoV-2 in saliva and could be used as pre-procedural mouthwashes to reduce the risk of cross-infection.

High contrast imaging of dental fluorosis in the short wavelength infrared.

Dental fluorosis is an increasing problem due to over exposure to fluoride from the environment. Fluorosis causes hypomineralization of the enamel during tooth development and mild fluorosis is visible as faint white lines on the tooth surface while the most severe fluorosis can result in pitted surfaces. It is difficult to quantify the severity of mild to moderate fluorosis and assessments are limited to subjective visual examinations. Dental fluorosis appears with very high contrast at short wavelength infrared (SWIR) wavelengths beyond 1400 nm and we hypothesize that these wavelengths may be better suited for detecting mild fluorosis and for estimating the severity on tooth surfaces. In this study, the contrast of fluorosis of varying severity on extracted human permanent teeth was measured at SWIR wavelengths ranging from 1300 to 2150 nm using an extended range of InGaAs camera and broadband light sources. The contrast was also measured in the visible range and with quantitative light-induced fluorescence (QLF) for comparison. The depth of hypomineralization and the integrated reflectivity were also measured with cross-polarization optical coherence tomography. The contrast of hypomineralization is significantly higher (P < 0.05) at 1460 and 1950 nm wavelengths than for the visible, fluorescence or other SWIR wavelengths from 1300 to 2150 nm. The highest correlation of the contrast with the depth of hypomineralization measured with cross-polarization-optical coherence tomography (CP-OCT) was at 1950 nm. This SWIR in vitro imaging study exploring wavelengths beyond 1400 nm has shown that hypomineralization on tooth surfaces can be viewed with extremely high contrast at SWIR wavelengths from 1460 to 2000 nm and that SWIR imaging has great potential for monitoring hypomineralization on tooth surfaces. New clinical methods are needed for the measurement of fluorosis that are valid, reliable, and feasible for surveillance at the community level. In addition, methods are needed for the quantitative assessment of fluorosis in vivo.

Computer-Controlled CO2 Laser Ablation System for Cone-beam Computed Tomography and Digital Image Guided Endodontic Access: A Pilot Study.

INTRODUCTION: Ideal endodontic access provides unobstructed entry to the pulp chamber and visualization of the canal orifices while preserving the maximum amount of tooth structure. The aim of this study was to implement the use of lasers to accurately and predictably access teeth to follow the principles of minimally invasive endodontics. METHODS: Traditional, conservative, ultraconservative, bridge, truss, and orifice-directed accesses were performed. A computer-controlled 9.3-µm CO2 laser ablation system was assembled and coupled with custom software capable of combining cone-beam computed tomographic (CBCT) volumetric data with spatially calibrated digital images of teeth to provide an augmented reality environment for designing and preparing endodontic accesses. Twenty (N = 20) sound posterior teeth with fully developed root canal systems were imaged with CBCT scans and accessed via laser ablation in vitro. RESULTS: All 20 (20/20) teeth were successfully accessed without iatrogenic errors. Volumetric renderings from post-access CBCT scans were used to verify the access and determine accuracy qualitatively. The volumetric measurements of hard tissue removed were as follows: traditional = 39.41 mm3, conservative = 9.76 mm3, ultraconservative = 7.1 mm3, bridge = 11.53 mm3, truss = 19.21 mm3, and orifice directed = 16.86 mm3. CONCLUSIONS: Digital image guidance based on feature recognition and registration with CBCT data is a viable method to address the challenge of dynamic navigation for accessing the pulp chamber. Modern lasers with high pulse repetition rates integrated with computer-controlled scanning systems are suitable for the efficient cutting of dental hard tissues.

Compact in vivo handheld dual SWIR transillumination/reflectance imaging system for the detection of proximal and occlusal lesions.

We have developed a clinical probe capable of acquiring simultaneous short wavelength infrared (SWIR) cross-polarized reflectance and occlusal transillumination images of lesions on tooth proximal and occlusal surfaces. We hypothesize that the dual SWIR reflectance and transillumination probe will improve the diagnostic accuracy of the device by reducing false positives since it is unlikely that confounding structural features or specular reflection are going to be present in both reflectance and transillumination images. In addition, the dual probe will provide complementary diagnostic information about lesion severity to help discriminate early superficial lesions on tooth surfaces from deeply penetrating lesions. The dual probe was 3D printed and equipped with a compact InGaAs camera and broadband superluminescent diode light sources that emit broadband light at 1300 nm for occlusal transillumination and 1600 nm light for cross-polarization reflectance measurements. The first clinical images acquired using this novel probe are presented.

Combined effects of a ketogenic diet and exercise training alter mitochondrial and peroxisomal substrate oxidative capacity in skeletal muscle.

Ketogenic diets (KDs) are reported to improve body weight, fat mass, and exercise performance in humans. Unfortunately, most rodent studies have used a low-protein KD, which does not recapitulate diets used by humans. Since skeletal muscle plays a critical role in responding to macronutrient perturbations induced by diet and exercise, the purpose of this study was to test if a normal-protein KD (NPKD) impacts shifts in skeletal muscle substrate oxidative capacity in response to exercise training (ExTr). A high fat, carbohydrate-deficient NPKD (16.1% protein, 83.9% fat, 0% carbohydrate) was given to C57BL/6J male mice for 6 wk, whereas controls (Con) received a low-fat diet with similar protein (15.9% protein, 11.9% fat, 72.2% carbohydrate). After 3 wk on the diet, mice began treadmill training 5 days/wk, 60 min/day for 3 wks. The NPKD increased body weight and fat mass, whereas ExTr negated a continued rise in adiposity. ExTr increased intramuscular glycogen, whereas the NPKD increased intramuscular triglycerides. Neither the NPKD nor ExTr alone altered mitochondrial content; however, in combination, the NPKD-ExTr group showed increases in PGC-1α and markers of mitochondrial fission/fusion. Pyruvate oxidative capacity was unchanged by either intervention, whereas ExTr increased leucine oxidation in NPKD-fed mice. Lipid metabolism pathways had the most notable changes as the NPKD and ExTr interventions both enhanced mitochondrial and peroxisomal lipid oxidation and many adaptations were additive or synergistic. Overall, these results suggest that a combination of a NPKD and ExTr induces additive and/or synergistic adaptations in skeletal muscle oxidative capacity.NEW & NOTEWORTHY A ketogenic diet with normal protein content (NPKD) increases body weight and fat mass, increases intramuscular triglyceride storage, and upregulates pathways related to protein metabolism. In combination with exercise training, a NPKD induces additive and/or synergistic activation of AMPK, PGC-1α, mitochondrial fission/fusion genes, mitochondrial fatty acid oxidation, and peroxisomal adaptations in skeletal muscle. Collectively, results from this study provide mechanistic insight into adaptations in skeletal muscle relevant to keto-adaptation.

Imaging dental fluorosis at SWIR wavelengths from 1300 to 2000-nm.

Dental fluorosis is an increasing problem in the U.S. due to excessive exposure to fluoride from the environment. Fluorosis causes hypomineralization of the enamel during tooth development and mild fluorosis is visible as faint white lines on the tooth surface while the most severe fluorosis can result in pitted surfaces. It is difficult to quantify the severity of fluorosis and assessments are limited to subjective visual assessments. Dental fluorosis appears with very high contrast at short wavelength infrared (SWIR) wavelengths beyond 1400-nm and we hypothesize that these wavelengths may be better suited for detecting mild fluorosis and for estimating the severity. In this study the contrast of fluorosis of varying severity on extracted human permanent teeth was measured at SWIR wavelengths ranging from 1300-2000-nm using an extended range InGaAs camera and broadband light sources. Cross polarization optical coherence tomography was used to measure the depth of hypomineralization.

Multispectral SWIR images of the pulp-chamber of posterior teeth in vitro.

Intraoral imaging of teeth with SWIR light provides increased contrast of dental caries and restorative materials compared to visible inspection and digital radiography. The objective of this study was to investigate the SWIR optical properties of the dental pulp-chamber floor, walls and canal orifices. We imaged in vitro extracted human posterior teeth at 1300-nm and 1500-1700-nm in reflectance and transillumination and compared the tissues properties with visible light images and quantitative light fluorescence. Transillumination of posterior teeth at both 1300-nm and 1500-1700-nm yielded significantly higher contrast between the pulp-chamber floor and walls than all other methods tested.

Dual short wavelength infrared transillumination/reflectance mode imaging for caries detection.

SIGNIFICANCE: We have developed a clinical probe capable of acquiring near-simultaneous short-wavelength infrared (SWIR) reflectance and occlusal transillumination images of lesions on tooth proximal and occlusal surfaces. We hypothesize that dual images will aid in differentiating between shallow and deep occlusal lesions and reduce the potential of false positives (FPs). AIM: The aim of this study was to test the performance of the dual reflectance and occlusal transillumination probe on extracted teeth prior to commencing clinical studies. APPROACH: The dual probe was 3D printed and the imaging system uses an InGaAs camera and broadband superluminescent diode light sources that emit broadband light at 1300 nm for occlusal transillumination and 1600-nm light for cross-polarization reflectance. The diagnostic performance of the dual probe was assessed using 120 extracted teeth with approximal and occlusal lesions. Reflectance and transillumination images were fused into single images to enhance the contrast between sound and lesion areas. The lesion contrast in both modes did not increase significantly with either the lesion depth or the distance from the occlusal surface for approximal lesions. In addition, the diagnostic performance of radiography, the individual reflectance and transillumination images, dual images, and fused images were compared using micro-computed tomography as the gold standard. RESULTS: Reflectance imaging at 1600 nm yielded the highest diagnostic accuracy for lesions on both occlusal and proximal surfaces while radiography yielded the lowest number of FPs. CONCLUSION: This study demonstrates that simultaneous acquisition of both reflectance and transillumination SWIR images is possible with a single clinical device.

In vivo spectral guided removal of composite from tooth surfaces with a CO2 laser.

Dental composites are used as restorative materials to replace tooth structure after the removal of caries, shaping, covering teeth for esthetic purposes and as adhesives. Dentists spend more time replacing existing restorations that fail than they do placing new restorations. Tooth colored restorations are difficult to differentiate from the surrounding tooth structure making them challenging to remove completely without incidental removal of healthy tooth structure. Previous studies have demonstrated that CO2 lasers in conjunction with spectral feedback can be used to selectively remove composite from tooth surfaces. In addition, we assembled a system feasible for clinical use that incorporates a spectral feedback system, scanning system, articulating arm and a clinical handpiece and subsequently evaluated the performance of that system on extracted teeth. The purpose of this study was to test this system in vivo to demonstrate its efficacy relative to dental clinicians. Eight test subjects with premolar teeth scheduled for extraction for orthodontic reasons had bilateral premolars prepared with small occlusal cavity preparations and filled with dental composite. The laser scanning system was used to remove the composite from one of the preparations and a dental handpiece was used to remove the composite from the other. Cross polarization optical coherence tomography was used to measure the volume of the preparation before and after composite placement and removal. There was no significant difference in the loss of enamel and residual composite between the laser and the handpiece. This study demonstrated that a computer controlled spectral guided CO2 laser scanning system can be used in vivo to selectively remove composite from tooth surfaces.

SWIR, Thermal and CP-OCT imaging probes for the in vivo assessment of the activity of root caries lesions.

New imaging technologies are needed for the clinical assessment of lesions on root surfaces. It is not sufficient to simply detect caries lesions; methods are needed to assess lesion depth, structural composition and activity to determine if chemical intervention has the potential to be effective and if remineralization has occurred. Lesions were monitored using CP-OCT during lesion dehydration to assess the lesion structure and any shrinkage. Thermal imaging at 6-10 µm wavelengths and short wavelength-IR imaging at 1450-1750-nm were used to monitor thermal emission during lesion dehydration to assess lesion activity. Imaging probes were custom fabricated for clinical use. We present the first clinical results of a small feasibility study employing CP-OCT, thermal and SWIR imaging to assess lesion activity in vivo on thirty test subjects with suspected root caries lesions.

Response of Liver Metabolic Pathways to Ketogenic Diet and Exercise Are Not Additive.

PURPOSE: Studies suggest ketogenic diets (KD) produce favorable outcomes (health and exercise performance); however, most rodent studies have used a low-protein KD, which does not reflect the normal- to high-protein KD used by humans. Liver has an important role in ketoadaptation due to its involvement in gluconeogenesis and ketogenesis. This study was designed to test the hypothesis that exercise training (ExTr) while consuming a normal-protein KD (NPKD) would induce additive/synergistic responses in liver metabolic pathways. METHODS: Lean, healthy male C57BL/6J mice were fed a low-fat control diet (15.9% kcal protein, 11.9% kcal fat, 72.2% kcal carbohydrate) or carbohydrate-deficient NPKD (16.1% protein, 83.9% kcal fat) for 6 wk. After 3 wk on the diet, half were subjected to 3-wk treadmill ExTr (5 d·wk, 60 min·d, moderate-vigorous intensity). Upon conclusion, metabolic and endocrine outcomes related to substrate metabolism were tested in liver and pancreas. RESULTS: NPKD-fed mice had higher circulating ß-hydroxybutyrate and maintained glucose at rest and during exercise. Liver of NPKD-fed mice had lower pyruvate utilization and greater ketogenic potential as evidenced by higher oxidative rates to catabolize lipids (mitochondrial and peroxisomal) and ketogenic amino acids (leucine). ExTr had higher expression of the gluconeogenic gene, Pck1, but lower hepatic glycogen, pyruvate oxidation, incomplete fat oxidation, and total pancreas area. Interaction effects between the NPKD and ExTr were observed for intrahepatic triglycerides, as well as genes involved in gluconeogenesis, ketogenesis, mitochondrial fat oxidation, and peroxisomal markers; however, none were additive/synergistic. Rather, in each instance the interaction effects showed the NPKD and ExTr opposed each other. CONCLUSIONS: An NPKD and an ExTr independently induce shifts in hepatic metabolic pathways, but changes do not seem to be additive/synergistic in healthy mice.

Low-intensity exercise induces acute shifts in liver and skeletal muscle substrate metabolism but not chronic adaptations in tissue oxidative capacity.

Adaptations in hepatic and skeletal muscle substrate metabolism following acute and chronic (6 wk; 5 days/wk; 1 h/day) low-intensity treadmill exercise were tested in healthy male C57BL/6J mice. Low-intensity exercise maximizes lipid utilization; therefore, we hypothesized pathways involved in lipid metabolism would be most robustly affected. Acute exercise nearly depleted liver glycogen immediately postexercise (0 h), whereas hepatic triglyceride (TAG) stores increased in the early stages after exercise (0-3 h). Also, hepatic peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) gene expression and fat oxidation (mitochondrial and peroxisomal) increased immediately postexercise (0 h), whereas carbohydrate and amino acid oxidation in liver peaked 24-48 h later. Alternatively, skeletal muscle exhibited a less robust response to acute exercise as stored substrates (glycogen and TAG) remained unchanged, induction of PGC-1α gene expression was delayed (up at 3 h), and mitochondrial substrate oxidation pathways (carbohydrate, amino acid, and lipid) were largely unaltered. Peroxisomal lipid oxidation exhibited the most dynamic changes in skeletal muscle substrate metabolism after acute exercise; however, this response was also delayed (peaked 3-24 h postexercise), and expression of peroxisomal genes remained unaffected. Interestingly, 6 wk of training at a similar intensity limited weight gain, increased muscle glycogen, and reduced TAG accrual in liver and muscle; however, substrate oxidation pathways remained unaltered in both tissues. Collectively, these results suggest changes in substrate metabolism induced by an acute low-intensity exercise bout in healthy mice are more rapid and robust in liver than in skeletal muscle; however, training at a similar intensity for 6 wk is insufficient to induce remodeling of substrate metabolism pathways in either tissue. NEW & NOTEWORTHY Effects of low-intensity exercise on substrate metabolism pathways were tested in liver and skeletal muscle of healthy mice. This is the first study to describe exercise-induced adaptations in peroxisomal lipid metabolism and also reports comprehensive adaptations in mitochondrial substrate metabolism pathways (carbohydrate, lipid, and amino acid). Acute low-intensity exercise induced shifts in mitochondrial and peroxisomal metabolism in both tissues, but training at this intensity did not induce adaptive remodeling of metabolic pathways in healthy mice.

Near-infrared imaging of demineralization on the occlusal surfaces of teeth without the interference of stains.

Most new caries lesions are found in the pits and fissures of the occlusal surface. Radiographs have extremely low sensitivity for early occlusal decay, and by the time the lesion is severe enough to appear on a radiograph, it typically has penetrated well into the dentin and surgical intervention is required. The occlusal surfaces are often heavily stained, and visual and tactile detection have poor sensitivity and specificity. Previous near-infrared imaging studies at wavelengths beyond 1300 nm have demonstrated that stains are not visible and demineralization on the occlusal surfaces can be viewed without interference from stains. The objective of our study is to determine how the contrast between sound and lesion areas on occlusal surfaces varies with wavelength from the visible to 2350 nm and determine to what degree stains interfere with that contrast. The lesion contrast for reflectance is measured in 55 extracted teeth with suspected occlusal lesions from 400 to 2350 nm employing silicon and indium gallium arsenide imaging arrays. In addition, the lesion contrast is measured on 25 extracted teeth with suspected occlusal lesions from 400 to 1600 nm in reflectance and from 830 to 1400 nm in transillumination before and after stains are removed using a ultrasonic scaler. The highest lesion contrast in reflectance is measured at wavelengths >1700 nm. Stains interfere significantly at wavelengths <1150 nm (400 to 1150) for both reflectance and transillumination measurements. Our study suggests that the optimum wavelengths for imaging decay in the occlusal surfaces are >1700 nm for reflectance (1700 to 2350 nm) and near 1300 nm (1250 to 1350 nm) for transillumination.

Probing the brain with molecular fMRI.

One of the greatest challenges of modern neuroscience is to incorporate our growing knowledge of molecular and cellular-scale physiology into integrated, organismic-scale models of brain function in behavior and cognition. Molecular-level functional magnetic resonance imaging (molecular fMRI) is a new technology that can help bridge these scales by mapping defined microscopic phenomena over large, optically inaccessible regions of the living brain. In this review, we explain how MRI-detectable imaging probes can be used to sensitize noninvasive imaging to mechanistically significant components of neural processing. We discuss how a combination of innovative probe design, advanced imaging methods, and strategies for brain delivery can make molecular fMRI an increasingly successful approach for spatiotemporally resolved studies of diverse neural phenomena, perhaps eventually in people.

Multispectral near-infrared reflectance and transillumination imaging of occlusal carious lesions: Variation in lesion contrast with lesion depth.

In vivo and in vitro studies have demonstrated that near-infrared (NIR) light at λ=1300-1700-nm can be used to acquire high contrast images of enamel demineralization without interference of stains. The objective of this study was to determine if a relationship exists between the NIR image contrast of occlusal lesions and the depth of the lesion. Extracted teeth with varying amounts of natural occlusal decay were measured using a multispectral-multimodal NIR imaging system which captures λ=1300-nm occlusal transillumination, and λ=1500-1700-nm cross-polarized reflectance images. Image analysis software was used to calculate the lesion contrast detected in both images from matched positions of each imaging modality. Samples were serially sectioned across the lesion with a precision saw, and polarized light microscopy was used to measure the respective lesion depth relative to the dentinoenamel junction. Lesion contrast measured from NIR cross-polarized reflectance images positively correlated (p<0.05) with increasing lesion depth and a statistically significant difference between inner enamel and dentin lesions was observed. The lateral width of pit and fissures lesions measured in both NIR cross-polarized reflectance and NIR transillumination positively correlated with lesion depth.

Image-guided Removal of Interproximal Lesions with a CO2 Laser.

Recent studies have shown that near-IR (NIR) imaging methods such as NIR reflectance can be used to image lesions on proximal surfaces, and optical coherence tomography (OCT) can be used to measure the depth of those lesions below the tooth surface. These imaging modalities can be used to acquire high contrast images of demineralized tooth surfaces, and 2-D and 3-D images can be extracted from this data. At NIR wavelengths longer than 1200-nm, there is no interference from stains and the contrast is only due to the increased light scattering of the demineralization. Previous studies have shown that image-guided laser ablation can be used to remove occlusal lesions, but its use for the removal of subsurface lesions on proximal surfaces has not been investigated. The objective of this study is to demonstrate that simultaneously scanned NIR and CO2 lasers can be used to selectively remove natural and artificial interproximal caries lesions with minimal damage to sound tooth structure. In this study, images of simulated and natural interproximal lesions on extracted teeth were imaged using a digital microscope, a scanned 1460-nm superluminescent laser diode with an InGaAs detector and a cross polarization OCT system operating at 1300-nm. The lesions were subsequently removed with a CO2 laser operating at 9.3-µm and the dental handpiece and the volume of sound tissue removed was compared.

SWIR reflectance imaging of demineralization on the occlusal surfaces of teeth beyond 1700 nm.

Most new lesions are found in the pits and fissures of the occlusal surface. Radiographs have extremely low sensitivity for early occlusal decay and by the time the lesion is severe enough on a radiograph it typically has penetrated well into the dentin and surgical intervention is required. The occlusal surfaces are heavily stained and visual and tactile methods for their detection also have poor sensitivity and specificity. Previous studies at wavelengths beyond 1300-nm have demonstrated that stains are not visible and demineralization on the occlusal surfaces can be viewed without interference from stains. New extended range InGaAs near-IR cameras allow access to wavelengths beyond 1700-nm. The objective of this study was to determine how the contrast of occlusal lesions varies with wavelength from the visible to 2350-nm. The lesion contrast was measured in 55 extracted teeth with suspected occlusal lesions using reflectance measurements from 400-2350-nm using Si and InGaAs imaging arrays. The highest lesion contrast in reflectance was measured at wavelengths greater than 1700-nm. Stains interfered significantly at wavelengths shorter than 1150-nm. This study indicates that the optimum wavelengths for reflectance imaging decay in the occlusal surfaces are greater than 1700-nm.