Laser Speckle Contrast Imaging Research Articles

Dose-related effect of acetylcholine on human gingival blood flow

Abstract Background This study investigates the dose-response relationship of acetylcholine (ACh) on healthy human gingival blood flow (GBF). Understanding this dose-response relationship contributes to studying vasodilatory mechanisms in various pathological conditions. Methods The study involved 22 young healthy men (21 - 32 years) to investigate the dose-response relationship of ACh on GBF. Semi-circular wells were created on the labial surface of the upper right second incisor (FDI #12) and upper left first incisor (FDI #21), including the gingival sulcus, for the application of drugs. ACh-chloride solutions at 0.1, 1, and 10 mg/mL were administered to the gingival sulcus of tooth FDI #12 with a Hamilton syringe. Physiological saline was applied on the contralateral side to FDI #21 as a control. The GBF was measured non-invasively by the laser speckle contrast imaging method in four 1mm high adjacent regions: coronal, midway1, midway2, and apical, and was expressed in a laser speckle perfusion unit (LSPU). After the baseline blood flow recording, ACh doses were applied sequentially, with washout periods in between. Data were statistically analyzed using a linear mixed model. Results The GBF did not change on the saline site throughout the experiment. The GBF was significantly higher at the coronal region after all ACh doses (baseline: 218±31 LSPU, and 227±38 LSPU p < 0.05, 239±40 LSPU p < 0.001, 291±54 LSPU p < 0.001, respectively) compared to the saline. It was also elevated following 1 and 10 mg/mL at the midway1 (245±48 LSPU, p < 0.05, 293±65 LSPU p < 0.001). At midway2 and apical, only the 10 mg/mL dose was effective (285±71 LSPU, p < 0.001; 302±82 LSPU, p < 0.001). Conclusions Our findings suggest a dose-dependent vasodilation to ACh, emphasizing its role in human gingival microcirculation. Only the 10 mg/mL ACh could evoke remote vasodilation 3 mm from the application. The described method could facilitate the investigation of endothelium-dependent vasodilation in disorders affecting microcirculation, such as periodontitis or diabetes.

Pre-vascularisation of the acellular bladder matrix using the omentum in a porcine model prior for bladder reconstruction-a step towards clinical application?

Research studies with porcine acellular bladder matrix (PABM) showed integration of only small sized stamps in recipient bladders, however for clinical use in bladder augmentation significantly larger patches are needed. We hypothesised pre-vascularisation with omentum may be a step towards clinical translation. Eight domestic pigs were operated three times 8-10weeks apart: 1-Implantation; PABM with recorded dimensions were sutured around a tissue expanding device, wrapped in omentum and sutured to the anterior abdominal wall. 2-Augmentation; hemi-cystectomy and bladder augmentation was performed with the pre-vascularized PABM using non-absorbable suture 3-Sacrifice; The dimensions of the PABMs were measured macroscopically, the in-vivo microcirculation of the PABMs were assessed using laser speckle contrast imaging. HE staining, uroplakin 3 and CK7 immunohistochemistry was performed. In seven animals, the bladder augmentation was successful without complication. One animal was lost in bowel obstruction and in two animals enteric fistula was found after the first intervention. The rectangular shape of the initial tissue expander was subsequently changed. All the seven patches were strong, compliant and had integrated with the surrounding native bladder and were 83% (48-100%) of the original patch size. Laser speckle contrast imaging already showed microcirculation at the patch edges at augmentation and this further improved towards the centre of the patches by the end of the study. Histology demonstrated acute inflammatory response with fibroblast infiltration after implantation and full coverage by urothelium was seen with positive staining for CK7 antibodies. Pre-vascularization of PABM in the omentum of healthy porcine models allows larger PABM patches to integrate this may be a step towards clinical application.

Quantum machine learning enhanced laser speckle analysis for precise speed prediction.

Laser speckle contrast imaging (LSCI) is an optical technique used to assess blood flow perfusion by modeling changes in speckle intensity, but it is generally limited to qualitative analysis due to difficulties in absolute quantification. Three-dimensional convolutional neural networks (3D CNNs) enhance the quantitative performance of LSCI by excelling at extracting spatiotemporal features from speckle data. However, excessive downsampling techniques can lead to significant information loss. To address this, we propose a hybrid quantum-classical 3D CNN framework that leverages variational quantum algorithms (VQAs) to enhance the performance of classical models. The proposed framework employs variational quantum circuits (VQCs) to replace the 3D global pooling layer, enabling the model to utilize the complete 3D information extracted by the convolutional layers for feature integration, thereby enhancing velocity prediction performance. We perform cross-validation on experimental LSCI speckle data and demonstrate the superiority of the hybrid models over their classical counterparts in terms of prediction accuracy and learning stability. Furthermore, we evaluate the models on an unseen test set and observe that the hybrid models outperform the classical models with up to 14.8% improvement in mean squared error (MSE) and up to 26.1% improvement in mean absolute percentage error (MAPE) evaluation metrics. Finally, our qualitative analysis shows that the hybrid models offer substantial improvements over classical models in predicting blood flow at both low and high velocities. These results indicate that the hybrid models possess more powerful learning and generalization capabilities.

Pressure-Induced Microvascular Reactivity With Whole Foot Loading Is Unique Across the Human Foot Sole.

Foot sole plantar pressure generates transient but habitual cutaneous ischemia, which is even more exacerbated in atypical gait patterns. Thus, adequate post-occlusive reactive hyperaemia (PORH) is necessary to maintain skin health. Plantar pressure regional variance during daily tasks potentially generates region-specific PORH, crucial for ischemic defence. The current work investigated regional PORH across the human foot sole resulting from stance-like loading. A loading device equipped with an in-line laser speckle contrast imager measured blood flux before, during, and after whole-foot loading for 2 and 10 min durations at 15% and 50% body weight. Flux was compared between six regions: the heel, lateral arch, medial arch, and fifth, third, and first metatarsals (MT). Baseline flux was significantly greater in the 1MT and 3MT than all other regions. Loading occluded the heel, 5MT and 3MT more than all other regions. Regional PORH peak, time to peak, area under the curve, and recovery rate were ranked between regions. The 3MT, followed by 5MT, overall had the strongest PORH response, suggesting a heightened protection against ischemia compared to other regions. This work highlights regional variations within a healthy foot, providing a framework for future ulcer risk assessments and interventions to preserve foot health.

Factors Associated With Ocular Perfusion Measurements as Obtained With Laser Speckle Contrast Imaging.

To study the ocular and systemic factors affecting optic nerve head (ONH) perfusion data as obtained using a commercially available laser speckle flowgraphy (LSFG) device in a cohort of Caucasian subjects without ocular diseases. Also, to assess the intrasession repeatability and intersession reproducibility of ONH, macular, retinal, and choroidal perfusion. Seventy-five healthy eyes of 75 Caucasian participants underwent LSFG and spectral-domain optical coherence tomography (SD-OCT) on the same visit. Perfusion of the ONH was assessed with LSFG, and SD-OCT was used to measure peripapillary retinal nerve fiber layer thickness (RNFLT) and macular ganglion cell plus inner plexiform layer thickness (GCIPLT). The intrasession repeatability and intersession reproducibility of ONH and macular perfusion and retinal and choroidal relative flow volume (RFV) were evaluated in 20 participants measured on three different days over a 6-month period. Intrasession and intersession intraclass correlation coefficients of LSFG parameters ranged from 0.787 to 0.967 and from 0.776 to 0.935, respectively. Intersession 95% prediction intervals for the ratio of two measurements were wider for RFV indices than for ONH and macular perfusion parameters. The multiple regression analysis indicated that higher ONH perfusion was associated with younger age, female sex, smaller optic disc area, and higher RNFLT. RNFLT was an independent predictor of ONH perfusion, whereas GCIPLT was not. Each 1-µm increase in RNFLT was associated with a 0.272 arbitrary unit increase in ONH perfusion. LSFG measurements of optic disc perfusion are influenced by sex, age, and anatomical variations in optic disc area and RNFLT. Better evaluation of ocular blood flow will result in better diagnosis and treatment of various ocular diseases.

Study on the Correlation Between Renal Blood Perfusion and Kidney Injury in Different Weekly-Aged Type 2 Diabetic Mice

This study aims to explore the correlation between renal blood perfusion (RBP) and diabetic nephropathy (DN). Methods: A total of 72 mice included db/db and db/m mice at the ages of 6, 14, and 22 weeks, forming six groups. RBP was assessed using Laser Speckle Contrast Imaging (LSCI). Kidney function markers and the extent of pathological damage were evaluated. Pearson correlation analysis was employed to predict the relationship between RBP and various indicators of kidney damage. Results: Compared to db/m mice of all ages, 6-week-old db/db mice showed no significant difference in kidney function markers and had no apparent pathological damage. However, db/db mice at other ages showed deteriorating kidney functions and evident pathological damage, which worsened with age. The RBP in db/m mice of all ages and 6-week-old db/db mice showed no significant difference; however, RBP in db/db mice demonstrated a significant declining trend with age. The correlation between RBP and kidney damage indicators was as follows: 24 h urinary microalbumin (r=-0.728), urinary transferrin (r=-0.834), urinary β2-microglobulin (r=-0.755), urinary monocyte chemoattractant protein-1 (r=-0.786), Masson's trichrome staining (r=-0.872), and Periodic Acid-Schiff staining (r=-0.908). Conclusion: RBP is strongly correlated with the extent of diabetic kidney damage.

Cerebral Cortical Vasodilation via Nicotinic Receptors by Heated Tobacco Product Aerosol Extract in Rats

Introduction: Smoking increases the risk of lung cancer due to a number of components of smoke. The use of novel heated tobacco products (HTPs), alternative to conventional combustion cigarettes, has increased in recent years. However, the in vivo biological effects of HTPs are poorly understood. This study aimed to clarify the acute effects of injecting aerosol extract prepared from an HTP on regional cerebral blood flow (rCBF) in rat cortex by comparing them to the effects of injecting smoke extract prepared from conventional combustible cigarettes. Methods: In urethane anesthetized rats, rCBF was measured using laser speckle contrast imaging simultaneously with arterial pressure. Results: Both cigarette smoke extract and HTP aerosol extract, at a dose equivalent to 30 μg nicotine/kg, injected intravenously, increased cortical rCBF without changing arterial pressure. The magnitude and time course of the increased rCBF response to both extracts were similar throughout the cortical area, and the rCBF increases were all abolished by dihydro-β-erythroidine, an α4β2-preferring nicotinic acetylcholine receptor (nAChR) antagonist. Conclusion: In conclusion, our study demonstrated that the effect of injecting aerosol extract prepared from an HTP, an acute increase in cortical rCBF, is mediated via activation of α4β2-like neuronal nAChRs in the brain.

Impact of light polarization on laser speckle contrast imaging with a custom phantom for microvascular flow

Laser speckle contrast imaging (LSCI) is a non-invasive, powerful, and cost-effective imaging technique that has seen widespread adoption across various medical fields, particularly for blood flow imaging. While LSCI provides physicians with valuable insights into changes or occlusions in blood flow, the technique is susceptible to various factors and parameters that can impact measurement sensitivity and signal-to-noise ratio (SNR). These include the scattering of light, which can affect the quality and reliability of the LSCI data. The polarization of light holds significant promise to enhance the performance of LSCI. In this study, we employed polarization manipulation of light to investigate its impact on the performance of LSCI for measuring flow. Focusing on the application of LSCI in microcirculation within capillaries, we examined the effect of polarization control on the technique's flow measurement capabilities using a custom-designed phantom system. This phantom consisted of three tubes with inner diameters of 1.1 mm, 1.6 mm, and 2.8 mm, embedded in a polydimethylsiloxane (PDMS) matrix with optical properties similar to biological tissue. By manipulating the polarization of both the incident and reflected light, alternating between parallel and perpendicular states, we compared the performance of our LSCI system in detecting flow for different tube diameters and depths within the phantom. Our study revealed that while depth is a critical parameter influencing flow detection using LSCI, employing perpendicular polarization (between incident and reflected light) resulted in the lowest measurement error and highest SNR compared to parallel polarization and the absence of polarization control.

Cerebral blood flow patterns induced by photoactivation based on laser speckle contrast imaging

Cerebral blood flow patterns induced by photoactivation based on laser speckle contrast imaging

An integrated portable system for laser speckle contrast imaging and digital holographic microscopy

A multimodal quantitative phase imaging platform combining digital holographic microscopy (DHM) with laser speckle contrast imaging (LSCI) is demonstrated for imaging weakly scattering and transparent samples in a label-free manner. It uses the principle of coherence of the light source for interferometric detection of phase of transparent and semi-transparent samples. The speckle formation resulting from the coherence property of the laser source is used to track dynamic activities in the regions of interest in the sample. Integration of these two techniques onto a microfluidic chip leads to an optofluidic real-time microscope for live cell imaging applications. In this work, we have developed an integrated multimodal system combining digital holographic microscopy and laser speckle contrast imaging system for Optofluidics and in vitro studies. The sample flowing through the microfluidic channel is imaged to record holograms and video of intensity images at the same time. This enables to map the flow within a microfluidic channel and quantify the channel as well as the particle flow through the channel. The channel morphology along with the particles flowing through the channel are quantified using DHM. The two-dimensional speckle contrast images map the flow of the dynamic microbeads and cells with very high contrast in almost real time. A low-cost portable multimodal quantitative phase microscope combining DHM and LSCI has been demonstrated for real time imaging with applications in optofluidics.

Correlation between maternal microvascular reactivity assessed by laser speckle contrast imaging and first trimester placental biomarkers

Correlation between maternal microvascular reactivity assessed by laser speckle contrast imaging and first trimester placental biomarkers

Skin microvascular function assessed by laser speckle contrast imaging in twin pregnancies

Skin microvascular function assessed by laser speckle contrast imaging in twin pregnancies

Evaluation of maternal skin microvascular reactivity by laser speckle contrast imaging in pregnancies complicated by preeclampsia and foetal growth restriction

Evaluation of maternal skin microvascular reactivity by laser speckle contrast imaging in pregnancies complicated by preeclampsia and foetal growth restriction

Improving the early diagnosis and clinical outcomes of shock patients via laser speckle contrast imaging assessment of peripheral hemodynamics

Improving the early diagnosis and clinical outcomes of shock patients via laser speckle contrast imaging assessment of peripheral hemodynamics

Comprehensive validation of a compact laser speckle contrast imaging system for vascular function assessment: from the laboratory to the clinic.

Proper organ functioning relies on adequate blood circulation; thus, monitoring blood flow is crucial for early disease diagnosis. Laser speckle contrast imaging (LSCI) is a noninvasive technique that is widely used for measuring superficial blood flow. In this study, we developed a portable LSCI system using an 805-nm near-infrared laser and a monochrome CMOS camera with a 10 × macro zoom lens. The system achieved a high-resolution imaging (1280 × 1024 pixels) with a working distance of 10 to 35 cm. The relative flow velocities were visualized via a spatial speckle contrast analysis algorithm with a 5 × 5 sliding window. In vitro experiments demonstrated the system's ability to image flow velocities in a fluid model, and a linear relationship was observed between the actual flow rate and the relative flow rate obtained by the system. The correlation coefficient (R2) exceeded 0.83 for volumetric flow rates of 0 to 0.2 ml/min when channel widths were greater than 1.2 mm, and R2 > 0.94 was obtained for channel widths exceeding 1.6 mm. Comparisons with laser Doppler flowmetry (LDF) revealed a strong positive correlation between the LSCI and LDF results. In vivo experiments captured postocclusive reactive hyperemic responses in rat hind limbs and human palms and feet. The main research contribution is the development of this compact and portable LSCI device, as well as the validation of its reliability and convenience in various scenarios and environments. Future applications of this technology include evaluating blood flow changes during skin injuries, such as abrasions, burns, and diabetic foot ulcers, to aid medical institutions in treatment optimization and to reduce treatment duration.

H2S-Eluting Hydrogels Promote In Vitro Angiogenesis and Augment In Vivo Ischemic Wound Revascularization

Ischemic wounds are frequently encountered in clinical practice and may be related to ischemia secondary to diabetes, peripheral artery disease and other chronic conditions. Angiogenesis is critical to the resolution of ischemia. Hydrogen sulfide (H2S) is now recognized as an important factor in this process. H2S donors NaHS and GYY4137 were incorporated into the photosensitive polymer hydrogel gelatin methacrylate and evaluated. Human umbilical vein endothelial cell (HUVEC) culture was used to quantify toxicity and angiogenesis. Sprague Dawley rats were subjected to ischemic myocutaneous flap wound creation with and without application of H2S-eluting hydrogels. Tissue perfusion during wound healing was quantified using laser speckle contrast imaging, and gene and protein expression for VEGF were evaluated. Vascular density was assessed by CD31 immunohistochemistry. Successful incorporation of sulfide compounds was confirmed by scanning electron microscopy with energy-dispersive X-ray analysis, and under physiologic conditions, detectable H2S was present for up to 14 days by high-performance liquid chromatography. HUVECs exposed to hydrogels did not demonstrate excess cytotoxicity or apoptosis. A two-fold increase in angiogenic tube formation was observed in HUVECs exposed to H2S-eluting hydrogels. Rat ischemic flap wounds demonstrated greater perfusion at 14 days, and there was greater vascularity of healed wounds compared to untreated animals. A nearly two-fold increase in VEGF mRNA and a four-fold increase in VEGF protein expression were present in wounds from treated animals. Local-regional administration of H2S represents a novel potential therapeutic strategy to promote angiogenesis and improve wound healing after tissue injury or as a result of ischemic disease.

Increased Levels of Circulating Methylglyoxal Have No Consequence for Cerebral Microvascular Integrity and Cognitive Function in Young Healthy Mice.

Diabetes and other age-related diseases are associated with an increased risk of cognitive impairment, but the underlying mechanisms remain poorly understood. Methylglyoxal (MGO), a by-product of glycolysis and a major precursor in the formation of advanced glycation end-products (AGEs), is increased in individuals with diabetes and other age-related diseases and is associated with microvascular dysfunction. We now investigated whether increased levels of circulating MGO can lead to cerebral microvascular dysfunction, blood-brain barrier (BBB) dysfunction, and cognitive impairment. Mice were supplemented or not with 50mM MGO in drinking water for 13weeks. Plasma and cortical MGO and MGO-derived AGEs were measured with UPLC-MS/MS. Peripheral and cerebral microvascular integrity and inflammation were investigated. Cerebral blood flow and neurovascular coupling were investigated with laser speckle contrast imaging, and cognitive tests were performed. We found a 2-fold increase in plasma MGO and an increase in MGO-derived AGEs in plasma and cortex. Increased plasma MGO did not lead to cerebral microvascular dysfunction, inflammation, or cognitive decline. This study shows that increased concentrations of plasma MGO are not associated with cerebral microvascular dysfunction and cognitive impairment in healthy mice. Future research should focus on the role of endogenously formed MGO in cognitive impairment.

Images in Vascular Medicine: Analysis of skin microvascular dysfunction in Harlequin syndrome using laser speckle contrast imaging.

Images in Vascular Medicine: Analysis of skin microvascular dysfunction in Harlequin syndrome using laser speckle contrast imaging.

Persistent Na + current couples spreading depolarization to seizures in Scn8a gain of function mice.

Spreading depolarization (SD) is a slowly propagating wave of massive cellular depolarization that transiently impairs the function of affected brain regions. While SD typically arises as an isolated hemispheric event, we previously reported that reducing M-type potassium current (I KM ) by ablation of Kcnq2 in forebrain excitatory neurons results in tightly coupled spontaneous bilateral seizure-SD complexes in the awake mouse cortex. Here we find that enhanced persistent Na + current due to gain-of-function (GOF) mutations in Scn8a (N1768D/+, hereafter D/+) produces a similar compound cortical excitability phenotype. Chronic DC-band EEG recording detected spontaneous bilateral seizure-SD complexes accompanied by seizures with a profound tonic component, which occurs predominantly during the light phase and were detected in the mutant mice across ages between P40-100. Laser speckle contrast imaging of cerebral blood flow dynamics resolved SD as bilateral wave of hypoperfusion and subsequent hour-lasting hypoperfusion in Scn8a D/+ cortex in awake head-restrained mice subjected to a subconvulsive PTZ. Subcortical recordings in freely moving mice revealed that approximately half of the spontaneous cortical seizure-SD complexes arose with concurrent SD-like depolarization in the thalamus and delayed depolarization in the striatum. In contrast, SD-like DC potential shifts were rarely detected in the hippocampus or upper pons. Consistent with the high spontaneous incidence in vivo , cortical slices from Scn8a D/+ mice showed a raised SD susceptibility, and pharmacological inhibition of persistent Na + current (I NaP ), which is enhanced in Scn8a D/+ neurons, inhibited SD generation in cortical slices ex vivo , indicating that I NaP contributes to SD susceptibility. Ex vivo Ca 2+ imaging studies using acute brain slices expressing genetic Ca 2+ sensor (Thy1-GCAMP6s) demonstrated that pharmacological activation of I KM suppressed Ca 2+ spikes and SD, whereas I KM inhibitor drastically increased the frequency of Ca 2+ spikes in the hippocampus of Scn8a D/+ mice, but not in WT, suggesting that I KM restrains the hyperexcitability resulting from Scn8a GOF mutation. Together, our study identifies a cortical SD phenotype in Scn8a GOF mice shared with the Kcnq2 - cKO model of developmental epileptic encephalopathy and reveals that an imbalance of non-inactivating inward and outward membrane currents bidirectionally modulates spatiotemporal SD susceptibility.

Statistical assessment of the inner shape of a channel based on laser speckle contrast imaging

In this work, we employed laser speckle contrast imaging coupled with the analysis of several statistical moments on the recorded images to recognize the shape of an object through the analysis of the pattern of a flow around it. We postulated that a flow through different geometries would generate a unique flow structure and thus the mathematical analysis of the differences can be utilized to detect and recognize various shape geometries. To this aim, the flow of scattered liquid that had passed through objects of different shapes was illuminated by a laser beam and the diffused speckle image was recorded by a camera. In the computer, the captured speckle image was first converted to a flow image and then mathematically analyzed to recognize the shape of the objects. Three out of the eight statistical metrics were found to be the best candidates for the recognition of shapes, thus proving our hypothesis.