In order to enhance the optical clearing effect of topically applied optical clearing agents(OCAs),we evaluated the effect of propylene glycol(PG)as a chemical penetration enhancer(PE)on optical clearing of skin in vi...In order to enhance the optical clearing effect of topically applied optical clearing agents(OCAs),we evaluated the effect of propylene glycol(PG)as a chemical penetration enhancer(PE)on optical clearing of skin in vitro by observation and measurement of optical-transmittance and diffuse-reflectance spectra.Three OCAs,i.e.,glycerol,D-sorbitol and PEG400,and two other penetration enhancers,Azone and Thiazone,were used in this study.The results indicated that the decrease of reduced scattering coefficient caused by OCA/PG was larger than that by pure OCA,and the change by OCA/water was the least after the same treatment time.There were significant differences for the reduced scattering coefficient at 630 nm after 120 min application of agents between OCA and OCA/PG.The efficacy of optical clearing caused by OCA/PG depended on the OCA itself.When PEG400 was mixed with three different PEs,we found the optical clearing were different.The penetration enhancing ability of PG was much better compared to Azone,and suboptimal to Thiazone.Also,this study provides evidence for the use of PG as a PE in order to improve skin optical clearing.展开更多
Confocal Raman microspectroscopy(CRM)with 633-and 785-nm excitation wavelengths combined with optical clearing(OC)technique was used for ex-vivo study of porcine skin in the Raman fingerprint region.The optical cleari...Confocal Raman microspectroscopy(CRM)with 633-and 785-nm excitation wavelengths combined with optical clearing(OC)technique was used for ex-vivo study of porcine skin in the Raman fingerprint region.The optical clearing has been performed on the skin samples by applying a mixture of glycerol and distilled water and a mixture of glycerol,distilled water and chemical penetration enhancer dimethyl sulfoxide(DMSO)during 30 min and 60 min of treat-ment.It was shown that the combined use of the optical clearing technique and CRM at 633 nm allowed one to preserve the high probing depth,signal-to-noise ratio and spectral resolution simultaneously.Comparing the effect of different optical clearing agents on porcine skin showed that an optical clearing agent containing chemical penetration enhancer provides higher optical clearing efficiency.Also,an increase in treatment time allows to improve the optical clearing efficiency of both optical clearing agents.As a result of optical clearing,the detection of the amide-Ⅲ spectral region indicating well-distinguishable structural differences between the type-Ⅰ and type-Ⅳ collagens has been improved.展开更多
Our previous studies demonstrated the ultrasound-induced skin optical clearing enhancement with topical application of optical clearing agents on in vitro porcine skin and in vivo human skin.The objective of this stud...Our previous studies demonstrated the ultrasound-induced skin optical clearing enhancement with topical application of optical clearing agents on in vitro porcine skin and in vivo human skin.The objective of this study was to investigate the possible mechanisms of the enhanced skin optical clearing by ultrasound medications.Optical clearing effects of ex vivo guinea pig abdomen skin topically applied with 60% glycerol or the combination of 60% glycerol and ultrasound were studied by optical coherence tomography(OCT).Microstructure of skin surface was examined by scanning electron microscopy(SEM).Ultrasound with a frequency of 1MHz and a power of 0.75W over a 3-cm probe was simultaneously applied with glycerol solution for 15min.The combination of 60% glycerol and ultrasound results in a 19% increase in OCT 1/e light penetration depth after 30min,which is much better than 60% glycerol alone.SEM images demonstrated that changes in skin microstructure due to the tight order of the lipid bilayers in the stratum corneum disrupted and the separation of keratinocytes by the application of ultrasound contribute to the ultrasound-enhanced intact skin optical clearing effects.展开更多
The effect of optical cleaning method combined with laser speckle imaging(LSI)was discussed to improve the detection depth of LSI due to high scattering characteristics of skin,which limit its clinical application.A d...The effect of optical cleaning method combined with laser speckle imaging(LSI)was discussed to improve the detection depth of LSI due to high scattering characteristics of skin,which limit its clinical application.A double-layer skin tissue model embedded with a single blood vessel was established,and the Monte Carlo method was used to simulate photon propagation under the action of light-permeating agent.808 nm semiconductor and 632.8 nm He–Ne lasers were selected to study the e®ect of optical clearing agents(OCAs)on photon deposition in tissues.Results show that the photon energy deposition density in the epidermis increases with the amount of tissue°uid replaced by OCA.Compared with glucose solution,polyethylene glycol 400(PEG 400)and glycerol can considerably increase the average penetration depth of photons in the skin tissue,thereby raising the sampling depth of the LSI.After the action of glycerol,PEG 400,and glucose,the average photon penetration depth is increased by 51.78%,51.06%,and 21.51%for 808nm,68.93%,67.94%,and 26.67%for 632.8 nm lasers,respectively.In vivo experiment by dorsal skin chamber proves that glycerol can cause a substantial decrease in blood°ow rate,whereas PEG 400 can signicantly improve the capability of light penetration without a®ecting blood velocity,which exhibits considerable potential in the monitoring of blood°ow in skin tissues.展开更多
Tissue Optical Clearing Devices(TOCDs)have been shown to increase light transmission through mechanically compressed regions of naturally turbid biological tissues.We hypothesize that zones of high compressive strain ...Tissue Optical Clearing Devices(TOCDs)have been shown to increase light transmission through mechanically compressed regions of naturally turbid biological tissues.We hypothesize that zones of high compressive strain induced by TOCD pins produce localized water displacement and reversible changes in tissue optical properties.In this paper,we demonstrate a novel combined mechanical finite element model and optical Monte Carlo model which simulates TOCD pin compression of an ex vivo porcine skin sample and modified spatial photon fluence distributions within the tissue.Results of this simulation qualitatively suggest that light transmission through the skin can be significantly affected by changes in compressed tissue geometry as well as concurrent changes in tissue optical properties.The development of a comprehensive multi-domain model of TOCD application to tissues such as skin could ultimately be used as a framework for optimizing future design of TOCDs.展开更多
Laser Speckle Contrast Imaging(LSCI)plays an important role in studying blood flow,but suffers from limited penetration depth of light in turbid tissue.The strong scattering of tissue obviously reduces the image contr...Laser Speckle Contrast Imaging(LSCI)plays an important role in studying blood flow,but suffers from limited penetration depth of light in turbid tissue.The strong scattering of tissue obviously reduces the image contrast which decreases the sensitivity to flow velocity.Some image processing or optical clearing methods have been proposed to lessen the deficiency,but quantitative assessment of improvement is seldom given.In this study,LSCI was applied to monitor the blood flow through a capillary embedded within various tissue phantoms at depths of 0.25,0.45,0.65,0.85 and 1.05 mm,and the flow velocity in capillary was controllable from 0 to 4mm/s.Here,glycerol,a common optical clearing agent,was mixed with Intralipid at different volume ratio to make the reduced scattering coefficient of tissue phantom decrease from 13.00 to 0.50 cm−1.The quantitative analysis demonstrates that the optical clearing method can obviously enhance the image contrast,imaging depth,and sensitivity to blood flow velocity.Comparing the Laser Speckle Contrast Analysis methods and the optical clearing method,we find that for typical turbid tissue,the sensitivity to velocity estimated by the Laser Speckle Temporal Contrast Analysis(LSTCA)is twice of that by the Laser Speckle Spatial Contrast Analysis(LSSCA);while the sensitivity to velocity estimated by using the two analysis methods has a 10-fold increase,respectively,if addition of glycerol makes the reduced scattering coefficient of tissue phantom decrease by 30%.Combining the LSTCA and the optical clearing method,the sensitivity to flow velocity will be further enhanced.展开更多
Tissue optical clearing by use of optical clearing agents(OCAs)has been proven to have potential to reduce the highly scattering effect of biological tissues in optical techniques.However,the difference in tissue samp...Tissue optical clearing by use of optical clearing agents(OCAs)has been proven to have potential to reduce the highly scattering effect of biological tissues in optical techniques.However,the difference in tissue samples could lead to unreliable results,making it difficult to quantitatively control the dose of OCAs during the course of tissue optical clearing.In this work,in order to study the effects of optical clearing,we customized tissue-like phantoms with optical properties of some biological tissue.Diffuse reflectance and total transmittance of tissue-like phantoms with different OCAs(DMSO or glycerol)and porcine skin tissues were measured.Then optical property parameters were calculated by inverse adding-doubling(IAD)algorithm.Results showed that OCAs could lead to a reduction in scattering of tissue-like phantoms as it did to porcine skin tissue in vitro.Furthermore,a series of relational expressions could be fit to quantitatively describe the relationship between the doses of OCAs and the reduction of scattering effects.Therefore,proper tissue-like phantom could facilitate optical clearing to be used in quantitative control of tissue optical properties,and further promote the application potential of optical clearing to light-based noninvasive diagnostic and therapeutic techniques.展开更多
Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible...Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness.While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D,they often have limitations,such as being too harsh for fragile 3D cell cultures,requiring complex handling protocols,or inducing tissue deformation with shrinkage or expansion.To address this issue,we proposed a modified optical clearing method for 3D cell cultures,called MACS-W,which is simple,highly efficient,and morphology-preserving.In our evaluation of MACS-W,we found that it exhibits excellent clearing capability in just 10 min,with minimal deformation,and helps drug evaluation on tumor spheroids.In summary,MACS-W is a fast,minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures.展开更多
For the first time,the changes in autofluorescence spectra of ex vivo rat skin have been experimentally investigated using the combination of fluorescence spectroscopy and optical immersion clearing.The glucose,glycer...For the first time,the changes in autofluorescence spectra of ex vivo rat skin have been experimentally investigated using the combination of fluorescence spectroscopy and optical immersion clearing.The glucose,glycerol and propylene glycol solutions were used as clearing agents.The optical clearing was performed from the dermal side of skin imitating the in vivo injection of clearing agent under the dermal layers.In this contribution,the common properties of autofluorescence variation during optical immersion clearing were determined.The tendency of autofluorescence signal to decrease with reduction of scattering in tissue was noticed and discussed in detail.However,the differences in the shape of spectral curves under application of different clearing agents showed that optical clearing affects the autofluorescence properties of tissue differently depending on the type of clearing liquid.The results obtained are useful for the understanding of tissue optical clearing mechanisms and for improving techniques such as fluorescence spectroscopy.展开更多
Lasers have been widely used for tattoo removal,but the limited light penetration depth calused by high skin scattering property restricts the therapeutic outcome of deep tattoo.Skin optical clearing method,by introdu...Lasers have been widely used for tattoo removal,but the limited light penetration depth calused by high skin scattering property restricts the therapeutic outcome of deep tattoo.Skin optical clearing method,by introducing optical clearing agent(OCA)into skin,has shown some im-provement in the effect of laser tattoo removal.In this study,the enhanced laser tattoo removal has been quantitatively assessed.OCA was applied to the skin of tattoo animal model and Q switched Nd:YAG laser(1064 nm)irradiation was used to remove the tattoo.The skin evaluation instrument(Mexameter probe,MPA580)was applied to measure the content of tattoo pigment before and after laser treatment,and then the clearance rate of pigment was calculated.Further,Monte Carlo(MC)method was utilized to simulate the efecet of skin optical clearing on light transmission in tattoo skin model.By comparing the pigment change of tattoo areas respectively treated with OCA plus laser and single laser,it was found that pigment clearance of the former tattoo area was increased by 1.5-fold.Further,the MC simulation verifed that the reduced light scattering in skin could increase the effective dose of lumninous fux reaching to the deep tattoo regions.It can be concluded from both experiment and theoretical simulations that skin optical clearing technique could improve the outcome of laser tattoo re moval,which should be beneficial for clinical laser tattoo removal and other laser pigment elimination.展开更多
Vascular-targeted photodynamic therapy(V-PDT)is an effective treatment for port wine stains(PWS).However,repeated treatment is usually needed to achieve optimal treatment outcomes,possibly due to the limited treatment...Vascular-targeted photodynamic therapy(V-PDT)is an effective treatment for port wine stains(PWS).However,repeated treatment is usually needed to achieve optimal treatment outcomes,possibly due to the limited treatment light penetration depth in the PWS lesion.The optical clearing technique can increase light penetration in depth by reducing light scattering.This study aimed to investigate the V-PDT in combination with an optical clearing agent(OCA)for the therapeutic enhancement of V-PDT in the rodent skinfold window chamber model.Vascular responses were closely monitored with laser speckle contrast imaging(LSCI),optical coherence tomography angiography,and stereo microscope before,during,and after the treatment.We further quantitatively demonstrated the effects of V-PDT in combination with OCA on the blood flow and blood vessel size of skin microvasculature.The combination of OCA and V-PDT resulted in significant vascular damage,including vasoconstriction and the reduction of blood flow.Our results indicate the promising potential of OCA for enhancing V-PDT for treating vascular-related diseases,including PWS.展开更多
Objective:Wallerian degeneration is a pathological process closely related to peripheral nerve regeneration following injury,and includes the disintegration and phagocytosis of peripheral nervous system cells.Traditio...Objective:Wallerian degeneration is a pathological process closely related to peripheral nerve regeneration following injury,and includes the disintegration and phagocytosis of peripheral nervous system cells.Traditionally,morphological changes are observed by performing immunofluorescence staining after sectioning,which results in the loss of some histological information.The purpose of this study was to explore a new,nondestmetive,and systematic method for observing axonal histological changes during Wallerian degeneration.Methods:Thirty male Thy1-YFP-16 mice(SPF grade,6 weeks old,20±5 g)were randomly selected and divided into clear,unobstructed brain imaging cocktails and computational analysis(CUBIC)optical clearing(n=15)and traditional method groups(n=15).Five mice in each group were sacrificed at 1st,3rd,and 5th day following a crush operation.The histological axon changes were observed by CUBIC light optical clearing treatment,direct tissue section imaging,and HE staining.Results:The results revealed that,compared with traditional imaging methods,there was no physical damage to the samples,which allowed for three-dimensional and deep-seated tissue imaging through CUBIC.Local image information could be nicely obtained by direct fluorescence imaging and HE staining,but it was difficult to obtain image information of the entire sample.At the same time,the image information obtained by fluorescence imaging and HE staining was partially lost.Conclusion:The combining of CUBIC and Thy1-YFP transgenic mice allowed for a clear and comprehensive observation of histological changes of axons in Wallerian degeneration.展开更多
Organisms are composed of a vast number of highly specialized cells with characteristic physiological functions determined by their structure.Therefore,understanding physiological and pathological mechanisms requires ...Organisms are composed of a vast number of highly specialized cells with characteristic physiological functions determined by their structure.Therefore,understanding physiological and pathological mechanisms requires obtaining 3-dimensional(3D)structural information at the cellular level across tissues,organs,and even the whole body.However,the scattering and absorption of tissue limits the penetration depth of light and thus extremely reduces the imaging performance in deep tissue.It is well known that 3D imaging outperforms 2D imaging in various scenarios for 3D reconstruction of tissue structures.This includes tracing convoluted 3D structures like vasculature,neurons,and prostate glands,as well as elucidating intricate distributions of cells or lesions.展开更多
Complex study of glycerol effects on the skin tissue was performed.The change in optical,weight and geometrical parameters of the rat skin under the action of the glycerol solutions was studied ex vivo.Possible mechan...Complex study of glycerol effects on the skin tissue was performed.The change in optical,weight and geometrical parameters of the rat skin under the action of the glycerol solutions was studied ex vivo.Possible mechanisms of the skin optical clearing under the action of glycerol solutions of different concentrations were discussed.The results can be helpful for refinement of models developed to evaluate the effective diffusion coefficients of glycerol in tissues.展开更多
With the objective to study the variation of optical properties of rat muscle during optical clearing,we have performed a set of optical measurements from that kind of tissue.The.measurements performed were total tr a...With the objective to study the variation of optical properties of rat muscle during optical clearing,we have performed a set of optical measurements from that kind of tissue.The.measurements performed were total tr ansmittance,ollimated transmit tance,specular reflec-tance and total reflectance.This set of measurements is suficient to determine diffuse reflectance and absorbance of the sample,also necessary to est imate the optical properties.All the per formed measurements and calculated quantities will be used later in inverse Monte Carlo(IMC)simu-lations to determine the evolution of the optical properties of muscle during treatments with ethylene glycol and glucose.The results obt ained with the measurements already provide some information about the optical c learing treatments applied to the muscle and translate the mechanisms of turning the tissue more transparent and sequence of regimes of optical clearing.展开更多
以甘油光学清除的小鼠皮肤为研究对象,通过二次谐波(second-harmonic generation,SHG)成像和透射电子显微镜(transmission electron microscope,TEM)研究光学清除对组织超微结构的影响.SHG成像结果表明,甘油处理能够显著增大信号强度并...以甘油光学清除的小鼠皮肤为研究对象,通过二次谐波(second-harmonic generation,SHG)成像和透射电子显微镜(transmission electron microscope,TEM)研究光学清除对组织超微结构的影响.SHG成像结果表明,甘油处理能够显著增大信号强度并提升成像深度,证实其可作为一种有效的光学清除剂.通过TEM对比观察常规化学固定与高压冷冻(high-pressure freezing,HPF)固定样品的超微结构,发现HPF能更好地保持组织自然状态.进一步分析显示,甘油处理虽使胶原纤维直径减小,但其排列周期未发生明显变化,说明光学清除过程未对胶原纤维结构造成损伤.上述结果表明,甘油具有高效、安全的光学清除效果,且在光学清除过程中不会引起组织超微结构的改变.结合SHG无损光学成像技术与HPF的超微结构保留能力,可为相关科学研究提供技术支撑.展开更多
Three-dimensional reconstruction of tissue architecture is crucial for biomedical research.Tissue optical clearing technology overcomes light scattering limitations in biological tissues,providing an essential tool fo...Three-dimensional reconstruction of tissue architecture is crucial for biomedical research.Tissue optical clearing technology overcomes light scattering limitations in biological tissues,providing an essential tool for high-resolution three-dimensional imaging.Given the high degree of similarity between large model animals(e.g.,pigs,non-human primates)and humans in terms of anatomical structure,physiologic function,and disease mechanisms,the application of this technology in these models holds significant value for biomedical research.While well-established tissue clearing protocols exist for tissue sections,whole organs,and even entire bodies in rodents,scaling up to large animal specimens presents substantial challenges due to dimensional effects and compositional variations.This review systematically examines the methodological translation from rodent to large animals,particularly on species-specific differences in brain architecture and parenchymal organ composition that critically impact clearing efficiency.We comprehensively summarize recent applications in large animals,focusing on representative areas including neural circuit mapping,sensory organ imaging,and other related research domains,while proposing optimization strategies to overcome cross-species compatibility barriers.We hope this review will serve as a valuable reference for advancing tissue optical clearing applications in large-animal biomedical research.展开更多
基金the National Nature Science Foundation(Grant Nos.30770552 and 30911120074)the National High-Tech Research and Development Program(No.2008AA02Z107)of China.
文摘In order to enhance the optical clearing effect of topically applied optical clearing agents(OCAs),we evaluated the effect of propylene glycol(PG)as a chemical penetration enhancer(PE)on optical clearing of skin in vitro by observation and measurement of optical-transmittance and diffuse-reflectance spectra.Three OCAs,i.e.,glycerol,D-sorbitol and PEG400,and two other penetration enhancers,Azone and Thiazone,were used in this study.The results indicated that the decrease of reduced scattering coefficient caused by OCA/PG was larger than that by pure OCA,and the change by OCA/water was the least after the same treatment time.There were significant differences for the reduced scattering coefficient at 630 nm after 120 min application of agents between OCA and OCA/PG.The efficacy of optical clearing caused by OCA/PG depended on the OCA itself.When PEG400 was mixed with three different PEs,we found the optical clearing were different.The penetration enhancing ability of PG was much better compared to Azone,and suboptimal to Thiazone.Also,this study provides evidence for the use of PG as a PE in order to improve skin optical clearing.
基金Ths work was supported by the VEKOP-2.3.2-16-2016-00011 Grantwhich is co-financed by the European Union and European Social FundValery V.Tuchin was supported by RFBR Grant 18-52-16025 and the Grant of the Government of the Russian Federation(Registration No.2020-220-08-2389).
文摘Confocal Raman microspectroscopy(CRM)with 633-and 785-nm excitation wavelengths combined with optical clearing(OC)technique was used for ex-vivo study of porcine skin in the Raman fingerprint region.The optical clearing has been performed on the skin samples by applying a mixture of glycerol and distilled water and a mixture of glycerol,distilled water and chemical penetration enhancer dimethyl sulfoxide(DMSO)during 30 min and 60 min of treat-ment.It was shown that the combined use of the optical clearing technique and CRM at 633 nm allowed one to preserve the high probing depth,signal-to-noise ratio and spectral resolution simultaneously.Comparing the effect of different optical clearing agents on porcine skin showed that an optical clearing agent containing chemical penetration enhancer provides higher optical clearing efficiency.Also,an increase in treatment time allows to improve the optical clearing efficiency of both optical clearing agents.As a result of optical clearing,the detection of the amide-Ⅲ spectral region indicating well-distinguishable structural differences between the type-Ⅰ and type-Ⅳ collagens has been improved.
基金supported by research grants from the National Natural Science Foundation of China(Nos.30470426,30870675).
文摘Our previous studies demonstrated the ultrasound-induced skin optical clearing enhancement with topical application of optical clearing agents on in vitro porcine skin and in vivo human skin.The objective of this study was to investigate the possible mechanisms of the enhanced skin optical clearing by ultrasound medications.Optical clearing effects of ex vivo guinea pig abdomen skin topically applied with 60% glycerol or the combination of 60% glycerol and ultrasound were studied by optical coherence tomography(OCT).Microstructure of skin surface was examined by scanning electron microscopy(SEM).Ultrasound with a frequency of 1MHz and a power of 0.75W over a 3-cm probe was simultaneously applied with glycerol solution for 15min.The combination of 60% glycerol and ultrasound results in a 19% increase in OCT 1/e light penetration depth after 30min,which is much better than 60% glycerol alone.SEM images demonstrated that changes in skin microstructure due to the tight order of the lipid bilayers in the stratum corneum disrupted and the separation of keratinocytes by the application of ultrasound contribute to the ultrasound-enhanced intact skin optical clearing effects.
基金supported by the National Natural Science Foundation of China(Grant No.51727811).
文摘The effect of optical cleaning method combined with laser speckle imaging(LSI)was discussed to improve the detection depth of LSI due to high scattering characteristics of skin,which limit its clinical application.A double-layer skin tissue model embedded with a single blood vessel was established,and the Monte Carlo method was used to simulate photon propagation under the action of light-permeating agent.808 nm semiconductor and 632.8 nm He–Ne lasers were selected to study the e®ect of optical clearing agents(OCAs)on photon deposition in tissues.Results show that the photon energy deposition density in the epidermis increases with the amount of tissue°uid replaced by OCA.Compared with glucose solution,polyethylene glycol 400(PEG 400)and glycerol can considerably increase the average penetration depth of photons in the skin tissue,thereby raising the sampling depth of the LSI.After the action of glycerol,PEG 400,and glucose,the average photon penetration depth is increased by 51.78%,51.06%,and 21.51%for 808nm,68.93%,67.94%,and 26.67%for 632.8 nm lasers,respectively.In vivo experiment by dorsal skin chamber proves that glycerol can cause a substantial decrease in blood°ow rate,whereas PEG 400 can signicantly improve the capability of light penetration without a®ecting blood velocity,which exhibits considerable potential in the monitoring of blood°ow in skin tissues.
基金funded in part by NIH BRP grant#1R01HL098912-01by a fellowship for William CVogt provided by the Institute for Critical Technology and Applied Science(ICTAS)Doctoral Scholars Program at Virginia Tech.
文摘Tissue Optical Clearing Devices(TOCDs)have been shown to increase light transmission through mechanically compressed regions of naturally turbid biological tissues.We hypothesize that zones of high compressive strain induced by TOCD pins produce localized water displacement and reversible changes in tissue optical properties.In this paper,we demonstrate a novel combined mechanical finite element model and optical Monte Carlo model which simulates TOCD pin compression of an ex vivo porcine skin sample and modified spatial photon fluence distributions within the tissue.Results of this simulation qualitatively suggest that light transmission through the skin can be significantly affected by changes in compressed tissue geometry as well as concurrent changes in tissue optical properties.The development of a comprehensive multi-domain model of TOCD application to tissues such as skin could ultimately be used as a framework for optimizing future design of TOCDs.
基金supported by the National Natural Science Foundation(Grant Nos.30770552,60828009 and 30911120074)of China.
文摘Laser Speckle Contrast Imaging(LSCI)plays an important role in studying blood flow,but suffers from limited penetration depth of light in turbid tissue.The strong scattering of tissue obviously reduces the image contrast which decreases the sensitivity to flow velocity.Some image processing or optical clearing methods have been proposed to lessen the deficiency,but quantitative assessment of improvement is seldom given.In this study,LSCI was applied to monitor the blood flow through a capillary embedded within various tissue phantoms at depths of 0.25,0.45,0.65,0.85 and 1.05 mm,and the flow velocity in capillary was controllable from 0 to 4mm/s.Here,glycerol,a common optical clearing agent,was mixed with Intralipid at different volume ratio to make the reduced scattering coefficient of tissue phantom decrease from 13.00 to 0.50 cm−1.The quantitative analysis demonstrates that the optical clearing method can obviously enhance the image contrast,imaging depth,and sensitivity to blood flow velocity.Comparing the Laser Speckle Contrast Analysis methods and the optical clearing method,we find that for typical turbid tissue,the sensitivity to velocity estimated by the Laser Speckle Temporal Contrast Analysis(LSTCA)is twice of that by the Laser Speckle Spatial Contrast Analysis(LSSCA);while the sensitivity to velocity estimated by using the two analysis methods has a 10-fold increase,respectively,if addition of glycerol makes the reduced scattering coefficient of tissue phantom decrease by 30%.Combining the LSTCA and the optical clearing method,the sensitivity to flow velocity will be further enhanced.
基金supported by the National Natural Science Foundation of China(NSFC,Nos.30600126,30700168 and 30900275).
文摘Tissue optical clearing by use of optical clearing agents(OCAs)has been proven to have potential to reduce the highly scattering effect of biological tissues in optical techniques.However,the difference in tissue samples could lead to unreliable results,making it difficult to quantitatively control the dose of OCAs during the course of tissue optical clearing.In this work,in order to study the effects of optical clearing,we customized tissue-like phantoms with optical properties of some biological tissue.Diffuse reflectance and total transmittance of tissue-like phantoms with different OCAs(DMSO or glycerol)and porcine skin tissues were measured.Then optical property parameters were calculated by inverse adding-doubling(IAD)algorithm.Results showed that OCAs could lead to a reduction in scattering of tissue-like phantoms as it did to porcine skin tissue in vitro.Furthermore,a series of relational expressions could be fit to quantitatively describe the relationship between the doses of OCAs and the reduction of scattering effects.Therefore,proper tissue-like phantom could facilitate optical clearing to be used in quantitative control of tissue optical properties,and further promote the application potential of optical clearing to light-based noninvasive diagnostic and therapeutic techniques.
基金support from the National Key Research and Development Program of China(Grant No.2017YFA0700501),and the Innovation Fund of WNLO.
文摘Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness.While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D,they often have limitations,such as being too harsh for fragile 3D cell cultures,requiring complex handling protocols,or inducing tissue deformation with shrinkage or expansion.To address this issue,we proposed a modified optical clearing method for 3D cell cultures,called MACS-W,which is simple,highly efficient,and morphology-preserving.In our evaluation of MACS-W,we found that it exhibits excellent clearing capability in just 10 min,with minimal deformation,and helps drug evaluation on tumor spheroids.In summary,MACS-W is a fast,minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures.
文摘For the first time,the changes in autofluorescence spectra of ex vivo rat skin have been experimentally investigated using the combination of fluorescence spectroscopy and optical immersion clearing.The glucose,glycerol and propylene glycol solutions were used as clearing agents.The optical clearing was performed from the dermal side of skin imitating the in vivo injection of clearing agent under the dermal layers.In this contribution,the common properties of autofluorescence variation during optical immersion clearing were determined.The tendency of autofluorescence signal to decrease with reduction of scattering in tissue was noticed and discussed in detail.However,the differences in the shape of spectral curves under application of different clearing agents showed that optical clearing affects the autofluorescence properties of tissue differently depending on the type of clearing liquid.The results obtained are useful for the understanding of tissue optical clearing mechanisms and for improving techniques such as fluorescence spectroscopy.
基金supported by the National Nature Science Foundation of China (Grant Nos.81171376,91232710,812111313)the Science Fund for Creative Research Group (Grant No.61121004)the Research Fund for the Doctoral Program of Higher Education of China (Grant No.20110142110073).
文摘Lasers have been widely used for tattoo removal,but the limited light penetration depth calused by high skin scattering property restricts the therapeutic outcome of deep tattoo.Skin optical clearing method,by introducing optical clearing agent(OCA)into skin,has shown some im-provement in the effect of laser tattoo removal.In this study,the enhanced laser tattoo removal has been quantitatively assessed.OCA was applied to the skin of tattoo animal model and Q switched Nd:YAG laser(1064 nm)irradiation was used to remove the tattoo.The skin evaluation instrument(Mexameter probe,MPA580)was applied to measure the content of tattoo pigment before and after laser treatment,and then the clearance rate of pigment was calculated.Further,Monte Carlo(MC)method was utilized to simulate the efecet of skin optical clearing on light transmission in tattoo skin model.By comparing the pigment change of tattoo areas respectively treated with OCA plus laser and single laser,it was found that pigment clearance of the former tattoo area was increased by 1.5-fold.Further,the MC simulation verifed that the reduced light scattering in skin could increase the effective dose of lumninous fux reaching to the deep tattoo regions.It can be concluded from both experiment and theoretical simulations that skin optical clearing technique could improve the outcome of laser tattoo re moval,which should be beneficial for clinical laser tattoo removal and other laser pigment elimination.
基金supported by the National Natural Science Foundation of China(Grant Numbers 62205025 and 61835015)Beijing Natural Science Foundation(7222309)+2 种基金the Open Project Program of Wuhan National Laboratory for Optoelectronics(2020WNLOKF025)CAMS Innovation Fund for Medical Sciences(CIFMS)(2019-I2M-5-061)Beijing Institute of Technology Research Fund Program for Young Scholars(XSQD-202123001).
文摘Vascular-targeted photodynamic therapy(V-PDT)is an effective treatment for port wine stains(PWS).However,repeated treatment is usually needed to achieve optimal treatment outcomes,possibly due to the limited treatment light penetration depth in the PWS lesion.The optical clearing technique can increase light penetration in depth by reducing light scattering.This study aimed to investigate the V-PDT in combination with an optical clearing agent(OCA)for the therapeutic enhancement of V-PDT in the rodent skinfold window chamber model.Vascular responses were closely monitored with laser speckle contrast imaging(LSCI),optical coherence tomography angiography,and stereo microscope before,during,and after the treatment.We further quantitatively demonstrated the effects of V-PDT in combination with OCA on the blood flow and blood vessel size of skin microvasculature.The combination of OCA and V-PDT resulted in significant vascular damage,including vasoconstriction and the reduction of blood flow.Our results indicate the promising potential of OCA for enhancing V-PDT for treating vascular-related diseases,including PWS.
基金supported by grants from the National Key Research and Development Program of China(No.2016YFC1101604)the Fundamental Research Funds for the Central Universities+2 种基金Clinical Medicine Plus X-Young Scholars Project of Peking University China(No.PKU2020LCXQ020)Guangdong Basic and Applied Basic Research Foundation(No.2019A1515110983,No.2021A1515012586)Bethune Charitable Foundation and CSPC Osteoporosis Research Foundation Project(No.G-X-2020-1107-21).
文摘Objective:Wallerian degeneration is a pathological process closely related to peripheral nerve regeneration following injury,and includes the disintegration and phagocytosis of peripheral nervous system cells.Traditionally,morphological changes are observed by performing immunofluorescence staining after sectioning,which results in the loss of some histological information.The purpose of this study was to explore a new,nondestmetive,and systematic method for observing axonal histological changes during Wallerian degeneration.Methods:Thirty male Thy1-YFP-16 mice(SPF grade,6 weeks old,20±5 g)were randomly selected and divided into clear,unobstructed brain imaging cocktails and computational analysis(CUBIC)optical clearing(n=15)and traditional method groups(n=15).Five mice in each group were sacrificed at 1st,3rd,and 5th day following a crush operation.The histological axon changes were observed by CUBIC light optical clearing treatment,direct tissue section imaging,and HE staining.Results:The results revealed that,compared with traditional imaging methods,there was no physical damage to the samples,which allowed for three-dimensional and deep-seated tissue imaging through CUBIC.Local image information could be nicely obtained by direct fluorescence imaging and HE staining,but it was difficult to obtain image information of the entire sample.At the same time,the image information obtained by fluorescence imaging and HE staining was partially lost.Conclusion:The combining of CUBIC and Thy1-YFP transgenic mice allowed for a clear and comprehensive observation of histological changes of axons in Wallerian degeneration.
基金funded by the National Nature Science Foundation of China(grant nos.62375096,61860206009,32361133552,and 82361138569)the Russian Science Foundation(grant no.24-44-00082)。
文摘Organisms are composed of a vast number of highly specialized cells with characteristic physiological functions determined by their structure.Therefore,understanding physiological and pathological mechanisms requires obtaining 3-dimensional(3D)structural information at the cellular level across tissues,organs,and even the whole body.However,the scattering and absorption of tissue limits the penetration depth of light and thus extremely reduces the imaging performance in deep tissue.It is well known that 3D imaging outperforms 2D imaging in various scenarios for 3D reconstruction of tissue structures.This includes tracing convoluted 3D structures like vasculature,neurons,and prostate glands,as well as elucidating intricate distributions of cells or lesions.
基金This work was carried out under the support from the Russian Foundation for Basic Research(Grant Nos.18-52-16025 and 19-32-90224)The Fund for Promoting Innovation grant UMNIK-19/HealthNet.NTI-2019 No.15929GU/2020 of 07.23.2020(code 0059878,application(U-65096)).
文摘Complex study of glycerol effects on the skin tissue was performed.The change in optical,weight and geometrical parameters of the rat skin under the action of the glycerol solutions was studied ex vivo.Possible mechanisms of the skin optical clearing under the action of glycerol solutions of different concentrations were discussed.The results can be helpful for refinement of models developed to evaluate the effective diffusion coefficients of glycerol in tissues.
基金supported in part by grants:RF Governmental contracts 11.519.11.2035,14.B37.21.0728,and 14.B37.11.0563FiDiPro,TEKES Program(40111/11),Finland+1 种基金SCOPES EC,Uzb/Switz/RF,Swiss NSF,IZ74ZO 137423/1RF President's grant"Scientific Schools",1177.2012.2.
文摘With the objective to study the variation of optical properties of rat muscle during optical clearing,we have performed a set of optical measurements from that kind of tissue.The.measurements performed were total tr ansmittance,ollimated transmit tance,specular reflec-tance and total reflectance.This set of measurements is suficient to determine diffuse reflectance and absorbance of the sample,also necessary to est imate the optical properties.All the per formed measurements and calculated quantities will be used later in inverse Monte Carlo(IMC)simu-lations to determine the evolution of the optical properties of muscle during treatments with ethylene glycol and glucose.The results obt ained with the measurements already provide some information about the optical c learing treatments applied to the muscle and translate the mechanisms of turning the tissue more transparent and sequence of regimes of optical clearing.
文摘以甘油光学清除的小鼠皮肤为研究对象,通过二次谐波(second-harmonic generation,SHG)成像和透射电子显微镜(transmission electron microscope,TEM)研究光学清除对组织超微结构的影响.SHG成像结果表明,甘油处理能够显著增大信号强度并提升成像深度,证实其可作为一种有效的光学清除剂.通过TEM对比观察常规化学固定与高压冷冻(high-pressure freezing,HPF)固定样品的超微结构,发现HPF能更好地保持组织自然状态.进一步分析显示,甘油处理虽使胶原纤维直径减小,但其排列周期未发生明显变化,说明光学清除过程未对胶原纤维结构造成损伤.上述结果表明,甘油具有高效、安全的光学清除效果,且在光学清除过程中不会引起组织超微结构的改变.结合SHG无损光学成像技术与HPF的超微结构保留能力,可为相关科学研究提供技术支撑.
基金supported by the National Natural Science Foundation of China(Grant Nos.62375096,82361138569,82372012,82402341)the Open Competition Project of Wuhan East Lake High-tech Development Zone(Grant No.2023KJB224)the Innovation Project of Optics Valley Laboratory(Grant No.OVL2025BB008).
文摘Three-dimensional reconstruction of tissue architecture is crucial for biomedical research.Tissue optical clearing technology overcomes light scattering limitations in biological tissues,providing an essential tool for high-resolution three-dimensional imaging.Given the high degree of similarity between large model animals(e.g.,pigs,non-human primates)and humans in terms of anatomical structure,physiologic function,and disease mechanisms,the application of this technology in these models holds significant value for biomedical research.While well-established tissue clearing protocols exist for tissue sections,whole organs,and even entire bodies in rodents,scaling up to large animal specimens presents substantial challenges due to dimensional effects and compositional variations.This review systematically examines the methodological translation from rodent to large animals,particularly on species-specific differences in brain architecture and parenchymal organ composition that critically impact clearing efficiency.We comprehensively summarize recent applications in large animals,focusing on representative areas including neural circuit mapping,sensory organ imaging,and other related research domains,while proposing optimization strategies to overcome cross-species compatibility barriers.We hope this review will serve as a valuable reference for advancing tissue optical clearing applications in large-animal biomedical research.
