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Method Article
The assembly and use of a multimodal microendoscope is described which can co-register superficial tissue image data with tissue physiological parameters including hemoglobin concentration, melanin concentration, and oxygen saturation. This technique can be useful for evaluating tissue structure and perfusion, and can be optimized for individual needs of the investigator.
最近纤维束显微内窥镜技术使用两种成像技术或分光技术的组合在体内组织的非侵入性的分析。结合成像和光谱学技术成一个单一的光学探针可提供组织健康的更完整的分析。在这篇文章中,两种不同的方式组合,高分辨率荧光显微内窥镜成像和漫反射光谱,成一个单一的光学探头。高分辨率荧光显微内窥镜成像是用于可视化根尖组织微结构的技术,虽然大多是定性的技术,已经证明肿瘤和非肿瘤组织之间的有效实时分化。漫反射光谱是一种技术,它可以提取组织的生理参数,包括当地的血红蛋白浓度,黑色素浓度和血氧饱和度。本文介绍的规格řequired构建光纤探头,如何建立仪器仪表,然后演示了在体内对人体皮肤的技术。这项工作表明该组织的微架构,特别是心尖皮肤角质,可以和其相关的生理参数共同注册。这里介绍的仪器和纤维束探针可以被优化为在各种器官系统的使用任一手持或内窥镜兼容设备。另外的临床研究来测试此技术对于不同的上皮疾病状态的可行性。
纤维束显微内窥镜技术通常使用两种成像技术或分光技术的组合分析体内组织中 1-3一种这样的成像技术,高分辨率荧光显微内窥镜,可以形象根尖组织微架构具有亚细胞分辨率的小,微尺度场的图,利用局部造影剂如原黄素,荧光素,或羟基芘磺酸油墨。1,3-11该成像模态已示于定性实时低区分患病的和健康上皮组织有前途的临床表现间观察员变异8偶尔,调查人员将使用高分辨率荧光显微镜的数据中提取定量的功能,如细胞与核的大小或腺区,但这仍然瞄准往可视化的组织形态的主要定性技术。1,3,8- 10在另一方面,光谱技术等作为漫反射光谱,正朝着提供功能组织信息并显示在多个器官定量识别病变癌变看好的临床表现有针对性的。2,12-15
因此,有必要对掺入两种类型的模式,从而可能进一步降低观察者间变异性,维持组织微架构的实时可视化,并提供组织健康的一个更完整的分析的装置。为了实现这个目标,在基于探针的多峰仪器构建结合在一个单一的光纤探针两种模式:高分辨率荧光显微内窥镜和分漫反射光谱11此方法共寄存器定性心尖的高清晰度图像组织形态(结构特性)与来自两个不同的组织深度定量光谱信息(功能性),包括局部血红蛋白浓度([血红蛋白]),黑色素浓度([梅尔])和氧饱和度(SAO 2)。11,12,16这个特定副漫反射光谱....
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在阿肯色大学从人类受试者的研究项目,获得了本次研究的方方面面机构审查委员会的批准(IRB#15-09-149)。所描述的方法是在按照批准的指导方针进行,并从所有参与者获得知情同意书。
1.高分辨率荧光显微内窥镜成像设备的组装
注意:对于高分辨率荧光显微内窥镜形态的组件的概述的步骤可以在图2中被可视化。
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按照此协议,研究者将获得组织部位的在焦高分辨率图像与全视场( 图5)。细胞的轮廓可以看出,如果沾有墨水羟基芘磺酸从一个黄色荧光笔,而单个细胞核,如果能与染料染色,如原黄素可以看出。下列光谱采集,后处理软件使用体内血红蛋白浓度([血红蛋白])和黑素浓度([梅尔])21的先验知识,以适应子漫反射光谱,并确定为[血.......
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多峰高分辨率成像和分漫反射光谱纤维束显微内窥镜此处报告可以被优化并用于各种应用,包括内窥镜或手持使用用于人类或动物研究中使用由调查员。因此,它提供了用于在体内根尖组织微结构可视化与来自两个不同的组织深度的血红蛋白浓度,黑色素浓度和组织氧饱和度的测量值的灵活的方法。本文介绍了用于光纤探针的说明书,所述的协议用于组装高分辨率成像系统和分漫反射?.......
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The authors declare that they have no competing financial interests.
This material is based on work supported by the National Institutes of Health (1R03-CA182052, 1R15-CA202662), the National Science Foundation Graduate Research Fellowship Program (G.G., DGE-1450079), the Arkansas Biosciences Institute, and the University of Arkansas Doctoral Academy Fellowship. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the acknowledged funding agencies.
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Name | Company | Catalog Number | Comments |
30 mm Cage Cube with Dichroic Filter Mount | Thorlabs, Inc. | CM1-DCH | |
470 nm Dichroic Mirror (Beam Splitter) | Chroma Corporation | T470lpxr | |
Cage Assembly Rod, 1.5", 4-Pack | Thorlabs, Inc. | ER1.5-P4 | |
Cage Assembly Rod, 3.0", 4-Pack | Thorlabs, Inc. | ER3-P4 | |
Cage Assembly Rod, 2.0", 4-Pack | Thorlabs, Inc. | ER2-P4 | |
SM1-Threaded 30 mm Cage Plate | Thorlabs, Inc. | CP02 | |
SM1 Series Stress-Free Retaining Ring | Thorlabs, Inc. | SM1PRR | |
SM1 Lens Tube, 1.00" Thread Depth | Thorlabs, Inc. | SM1L10 | |
Right-Angle Kinematic Mirror Mount | Thorlabs, Inc. | KCB1 | |
1" UV Enhanced Aluminum Mirror | Thorlabs, Inc. | PF10-03-F01 | |
Z-Axis Translation Mount | Thorlabs, Inc. | SM1Z | |
10X Olympus Plan Achromatic Objective | Thorlabs, Inc. | RMS10X | |
XY Translating Lens Mount | Thorlabs, Inc. | CXY1 | |
SMA Fiber Adapter Plate with SM1 Thread | Thorlabs, Inc. | SM1SMA | |
SM1 Lens Tube, 0.50" Thread Depth | Thorlabs, Inc. | SM1L05 | |
440/40 Bandpass Filter (Excitation) | Chroma Corporation | ET440/40x | |
525/36 Bandpass Filter (Emission) | Chroma Corporation | ET525/36m | |
Quick Set Epoxy | Loctite | 1395391 | |
455 nm LED Light Housing Kit - 3-Watt | LED Supply | ALK-LH-3W-KIT | |
1" Achromatic Doublet, f = 50 mm | Thorlabs, Inc. | AC254-050-A | |
Flea 3 USB Monochrome Camera | Point Grey, Inc. | FL3-U3-32S2M-CS | |
0.5" Post Holder, L = 1.5" | Thorlabs, Inc. | PH1.5 | |
0.5" Optical Post, L = 4.0" | Thorlabs, Inc. | TR4 | |
Mounting Base, 1" x 2.3" x 3/8" | Thorlabs, Inc. | BA1S | |
Long Lifetime Tungsten-Halogen Light Source (Vis-NIR) | Ocean Optics | HL-2000-LL | |
20X Olympus Plan Objective | Edmund Optics, Inc. | PLN20X | |
Custom-Built Aluminum Motor Arm | N/A | N/A | Custom designed and built |
Custom-Built Aluminum Motor Arm Adaptor | N/A | N/A | Custom designed and built |
Custom-Built Aluminum Motor Housing | N/A | N/A | Custom designed and built |
Stepper Motor - 400 steps/revolution | SparkFun Electronics | ROB-10846 | Multiple suppliers |
Custom-Built Aluminum Optical Fiber Switch | N/A | N/A | Custom designed and built |
Custom-Built Aluminum Optical Fiber Switch Face-Plate | N/A | N/A | Custom designed and built |
Arduino Uno - R3 | SparkFun Electronics | DEV-11021 | Multiple suppliers |
Electronic Breadboard - Self-Adhesive | SparkFun Electronics | PRT-12002 | Multiple suppliers |
EasyDriver - Stepper Motor Driver | Sparkfun Electronics | ROB-12779 | |
12 V, 229 mA Power Supply | Phihong | PSM03A | Multiple suppliers |
Enhanced Sensitivity USB Spectrometer (Vis-NIR) | Ocean Optics | USB2000+VIS-NIR-ES | |
550 µm, 0.22 NA, SMA-SMA Fiber Patch Cable | Thorlabs, Inc. | M37L01 | |
Custom-Built Fiber-Optic Probe | Myriad Fiber Imaging | N/A | |
20% Spectralon Diffuse Reflectance Standard | Labsphere, Inc. | SRS-20-010 | |
Standard Yellow Highlighter | Sharpie | 25005 | Multiple suppliers, proflavine or fluorescein can be substituted |
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