A multispectral LED array for the reduction of background autofluorescence in brain tissue.
A multispectral LED array for the reduction of background autofluorescence in brain tissue.
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DOI：
10.1016/j.jneumeth.2013.08.018
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发表时间：
2013-10-30
影响因子：
3
通讯作者：
Han, Martin
中科院分区：
文献类型：
--
作者：
Duong, Haison;Han, Martin
关键词：
固定剂诱导的和细胞衍生的人工自发荧光（AAF）的存在对涉及免疫荧光的组织学分析提出了挑战。我们已经建立了一个简单而高效的方法来减少AAF，这是普遍存在于固定的哺乳动物大脑和其他组织。使用市售的发光二极管（LED）阵列、冷却单元和支撑部件构造紧凑的AAF猝灭光照射装置。LED面板由多光谱高强度LED阵列组成，其用作光照射过程的照明源。分别对5 μm和40 μ m厚的兔脑和猫脑标本进行不同时间的光照射。处理24 h后，脱蜡和石蜡化兔脑标本以及自由漂浮猫脑标本的AAF均降低至接近组织背景水平。随后使用针对GFAP、NeuN、Iba-1和MAP-2的一抗以及相应的Qdot®和Alexandor ®荧光缀合物的免疫荧光染色证实，LED光照射处理不损害免疫荧光标记的效率。该装置的使用不是劳动密集型的，并且仅需要最少的组织处理和实验设置时间，具有非常低的维护和操作成本。最后，多个标本，在两个幻灯片和孔板格式，可以同时进行光照射，因此，允许可扩展的批量处理。
The presence of fixative-induced and cellular-derived artifactual autofluorescences (AAFs) presents a challenge in histological analysis involving immunofluorescence. We have established a simple and highly effective method for the reduction of AAFs that are ubiquitous in fixed mammalian brain and other tissues. A compact AAF-quenching photo-irradiation device was constructed using a commercially available light emitting diode (LED) array, cooling unit, and supporting components. The LED panel is comprised of an array of multispectral high intensity LEDs which serve as the illumination source for the photo-irradiation process. Rabbit and cat brain specimens of 5 μm- and 40 μm-thicknesses, respectively, were photo-irradiated for various durations. The AAFs were reduced to near tissue background levels after 24 h of treatment for both deparaffinized and paraffinized rabbit brain specimens, and for the free-floating cat brain specimens. Subsequent immunofluorescence staining using primary antibodies against GFAP, NeuN, Iba-1, and MAP-2, and the corresponding Qdot® and Alexafluor® fluoroconjugates confirmed that the LED photo-irradiation treatment did not compromise the efficiency of the immunofluorescence labeling. The use of the device is not labor intensive, and only requires minimal tissue processing and experimental set-up time, with very low maintenance and operating costs. Finally, multiple specimens, in both slide and well-plate format, can be simultaneously photo-irradiated, thus, allowing for scalable batch processing.
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影响因子：
4.1
作者：
OCONNELL, MJ;BAUM, H;PETERS, TJ
通讯作者：
PETERS, TJ
影响因子：
3.2
作者：
Schnell, SA;Staines, WA;Wessendorf, MW
通讯作者：
Wessendorf, MW
影响因子：
3
作者：
Potter, Kelsey A.;Simon, Joel S.;Capadona, Jeffrey R.
通讯作者：
Capadona, Jeffrey R.
影响因子：
3.2
作者：
Neumann, M;Gabel, D
通讯作者：
Gabel, D
DOI：
10.1109/tbme.2011.2172440
发表时间：
2012-02
期刊：
IEEE transactions on bio-medical engineering
影响因子：
--
作者：
Han M;Manoonkitiwongsa PS;Wang CX;McCreery DB
通讯作者：
McCreery DB