上海信帆生物带您了解：组织自发荧光淬灭剂 

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组织自发荧光淬灭剂 

描述：许多组织会产生可透过各种波长滤光片的组织内源性自发荧光，显著干扰抗体标记荧光观察甚至导致荧光组化染色失败。自发荧光淬灭试剂中的离子可用碰撞方式捕获自发荧光光源分子发出的电子，阻止该电子从激发态回归基态并阻止能量释放，从而淬灭自发荧光。采用优化的孵育时间，可以大限度地消除自发荧光而不明显影响抗体标记的荧光。

适用：各种组织、细胞免疫荧光染色的自发荧光消除。特别适用于神经组织自发荧光淬灭。

储存：4 ºC避光。

用法：

以下步骤在免疫荧光组化染色完毕之后(而非在荧光染色完毕之前)执行。对特定的组织和细胞类型，必需优化孵育时间以便大限度淬灭自发荧光而不明显影响抗体标记的荧光(步骤2)。仔细阅读后面说明。

1. 吸去PBS或相应洗涤缓冲液，用蒸馏水短暂冲洗组织切片或细胞培养板中的细胞。

2. 加入适量但充足的自发荧光淬灭剂覆盖组织切片或瓶皿中的细胞。室温10-90min。

3. 吸去自发荧光淬灭剂，用蒸馏水短暂冲洗。

4. 吸去蒸馏水，用PBS覆盖组织切片或位于细胞培养板中的细胞。

5. 封片。建议使用抗荧光衰减封片剂。该封片剂可防止抗体标记荧光衰退。

6. 荧光显微镜观察。

说明：

1.不同物种不同类型的组织的自发荧光具有不同的特征，使用组织自发荧光淬灭剂的效果可能会有差别。另外，任何针对自发荧光的淬灭，将会在一定程度上降低抗体荧光强度。所幸的是该试剂对自发荧光的淬灭程度远远超出抗体荧光强度的降低，因而能在二者之间获得较好的平衡。由于不很清楚的原因，本试剂消除脑脊髓神经组织的自发荧光具有更好的效果。

2. 为获得佳效果，必需优化孵育时间以便大限度淬灭对某一特定组织的自发荧光而不明显影响抗体标记的荧光(步骤2)。重要的标本应在确定佳孵育时间之后使用本试剂。进行优化时，可取数张组织切片或位于培养皿中的细胞，在免疫荧光组化染色完毕之后加入组织自发荧光淬灭剂，孵育5、10、30、60、90分钟等不同时间，冲洗后观察荧光。如果组织自发荧光仍然很强，可延长孵育时间；如果孵育时间小于10分钟而荧光消退十分明显，可将孵育时间减少为1-5分钟，或者可取出少量的组织自发荧光淬灭剂加等份的双蒸水稀释，然后孵育10-90分钟优化。

3. 组织自发荧光淬灭剂必须在完成免疫荧光组化染色后使用，否则将严重降低抗体荧光。

AutoFluo Quencher

Description:

Many tissues can produce endogenous spontaneous fluorescence which can be observed through various wavelength filters, which can obviously interfere with antibody labeling fluorescence and even lead to the failure of fluorescence histochemical staining. The ions in the spontaneous fluorescence quenching reagent can capture the electrons emitted by the spontaneous fluorescent light source molecules in a collision mode, which prevents the electron from returning to the ground state and prevents the energy release from the excited state, thereby quenching the spontaneous fluorescence. By optimizing the incubation time, the fluorescence of the antibody could be eliminated and the fluorescence was not obviously affected by the spontaneous fluorescence. 

Application: spontaneous fluorescence elimination of various tissues and cells by immunofluorescence staining. In particular, the application of neural tissue spontaneous fluorescence quenching.

Storage at :  4 degrees C & keep away from light.

Usage:

The following steps are performed after the IHC staining (and not before the fluorescent staining). For specific tissues and cell types, it is necessary to optimize the incubation time in order to maximize the quenching of the fluorescence (step 2), which is not significantly affected by the antibody labeling. Read the instructions carefully.

1 suction to PBS or the corresponding washing buffer, using distilled water briefly washed tissue sections or cell culture plate in the cell.

2 adding adequate amount of spontaneous fluorescent quenching agent to cover the cells in tissue sections or bottles. Room temperature 10-90min.

3 to absorb the spontaneous fluorescence quenching agent, with distilled water briefly rinse.

4 suck the distilled water, cover the tissue sections with PBS or cells in the cell culture plate.

5 mounting. Anti sealing agents using the proposed fluorescence decay. The sealing agents can prevent the antibody labeled with fluorescent decay.

6 fluorescence microscope observation.

Note：

1. different types of tissues of different types of spontaneous fluorescence with different characteristics, the use of spontaneous fluorescence quenching agent effect may be different. In addition, any quenching of spontaneous fluorescence will reduce the fluorescence intensity of antibody to a certain extent. Fortunately, the quenching degree of the reagent is far more than the decrease of the fluorescence intensity of the antibody, so that a good balance between the two can be obtained. Because of the unclear reasons, this reagent eliminates the spontaneous fluorescence of the spinal cord nerve tissue and has a better effect.

2. in order to obtain the best results, it is necessary to optimize the incubation time in order to maximize the quenching of the spontaneous fluorescence of a particular tissue and not to affect the fluorescence (step 2). Important specimens should be used to determine the optimal incubation time. Optimize, the number of desirable tissue section or in the cells in a Petri dish, after immunohistochemistry after adding tissue fluorescence quenching agent, were incubated for 5, 10, 30, 60, 90 minutes of different time, observe the fluorescence after washing. If the tissue autofluorescence is still strong, can prolong the incubation time; if the incubation time is less than 10 minutes and fluorescence fade is very obvious, can reduce the incubation time for 1-5 minutes, or you can remove a small amount of tissue autofluorescence quenching agent with equal distilled water dilution, and then incubated for 10-90 minutes optimization.

3. tissue spontaneous fluorescent quenching agent must be used after the completion of immunohistochemical staining, otherwise it will seriously reduce the antibody fluorescence.

上海信帆生物带您了解：组织自发荧光淬灭剂