Absin多重荧光免疫组化服务

时间：2022-10-28 点击次数：6415

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【技术原理】

多重荧光免疫组化（mIHC），主要的技术原理来自TSA技术，TSA即酪酰胺信号放大（Tyramide signal amplification）是一类利用辣根过氧化酶（HRP）对靶蛋白或核酸进行高密度原位标记的酶学检测方法。该技术可与高性能染料Alexa Fluor®、传统荧光染料和比色检测系统相兼容。

TSA主要原理是利用酪胺 Tyramide 的过氧化物酶反应（酪胺盐在HRP催化H₂O₂下形成共价键结合位点），产生大量的酶促产物，该产物能与周围的蛋白残基（包括色氨酸、组氨酸和酪氨酸残基）结合，这样在抗原-抗体结合部位就有大量的生物素沉积，与随后加入的 Streptavidin—HRP/荧光基团结合，经几次这样的循环放大，可以结合大量的酶分子or荧光基团，结果使其检测信号得到几何级放大。

TSA多重荧光免疫组化染色示意图

简单来说，用这种方法做多重荧光染色是利用二抗上带有的HRP（而不是直接偶联荧光素），来催化后续添加入体系的非活性荧光素。该荧光素在HRP和过氧化氢的作用下被活化，跟临近蛋白的酪氨酸残基共价偶联，使得蛋白样品与荧光素稳定结合。然后进行微波处理，前一轮非共价结合的抗体被洗掉，共价结合的荧光素还留存在样品上。再换个一抗来第二轮孵育，周而复始。等到所有抗体孵育结束，荧光素都结合好后，最后去检测结果。

【mIHC技术优势】

1. 由于染色是多轮次进行，每次体系中都只有单一抗体孵育，因此无需担心抗体交叉反应，以及一抗二抗种属匹配问题，大大减少了实验设计时不同种属抗体选择匹配的限制。

2. TSA技术可以用于检测用传统方法无法检出的低丰度靶标。基于酪酰胺的信号放大技术能够提供极强的敏感度、检测极微量的目的抗原。极大的降低抗体的用量，节约抗体，实测确实可以显著提升抗体的稀释比例。

3. 多重荧光免疫组化法，可同时进行九个指标十色成像，这种情况下发射光谱会重叠，我们可以借助多光谱成像系统来解决这个问题。这是传统间接荧光法染色和显色法多重免疫组化所做不到的。这种方法能真实反映关键蛋白的表达情况和相互之间的关系，从而极大地节约珍贵的样品。

【Absin平台优势】

1. Absin与TISSUEGNOSTICS两大公司强强联手，共建联合实验室。Absin拥有10000+组织切片及1000+指标的多重荧光免疫组化染色经验，在评估肿瘤微环境中各细胞组分的表达丰度，相互作用以及空间表型信息等领域深耕多年；TG TissueFAXs Spectra Systems全景多光谱组织扫描与定量分析系统，能够实现大尺寸高倍率全自动连续光谱成像及可原位追溯的大数据Al深度空间量化分析功能；

2. 兼有3DHISTECH PANNORAMIC MIDI II扫描仪、HALO数字病理金标准分析平台，满足您的多样化需求；

3. 资深多色团队共同服务客户，为您提供常见免疫微环境panel推荐、抗体推荐、项目探讨等售前售后咨询服务。

【爱必信多色服务】

我们可以提供的服务：

最多九指标十色（含DAPI）的多重荧光免疫组化服务，样本种属涵盖人、小鼠、大鼠、猪等；组织类型以肿瘤样本居多，涵盖肺、胃、肝、肾、肠等各大器官，甲状腺、胰腺、唇腺、泪腺等各大腺体，另还有眼球、神经、大脑等特殊样本；切片类型涵盖石蜡切片、组织芯片（TMA）、冰冻切片、大组织切片、厚切片等，并提供全片扫描和定量分析。

个性化服务（先做预实验，实验有保障，结果更完美！）

标准化检测（高性价比，已优化过条件，只需提供样本，直接出实验结果！强烈推荐！）

Absin Flash Panel
人	小鼠
T细胞：CD3、CD4、CD8、FOXP3、CD45、CD103...	T细胞：CD3、CD4、CD8、FOXP3、CD45....
巨噬细胞：CD68、CD163、CD206、CD86...	巨噬细胞：F4/80、CD86、CD11b、CD163、M-CSF...
免疫检查点：PD-1、PD-L1、HLA-DR、TIM-3、GZMB、CD24、LAG-3...	免疫检查点：GZMB、PD-1、PD-L1
肿瘤相关：Ki-67、PANCK、VEGF...	肿瘤相关：SHP-2...
NK细胞：CD56...	NK细胞：NKR-P1C（NK1.1）...
中性粒细胞：CD11c...	中性粒细胞：LY6G、CD11c...
成纤维细胞：α-SMA...	信号通路相关：Arrestin-C、iNOS...
B细胞：CD20、CD19...	白介素：IL-4、IL-10、IL-13...

Do It Yourself！Absin精选多色试剂盒（有仪器，自己做，操作简便，染料二抗一应俱全！）

Multicolor-Kit	染料组成	其他成分	备注
四色多重荧光免疫组化试剂盒	TSA520/TSA570/TSA650	信号放大反应液鼠兔通用/抗兔二抗封片剂 DAPI	常规拥有DAPI，FITC，CY3，CY5等通道的荧光/共聚焦显微镜即可成像
五色多重荧光免疫组化试剂盒	TSA520/TSA570/TSA620/TSA700		常规拥有DAPI，FITC，CY3，CY5等通道的荧光/共聚焦显微镜即可成像
六色多重荧光免疫组化试剂盒	TSA520/TSA570/TSA620/TSA650/TSA700		需要仪器加装有窄带滤光块或仪器具有多光谱光谱拆分功能可成像
七色多重荧光免疫组化试剂盒	TSA520/TSA540/TSA570/TSA620/TSA650/TSA700
七色多重荧光免疫组化试剂盒（plus）	TSA520/TSA700/TSA570/TSA620/TSA480/TSA770

我们可以提供的服务结果：

1. 全片200X（相当于物镜20X，目镜10X）原图，提供看图软件可在客户端打开。数据上传百度网盘。

2. 客户指定区域的单细胞分析结果：
单阳性、双阳性或三阳性细胞占有核细胞百分比；
组织面积和特定病理形态面积计算；
TMA提供每个芯点的阳性细胞百分比及面积计算，并提供热图。

【图片风采展】

1. 肺鳞癌切片/类器官肿瘤标志物检测

2. 甲状腺七色免疫微环境检测 CD4、FOXP3、CD20、CD56、CD3、panCK

The immune-suppressive landscape in lepromatous leprosy revealed by single-cell RNA sequencing. IF38.079

The thermogenic activity of adjacent adipocytes fuels the progression of ccRCC and compromises anti-tumor therapeutic efficacy. IF27.281

Chemical-induced chromatin remodeling reprograms mouse ESCs to totipotent-like stem cells. IF24.633

Runt-related transcription factor-1 ameliorates bile acid-induced hepatic inflammation in cholestasis through JAK/STAT3 signaling. IF17.298

CAFs-derived SCUBE1 promotes malignancy and stemness through the Shh/Gli1 pathway in hepatocellular carcinoma. IF:8.440

mTOR–neuropeptide Y signaling sensitizes nociceptors to drive neuropathic pain. IF:9.484

Single-cell RNA sequencing reveals cellular and molecular reprograming landscape of gliomas and lung cancer brain metastases. IF=8.554

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