这关键区别在直接和间接免疫荧光之间直接免疫荧光使用一个抗体这违反了感兴趣的目标,而间接免疫荧光使用两种抗体来标记感兴趣的靶标。
Immunofluorescence或细胞成像是一种用于标记特定靶抗原的技术荧光团。在这里,荧光团是一种荧光化合物,可以在光激发后重新发射发光。当荧光团与靶抗原结合时,它允许检测样品中的靶分子。为了进一步描述,当抗原与特定抗体结合时,它可以与荧光团结合。因此,在荧光显微镜下观察时,很容易检测样品中靶抗原的存在。
此外,有两种类型的免疫荧光。直接和间接免疫荧光。直接免疫荧光和间接免疫荧光之间的差异主要在于使用的抗体数量和荧光团结合。也就是说,在直接免疫荧光中,荧光团直接与原代抗体结合,而在间接免疫荧光中,荧光团结合了二抗。
CONTENTS
1。概述和关键差异
2。什么是直接免疫荧光
3。什么是间接免疫荧光
4。Similarities Between Direct and Indirect Immunofluorescence
5。Side by Side Comparison – Direct vs Indirect Immunofluorescence in Tabular Form
6.Summary
什么是直接免疫荧光?
免疫荧光使用抗体检测特定的靶抗原。直接免疫荧光是两种免疫荧光类型之一。在直接免疫荧光中,单抗体(一抗)涉及,荧光团直接与原代抗体结合。在与靶抗原结合结合后,荧光团会发出荧光,可以通过荧光显微镜检测到荧光。

图01:直接免疫荧光
然而,直接免疫荧光是一种昂贵的方法,因为与二抗体相比,主要的共轭抗体成本高昂。然而,它不涉及额外的步骤,因此是一种较短的技术。此外,在直接免疫荧光中降低了非特异性结合。因此,物种交叉反应性很低。但是在检测中,与间接免疫荧光相比,直接免疫荧光的敏感性较弱。
什么是间接免疫荧光?
间接免疫荧光是第二种免疫荧光的类型,涉及两种类型的抗体,例如靶抗原标记中的初级和二抗。在这种方法中,荧光团与二抗结合。因此,该技术涉及一个额外的步骤。

Figure 02: Immunofluorescence
但是,这种方法的灵敏度很高,因为几个荧光团可以连接到二抗,并且使检测更加容易。此外,由于辅助抗体的结合更便宜且更容易,因此间接免疫荧光的价格便宜。与直接免疫荧光相比,间接方法中物种交叉反应性很高。
直接和间接免疫荧光之间的相似之处是什么?
- 它们是免疫荧光技术的两种类型。
- 初级抗体和荧光团均参与这两种方法。
- 同样,在两种方法中都出现抗原抗体反应。
- Furthermore, both direct and indirect immunofluorescence utilize a fluorescence microscope for the detection of antigen.
What is the Difference Between Direct and Indirect Immunofluorescence?
Immunofluorescence can be direct or indirect based upon the fluorophore conjugation with the antibodies used. In direct immunofluorescence, the fluorophore conjugates with the primary antibody, which is the single antibody that involves in this technique. In contrast to that, in indirect immunofluorescence, the fluorophore conjugates with the secondary antibody, which is one of the two types of antibodies that involve in the technique. Hence, the main difference between direct and indirect immunofluorescence is the antibody type that conjugates with the fluorophore.
以下信息图提供了直接免疫荧光和间接免疫荧光之间差异的并排比较。
摘要 - 直接与间接免疫荧光
Immunofluorescence is a technique that is used to detect the presence of specific antigens in a sample. This technique uses specific antibodies. Therefore, in this, the antibodies conjugate with fluorophores to detect them using a fluorescent microscope. Moreover, there are two types of immunofluorescence namely, direct and indirect immunofluorescence. Direct immunofluorescence involves a single antibody and fluorophore directly conjugated to this antibody. Indirect immunofluorescence involves two antibodies; primary and secondary and fluorophore conjugated to the secondary antibody. Since several secondary antibodies can bind with a primary antibody and several fluorophores can conjugate with secondary antibodies, indirect immunofluorescence is a more sensitive method than the direct method. Furthermore, the indirect method is less expensive than the direct method. Thus, this is the difference between direct and indirect immunofluorescence.
参考:
1.“免疫荧光。”。Wikipedia,Wikimedia基金会,2018年8月19日。在这里可用
图片提供:
1。”HSP IF IgA”By Emmanuelm at en.wikipedia,(CC由3.0)通过下议院维基梅迪亚
2. Westhayl618的“免疫荧光” - 自己的工作,(CC BY-SA 4.0)通过下议院维基梅迪亚
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