Modification and improvement of immunohistochemical double staining for tumor associated macrophages in lung adenocarcinoma tissues

Xianchao Zhang; Xiang Zhang; Mianfu Cao; Lu Chen; Weiqi Dang; Kang Song; Youhong Cui; Xia Zhang; Xiuwu Bian

doi:10.16016/j.1000-5404.201701070

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Modification and improvement of immunohistochemical double staining for tumor associated macrophages in lung adenocarcinoma tissues

更新时间：2026-01-09

- Modification and improvement of immunohistochemical double staining for tumor associated macrophages in lung adenocarcinoma tissues
- Journal of Third Military Medical University Vol. 39, Issue 11, Pages: 1155-1158(2017)
- 作者机构：
- 作者简介：
- 基金信息：
  
  the National Key Research and Development Program(2016YFC201801);the General Program of National Natural Science Foundation of China(81372273);the Science and Technology Innovation Project for Intensifying Hospital of Southwest Hospital(SWH2016JCZD-04)
- DOI：10.16016/j.1000-5404.201701070
  CLC：
- Received：11 January 2017，
  
  Published：15 June 2017
- 稿件说明：
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Xianchao Zhang, Xiang Zhang, Mianfu Cao, et al. Modification and improvement of immunohistochemical double staining for tumor associated macrophages in lung adenocarcinoma tissues[J]. Journal of Third Military Medical University, 2017, 39(11): 1155-1158.
DOI：

Xianchao Zhang, Xiang Zhang, Mianfu Cao, et al. Modification and improvement of immunohistochemical double staining for tumor associated macrophages in lung adenocarcinoma tissues[J]. Journal of Third Military Medical University, 2017, 39(11): 1155-1158. DOI： 10.16016/j.1000-5404.201701070.

摘要

Abstract

Objective

To explore a stable and easy-to-identify color developing method by modifying immunohistochemical double-staining (D-IHC) so as to investigate the distribution of tumor associated macrophages (TAMs) in lung adenocarcinoma tissue.

Methods

Although as the same as the traditional double-enzyme methods which used the enzyme-conjugated secondary antibody to respectively link alkaline phosphatase (ALP) and horseradish peroxidase (HRP) as catalyze chromogenic substrates

AP-Red and diaminobenzidine (DAB) were chosen instead of 5-Bromo-4-Chloro-3-Indolyl Phosphate (BCIP/NBT) and 3-Amino-9-ethylcarbazole (AEC) as chromogenic substrates. The distribution of TAMs in lung cancer tissues was observed under the microscope after the double staining were controlled in order.

Results

As the image was highly contrast and easily distinguishable with low background under a light microscope

Fast Red (first labeling) and DAB (second labeling) were suitable for application in D-IHC. In addition

these 2 kinds of color labeling were merged as brick red at the same position

which was recognizable under a light microscope. The statistical results revealed that M2 type of TAMs were priority (accounting for 80%) in the lung adenocarcinoma tissue samples.

Conclusion

The modified method is simple and reliable with stable labeling results

and will be useful in the exploration of lung adenocarcinoma and other biomedical research.

关键词

Keywords

references

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