Traditional immunofluorescence techniques are mature and reliable, easy to operate, yield stable results, and have lower costs. In comparison, TSA has its unique advantages: high sensitivity, the ability to label multiple proteins, and no restrictions on antibody species, thus compensating for the shortcomings of traditional immunofluorescence. In specific experiments, if only routine detection is required, traditional immunofluorescence is sufficient; however, for in-depth investigation of fine structures, TSA is indispensable.
The table below compares several methods：
|
Technology |
Immunohistochemical (IHC) |
Immunofluorescence (IF) |
TSA mIHC |
|
Color rendering method |
DAB simple staining DAB+ red color original bichrome |
Primary and secondary staining was mixed |
Multiple rounds of repeated fluorescence experiments |
|
Type of primary antibody |
compliant IHC |
The species of the primary antibody should be different for multiple indicators |
The species of multi-antibody is not limited |
|
Type of secondary antibody |
HRP-labeled secondary antibody |
Different species of secondary antibody labeled with different fluorophores |
HRP-labeled secondary antibody |
|
Coloration effect |
Only two indicators can be stained at the same time; Double staining cannot observe the co-expression, and the two indicators need to be located differently; Indicators cannot be observed in a single channel during double staining |
The detection sensitivity is low It is easy to quench At the same time, the number of eggs marked is small
|
The detection sensitivity is high and the fluorescence signal is doubled Strong anti-quenching Multiple proteins (up to 7-10) are also tagged |
|
Detection costs |
If you need to stain multiple indicators, you need to cut many slices, which is not suitable for precious or small sample size samples |
The species of antibody should be different, and the selection range is limited Mixed staining is not conducive to exploring experimental conditions and increasing experimental costs |
The species of antibody is not limited, and the selection range is wide The experimental conditions can be adjusted at any time in the pre-experiment of multi-round staining |
