What are the Common Types of Multiplex Staining Techniques?
Several techniques are employed for multiplex staining, each with its unique advantages and limitations:
1. Immunofluorescence (IF) Multiplexing: This method uses multiple fluorescently labeled antibodies, each targeting a different antigen. The use of distinct fluorophores allows for the simultaneous detection of multiple proteins.
2. Chromogenic Multiplexing: Utilizes enzyme-labeled antibodies to produce colored precipitates. Different enzymes or substrates produce different colors, enabling the detection of multiple targets.
3. Mass Cytometry (CyTOF): Combines flow cytometry with mass spectrometry to label antibodies with rare metal isotopes. This technique allows for the simultaneous detection of over 40 markers without spectral overlap.
4. Tyramide Signal Amplification (TSA): Employs enzyme-conjugated antibodies and tyramide signal amplification to enhance the signal of each target. Sequential rounds of staining and stripping can be used to visualize multiple markers.
