Table 2 Application, advantages, and disadvantages of microscopic techniques with antibody-based probes in AGPs studies
Technique | Application | Advantage | Disadvantage |
|---|---|---|---|
Immunofluorescence labelling—Confocal laser scanning microscope (CLSM) | Used to show where AGPs are distributed across labelled cells and tissues; analyses at the tissue/cellular level | AGP location is identified using a fluorescent signal It is possible to see cellular features at a resolution of 1 µm High contrast and resolution images are possible to obtain quickly and non-intrusively Eliminates the problem of glare resulting from preparation of layers lying outside the plane of focus | The fluorescence lessens with time. The fluorescence of samples fades (photobleaching) during observations Results are susceptible to the effects of the environment Resolution is still inferior to that of electron microscopy A high price and a very limited field of view |
Immunogold labelling—Transmission electron microscope (TEM) | Used to localise AGPs and other cell wall components at the subcellular level | AGP localisation in ultra-thin sections using a gold-conjugated antibody It is possible to see intracellular features at a resolution of 0.2 nm Analysis and observation of very high-resolution images High magnification | Fixation is needed in cell preparation, which might result in artificial damage Lengthy and complicated methods for preparation of cells and tissues for TEM Requires a conductive coating of gold/palladium alloy, carbon, osmium, etc. for correct imaging Expensive, difficult to access, and complicated to use |