The advantages of the method include:
* uniquely wide field of view (because translation complexes of any type, including scanning small ribosomal subunits, are captured for the first time);
* potentially more natural representation of complex dynamics (because all, and not only selected, translation processes are arrested by formaldehyde fixation);
* possibly more faithful and/or sensitive detection of translation complexes locations (as covalent fixation prevents detachment of the fragments from the ribosomes or their subunits).
The disadvantages include:
* higher overall complexity of the experimental procedure (due to requirement of the initial isolation of translated mRNA and preparative sedimentation to separate ribosomes and ribosomal subunits);
* higher contamination of the useful sequencing read depth with the undesired fragments of the ribosomal RNA (inherited from the wide size selection window used for protected RNA fragments);
* a pre-requirement for optimization of the formaldehyde fixation procedure for each new cell or sample type (as optimal formaldehyde fixation timings strongly depend on sample morphology and both over- and under-fixation will compromise the results).