Amyotrophic lateral sclerosis (ALS or Lou Gehrig’s disease) is a neurodegenerative disorder that is characterized by the progressive loss of motor neurons. Consequently, patients afflicted with this disease eventually lose the motor function associated with moving, speaking, eating, and even breathing. One neuroscience team identified a common mechanism in nearly all sporadic ALS patients in which transactive response DNA binding protein 43 kDa (TDP-43) was mislocalized from the nucleus to the cytoplasm. This suggests that the mislocalization of TDP-43 into the cytoplasm is central for the onset of ALS in patients, but the underlying factors that result in this process are not fully understood as there are no current laboratory methods to study the progression of TDP-43 mislocalization. However, the development and refinement of lattice light-sheet microscopy (LLSM) as a laboratory technique has significant potential in helping scientists elucidate the unknown processes behind ALS. This review discusses the implications of lattice light-sheet microscopy and its optimization methods as a solution to the issue and identifies LLSM as a key method that could be significant in the field of ALS research.