===== Comparison of diSPIM With Confocal ===== The following comparison was written with diSPIM in mind, but the main points apply to all types of SPIM/LSFM. Spinning disk confocal (SDCM) is just a massively parallel confocal implementation so it's faster but otherwise has the same characteristics as laser scanning confocal. * **Light dose:** diSPIM has much lower dose and hence less photobleaching/phototoxicity than confocal. Reducing light dose is the main motivation for most SPIM users. * A rule of thumb is that you need X times as much light for confocal as you do for SPIM where X is the number of slices in your stack. * [[http://www.nature.com/nbt/journal/v31/n11/fig_tab/nbt.2713_F3.html|Figure 3]] and [[http://www.nature.com/nbt/journal/v31/n11/fig_tab/nbt.2713_SV2.html|Supplementary Video 2]] of Wu et al. offer bleaching comparisons of imaging with diSPIM and SDCM. Other papers such as [[http://dx.doi.org/10.1002/jbio.201200144|this one]] offer comparisons of light sheet with confocal which should apply to diSPIM. * **Z resolution:** diSPIM after registration/fusion has best Z-resolution (same as XY resolution). Next best is confocal, which in turn has better Z resolution than single-view SPIM. * Using Bessel beams to create the sheet can improve Z-resolution for single-view SPIM ("optical sectioning"), but won't meaningfully change dual view resolution after registration/fusion. * **XY resolution:** confocal and SPIM are comparable. * **Speed:** SPIM (per view) and SDCM have comparable speed if SDCM laser intensity is increased to compensate for the ~3% open area of the disks. * Camera readout speed bounds the maximum achievable frame rate. For example, 512 pixels high ROI is 2.5 ms readout time for sCMOS. Allowing 2.5 ms illumination time results in 5 ms total per image or 200 frames per second. Using a cylindrical lens instead of a scanned light sheet can reduce camera overhead.