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docs:comparison_with_confocal [2016/01/26 17:51]
Jon Daniels
docs:comparison_with_confocal [2016/05/09 23:18] (current)
Jon Daniels
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 The following comparison was written with diSPIM in mind, but the main points apply to all types of SPIM/LSFM. 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 some characteristics as laser scanning confocal.+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.   * **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.
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       * [[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.       * [[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.   * **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, but won't meaningfully change dual view resolution after registration/fusion.+      * 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.   * **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.   * **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.+      * 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.