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start [2018/07/22 13:56]
Jon Daniels [The Implementation] |
start [2018/09/26 20:51]
Jon Daniels [The Implementation] |
| The diSPIM "head" can be mounted on various inverted microscopes including ASI's RAMM frame (shown here). diSPIM systems can be obtained from various [[docs:system integrators]]. Various open-source and proprietary software packages are available for both data acquisition and data processing. Most of the underlying microscope hardware is identical regardless of the system integrator and software used. | The diSPIM "head" can be mounted on various inverted microscopes including ASI's RAMM frame (shown here). diSPIM systems can be obtained from various [[docs:system integrators]]. Various open-source and proprietary software packages are available for both data acquisition and data processing. Most of the underlying microscope hardware is identical regardless of the system integrator and software used. |
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| The choice of diSPIM objectives is limited because they must be co-focused without bumping into each other. The most commonly-used objectives for diSPIM are 40x water-dipping objectives with a NA of 0.8 (Nikon CFI Apo 40XW NIR). The Olympus 20x/0.5 objective is another possibility((According to the manufacturer's drawings the objectives co-focus exactly when they touch, which also implies zero gap between objective and coverslip. However, in practice they co-focus with a gap comparable to the 40x/0.8, with ~200um between the objective bottom and coverslip.)) as is the Nikon 10x/0.3. ASI and Special Optics have co-developed an [[http://asiimaging.com/docs/cleared_tissue_objective|custom objective for cleared tissue]] that is suitable for the diSPIM and can image cleared tissue up to 5 mm deep in slab form or within a 12 mm spherical envelope. Single-sided systems (iSPIM) have much more flexibility because the illumination objective can be a low-NA long-WD objective. | The choice of diSPIM objectives is limited because they must be co-focused without bumping into each other. The most commonly-used objectives for diSPIM are 40x water-dipping objectives with a NA of 0.8 (Nikon CFI Apo 40XW NIR). The Olympus 20x/0.5 objective is another possibility((According to the manufacturer's drawings the objectives co-focus exactly when they touch, which also implies zero gap between objective and coverslip. However, in practice they co-focus with a gap comparable to the 40x/0.8, with ~200um between the objective bottom and coverslip.)) as is the Nikon 10x/0.3. ASI and Special Optics have co-developed an [[http://asiimaging.com/docs/cleared_tissue_objective|objective for cleared tissue]] that is suitable for the diSPIM and can image cleared tissue up to 5 mm deep in slab form or within a 12 mm spherical envelope. Single-sided systems (iSPIM) have much more flexibility because the illumination objective can be a low-NA long-WD objective. |
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| Most often sCMOS cameras are used for SPIM imaging. There are working diSPIM systems with Hamamatsu Flash4, Andor Zyla, PCO Edge, and Photometrics Prime 95B cameras. | Most often sCMOS cameras are used for SPIM imaging. There are working diSPIM systems with Hamamatsu Flash4, Andor Zyla, PCO Edge, and Photometrics Prime 95B cameras. |
