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docs:manual [2019/01/21 18:18] Melissa Glidewell updated imgref |
docs:manual [2019/02/27 23:02] Melissa Glidewell [Mount] |
====== Overview ====== | ====== Overview ====== |
{{section>modular_manual#overview&noheader}} | {{section>modular_manual#overview&noheader}} |
The advantage of the diSPIM being dual-sided is that the role of the two objectives can be reversed to collect a second stack from a perpendicular direction. The two optical paths are usually symmetric, and are designated Path A and Path B. (The two paths are described in detail in the [[docs:mm_dispim_plugin_user_guide#diSPIM Plugin User Guide]].) Registration and joint deconvolution merge the two datasets for improved resolution isotropy. | |
| The core assembly, the SPIM head, can be mounted on various inverted microscopes. Thus far, microscope-specific adapter brackets((Attaching the SPIM precludes the use of the transmitted light optics because the brackets utilize the transmitted light pillar mounting point.)) exist for the following microscopes: |
| * ASI RAMM frame (<imgref SPIM_Figure>) |
| * Leica DMI-6000 |
| * Nikon TE-300, Ti, TE-2000, Ti2 |
| * Olympus IX-71/81, IX-73/83 |
| * Zeiss Axio-Observer |
| |
<imgcaption SPIM_Figure|Typical complete diSPIM system on RAMM frame showing major components. 1) SPIM-MOUNT , 2) SPIM-RAMM , 3) SPIM arm mount (RAO-0046 ), 4) piezo objective mover APZOBJ-200 or similar, 5) MIM inverted microscope with LS-50M stage, 6) CDZ-1000 centering stage or CDZ-R block, 7) LS-50M stage, 8) MIM-CUBE-II w/ mirror, 9) MIM-CUBE-II w/ fluorescence filters, 10) C60-TUBE_B imaging tube lens, 11) C60-TUBE_160 scanner tube lens, 12) 50mm extension tube, 13) MM-SCAN_1M light sheet scanner, 14) MS-2500 XY stage.>{{:docs:manual:dispim_figure.jpg?600| Typical complete diSPIM system on RAMM frame showing major components.}}</imgcaption> | <imgcaption SPIM_Figure|Typical complete diSPIM system on RAMM frame showing major components. 1) SPIM-MOUNT , 2) SPIM-RAMM , 3) SPIM arm mount (RAO-0046 ), 4) piezo objective mover APZOBJ-200 or similar, 5) MIM inverted microscope with LS-50M stage, 6) CDZ-1000 centering stage or CDZ-R block, 7) LS-50M stage, 8) MIM-CUBE-II w/ mirror, 9) MIM-CUBE-II w/ fluorescence filters, 10) C60-TUBE_B imaging tube lens, 11) C60-TUBE_160 scanner tube lens, 12) 50mm extension tube, 13) MM-SCAN_1M light sheet scanner, 14) MS-2500 XY stage.>{{:docs:manual:dispim_figure.jpg?600| Typical complete diSPIM system on RAMM frame showing major components.}}</imgcaption> |
The relevant sections of the diSPIM [[https://youtu.be/TAgbr6IrTqw|assembly]] video are embedded. | The relevant sections of the diSPIM [[https://youtu.be/TAgbr6IrTqw|assembly]] video are embedded. |
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===== Mount the SPIM Head ===== | ===== SPIM Head ===== |
{{section>modular_manual#mount_the_spim_head&noheader}} | {{section>modular_manual#mount_the_spim_head&noheader}} |
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\\ | \\ |
<imgcaption SPIM_mount_to_RAMM| SPIM head mounts via 2 dowel pins and a bolt at the indicated location regardless of the microscope frame. (RAMM frame pictured)>{{:docs:manual/SPIM_mount_to_RAMM.png| The SPIM head mounted to the microscope frame via 2 dowel pins and a bolt.}}</imgcaption> | <imgcaption SPIM_mount_to_RAMM| SPIM head mounts via 2 dowel pins and a bolt at the indicated location regardless of the microscope frame. (RAMM frame pictured)>{{:docs:manual/SPIM_mount_to_RAMM.png| The SPIM head mounted to the microscope frame via 2 dowel pins and a bolt.}}</imgcaption> |
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| ===== Red Shipping Bracket ===== |
| {{section>modular_manual#red_bracket&noheader}} |
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===== Scanners ===== | ===== Scanners ===== |
{{section>modular_manual#camera_setup&noheader}} | {{section>modular_manual#camera_setup&noheader}} |
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==== Mount ==== | ==== Mount Camera Tube Lenses ==== |
Tools: M5 hex driver\\ | Tools: M5 hex driver\\ |
In the standard configuration for diSPIM on a RAMM frame, as in <imgref SPIM_Figure>, each camera tube lens is fastened to the SPIM-MOUNT((Other common configurations use a post mount system to minimize vibration from the camera's fans, or to accommodate an FTP system in lieu of a RAMM frame.)) with a ring clamp. The end of the tubes should come within approximately 1 mm from the upper (mirror) CUBE-IIIs. When the objectives are lowered into position for viewing a sample, adjust the bolts holding the tube support arms so the camera tube lenses are centered on the CUBE-IIIs. \\ | In the standard configuration for diSPIM on a RAMM frame, as in <imgref SPIM_Figure>, each camera tube lens is fastened to the SPIM-MOUNT((Other common configurations use a post mount system to minimize vibration from the camera's fans, or to accommodate an FTP system in lieu of a RAMM frame.)) with a ring clamp. The end of the tubes should come within approximately 1 mm from the upper (mirror) CUBE-IIIs. When the objectives are lowered into position for viewing a sample, adjust the bolts holding the tube support arms so the camera tube lenses are centered on the CUBE-IIIs. \\ |
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===== Optical alignment ===== | ===== Optical alignment ===== |
| |
| **Goals:** |
| The qualities of final alignment, for both SPIM optical paths, include all of the following: |
| * epi spot centered on epi camera |
| * beam waist centered on imaging camera |
| * beam/sheet plane coincident with imaging focus plane near the center of the piezo travel |
| * beam perpendicular to imaging camera field of view |
| * sheet centered in imaging camera field of view |
| * sheets centered in bottom camera’s field of view (i.e. co-alignment of all three objectives) |
| |
==== Overview ==== | ==== Overview ==== |
{{section>modular_manual#alignment_overview&noheader}} | {{section>modular_manual#alignment_overview&noheader}} |
Goals | |
The qualities of final alignment, for both SPIM optical paths, include all of the following: | |
epi spot centered on epi camera | |
beam waist centered on imaging camera | |
beam/sheet plane coincident with imaging focus plane near the center of the piezo travel | |
beam perpendicular to imaging camera field of view | |
sheet centered in imaging camera field of view | |
sheets centered in bottom camera’s field of view (i.e. co-alignment of all three objectives) | |
Exploring beam position and angle: what affects what? | |
{{section>modular_manual#beam_position&noheader}} | {{section>modular_manual#beam_position&noheader}} |
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