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Both sides previous revision Previous revision Next revision | Previous revision Last revision Both sides next revision | ||
docs:modular_manual [2019/02/27 23:08] Melissa Glidewell [Orientation] |
docs:modular_manual [2022/08/25 07:03] Jon Daniels [Filters and Mirrors] |
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===== Mount the SPIM Head ===== | ===== Mount the SPIM Head ===== | ||
{{ youtube> | {{ youtube> | ||
- | **Tools**: | + | **Tools**: |
The video is cued up to show the process for mounting a diSPIM onto an ASI RAMM frame, but the principles apply to other scopes as well.\\ | The video is cued up to show the process for mounting a diSPIM onto an ASI RAMM frame, but the principles apply to other scopes as well.\\ | ||
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===== Red Bracket ===== | ===== Red Bracket ===== | ||
- | **Tools**: | + | **Tools**: |
RAMM frames are shipped with a red bracket supporting the inverted scope; it must be removed. For detailed instructions with pictures, please visit [[http:// | RAMM frames are shipped with a red bracket supporting the inverted scope; it must be removed. For detailed instructions with pictures, please visit [[http:// | ||
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===== Scanners ===== | ===== Scanners ===== | ||
{{ youtube> | {{ youtube> | ||
- | **Tools**: | + | **Tools**: |
The video is cued up to show the process for mounting the scanners.\\ | The video is cued up to show the process for mounting the scanners.\\ | ||
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===== Orientation ===== | ===== Orientation ===== | ||
{{ youtube> | {{ youtube> | ||
- | **Tools**: | + | **Tools**: |
The video is cued up to show the process for mounting and orienting the cameras.\\ | The video is cued up to show the process for mounting and orienting the cameras.\\ | ||
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===== Filters and Mirrors ===== | ===== Filters and Mirrors ===== | ||
{{ youtube> | {{ youtube> | ||
- | **Tools**: ASI ring tool, small Phillips head screwdriver, | + | **Tools**: ASI ring tool, small Phillips head screwdriver, |
The video is cued up to show the process for installing an excitation filter, emission filter, and dichroic filter.\\ | The video is cued up to show the process for installing an excitation filter, emission filter, and dichroic filter.\\ | ||
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<WRAP left round info 50%> | <WRAP left round info 50%> | ||
- | Notice: It is very easy to warp dichroic mirrors; only lightly tighten the retaining clip of the D-CUBE. | + | Notice: It is easy to warp dichroic mirrors |
</ | </ | ||
\\ \\ \\ \\ \\ | \\ \\ \\ \\ \\ | ||
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===== Stage limits ===== | ===== Stage limits ===== | ||
- | **Tools**: | + | **Tools**: |
Setting the limit magnets on the motorized stages prevents crashes that can break coverslips or otherwise damage equipment; hardware limits are the most foolproof way to prevent crashes.((Limits may also be defined in software. Software-defined limits persist when the controller is powered down, but are lost when the firmware is updated or reset.)) The limit magnets move with the stage plates; when a magnet approaches a Hall effect sensor affixed in the stage body, the firmware detects a limit condition and stops the stage. The polarity of limit magnets relative to the stage is essential to their function. | Setting the limit magnets on the motorized stages prevents crashes that can break coverslips or otherwise damage equipment; hardware limits are the most foolproof way to prevent crashes.((Limits may also be defined in software. Software-defined limits persist when the controller is powered down, but are lost when the firmware is updated or reset.)) The limit magnets move with the stage plates; when a magnet approaches a Hall effect sensor affixed in the stage body, the firmware detects a limit condition and stops the stage. The polarity of limit magnets relative to the stage is essential to their function. | ||
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===== Single axis ===== | ===== Single axis ===== | ||
- | {{ youtube> | + | {{ youtube> |
The video is cued up to demonstrate setting the F-axis limit magnet.\\ | The video is cued up to demonstrate setting the F-axis limit magnet.\\ | ||
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===== XY-axes ===== | ===== XY-axes ===== | ||
{{ youtube> | {{ youtube> | ||
- | **Tools**: | + | **Tools**: |
The video is cued up to demonstrate setting the XY-axes limit magnets; note that we now recommend setting the X-axis magnets first.\\ | The video is cued up to demonstrate setting the XY-axes limit magnets; note that we now recommend setting the X-axis magnets first.\\ | ||
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===== Beam position ===== | ===== Beam position ===== | ||
- | We are concerned with getting both the beam position and angle correct. Fortunately, | + | We are concerned with getting both the beam position and angle correct, along with getting both objectives " |
< | < | ||
^ Adjustment | ^ Adjustment | ||
- | | Tilt of dichroic mirror | + | | Tilt of dichroic mirror |
- | | Tilt of camera mirror | + | | Tilt of camera mirror |
- | | Position of upper Z stage | No | No | Angle varies if scanners are not connected to dichroic tubes | + | | Z position of SPIM head | No | No | |
| Objective bushing (focus) | | Objective bushing (focus) | ||
| Linear objective adjustment | | Linear objective adjustment | ||
- | | < | + | | < |
- | | < | + | | < |
</ | </ | ||
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==== Phase 1 ==== | ==== Phase 1 ==== | ||
- | Repeat the steps of Phase 1 steps multiple times (after the first iteration, in no particular order) | + | Repeat the steps of Phase 1 steps multiple times as you spiral in on alignment. |
=== Cofocus SPIM objectives === | === Cofocus SPIM objectives === | ||
{{ youtube> | {{ youtube> | ||
- | **Adjust**: OBLPA (<imgref piezo_with_bushing> | + | **Configure** the [[docs: |
- | **End goal**: Both beams in focus with the epi-spot sitting on the beam waist (roughly at first, as in <imgref BeginAlign>, | + | select |
- | The video is cued up to demonstrate cofocusing | + | use the "On tab activate" |
- | In the [[docs:mm_dispim_plugin_user_guide# | + | **Adjust**: |
+ | * both objective bushings, and | ||
+ | * the knob on the OBLPA (<imgref piezo_with_bushing> | ||
+ | **The goal** is to have both beams in focus (roughly at first, as in <imgref BeginAlign>, | ||
+ | The video is cued up to demonstrate cofocusing the SPIM objectives using Micro-Manager.\\ | ||
This is best done by alternating between steering and focus adjustments, | This is best done by alternating between steering and focus adjustments, | ||
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**Adjust**: Rotation of the camera (and camera' | **Adjust**: Rotation of the camera (and camera' | ||
**End goal**: Epi-view of the sheets produces a vertical line in the camera image\\ | **End goal**: Epi-view of the sheets produces a vertical line in the camera image\\ | ||
- | **Tools**: | + | **Tools**: |
The video is cued up to demonstrate adjustment of the camera' | The video is cued up to demonstrate adjustment of the camera' | ||
- | If the epi-view of the sheet is not vertical, | + | If the epi-view of the sheet is not vertical, |
If you make any changes in Phase 2, return to the start of Phase 1. | If you make any changes in Phase 2, return to the start of Phase 1. |