Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Steps 1–4 | |||
| Concavity slides | ThermoFisher Scientific | 1519006 | 5–18mm diameter, 0.6–0.8mm deep, 1.2–1.5mm |
| Dissecting microscope with 10×– 50× zoom range | Motic | SMZ-171 | |
| E. coli (OP50) | Caenorhabditis Genetics Center (CGC) | OP50 | |
| Glass coverslips, no. 1.5, 24 × 50 mm | VWR International | 48393–241 | |
| M9 Buffer | Stiernagle, T. Maintenance of C. elegans. WormBook. 1–11, doi:10.1895/wormbook.1.101.1, (2006). | ||
| Methyl cellulose | Sigma-Aldrich | H7509–25G | |
| Microcapillary pipette aspirator tube | Sigma-Aldrich | A5177 | |
| Microcapillary pipettes, 0.4-mm i.d | Drummond Scientific | 1–000-800 | |
| Needle, no. 18G x 1 ½ (1.2mm x 40mm) | BD Precision Glide | 305196 | |
| NGM plates | prepared as described by Brenner (1974) | ||
| O-ring for imaging chamber | O-Rings West | M1.5X40 | |
| Pasteur pipette | Corning/Sigma-Aldrich | CLS7095D5X | |
| Platinum wire, 0.5-mm diameter | Sigma-Aldrich | 267201 | |
| Poly-L-lysine | Sigma-Aldrich | P1524 | |
| Stainless steel rectangular chamber (76.0 mm x 50.5 mm) | Applied Scientifics Instrumentations (ASI) | I2450 | |
| Worm Eyelash Pick | Hart, A. C. Behavior.
WormBook. (2006). |
||
| Worm Pick | Stiernagle, T. Maintenance of C. elegans. WormBook. 1–11, doi:10.1895/wormbook.1.101.1, (2006). |
||
| Name | Company | Catalog Number | Comments |
| Steps 5–6 | |||
| 488 nm long-pass filter | Semrock | LP02–488 RU-2 | |
| 561-nm notch filter | Semrock | NF03–561E-25 | |
| BLP02–561R-25, quantity 2 | Semrock | 561 nm EdgeBasic best-value long-pass edge filter | |
| Control software for bottom camera | Jenoptik | ProgRes CapturePro | |
| diSIPM assembly video | Applied Scientifics Instrumentations (ASI) | https://youtu.be/TAgbr6IrTqw ; http://www.asiimaging.com | |
| diSPIM alignment video | Applied Scientifics Instrumentations (ASI) | https://youtu.be/qnOrg30NNuE | |
| diSPIM imaging PC | Intel | Intel Xeon CPU E5–2630 2.6GHz, 12 cores in total, 64 GB memory, Windows 7 | |
| FF01–525/45–25, quantity 2 | Semrock | 525/45 nm BrightLine single-band bandpass filter | |
| FF555-DI03–25X36, quantity 2 | Semrock | 555 nm edge BrightLine single- edge dichroic beamsplitter | |
| Imaging PC Graphics Card | NVIDIDA | NVIDIA GeForce GTX 1080 Ti graphics cards |
|
| Kumar et al diSPIM Setup | Applied Scientifics Instrumentations (ASI) | Instrument setup for this protocol is identical to Kumar et al 10,11 diSPIM, which makes use of 40× 0.8NA water immersion lenses for imaging. (See steps 5.1 and note) |
|
| Micro Manager | Micro-Manager | https://micro-manager.org/ | |
| Modifications to Kumar et al diSPIM Setup (see below) | |||
| Optical table with isolators, 4 feet × 6 feet × 12 inches | TMC | 784–651-02DR and 14–416-34 | |
| Name | Company | Catalog Number | Comments |
| Steps 7–10 | |||
| Analysis PC | Intel | Intel Core i7–8700K CPU 3.70GHz, 6 cores in total, 64 GB memory, Windows 10 | |
| Analysis PC Graphics Card | NVIDIDA | NVIDIA GeForce GTX 1080 Ti graphics cards |
|
| Installation instructions | Software bundle | http://dispimlineage.wormguides.org/ diSPIMlineaging_InstallationInstructi | |
| Software bundle | Software bundle | http://dispimlineage.wormguides.org |