Table 2 |.
Examples of compatible microscope excitation and emission configurations
| Example system with a broadband light source | Example system with a multispectral laser light source, external filter wheel, and triggered acquisition | Notes | |
|---|---|---|---|
| Approximate acquisition time for one field-of-view of imaging with DAPI and SBS channels | 5300 ms | 610 ms | |
| Light source | Sola SE FISH 2 light engine (Lumencor) | CELESTA or CELESTA quattro light engine (Lumencor) | The Sola is more economical but has reduced power, requiring longer exposures especially for base C. CELESTA is available with different laser line configurations; only 405 nm, 545 nm, and 635 nm are strictly needed for sequencing. |
| External filter wheel | N/A | HS-625 (Finger Lakes Instrumentation) or Lambda 10-B (Sutter Instrument) | Enables fast emission filter switching |
| Components for external hardware triggering | N/A | NIS-Elements Hardware Triggering module, compatible NI-DAQ card |
Reduces communication overhead in synchronizing components |
| Multiband dichroic mirror | N/A | ZT408/473/545/635/750rpc (Chroma) or Di01-R488/543/635 (Semrock) |
When using an external emission filter wheel, a multiband dichroic mirror is required in the microscope body filter turret. The first option works for all suggested SBS and phenotype channels but is expensive. The second option is more economical but does not work for DAPI or near-infrared channels. |
| DAPI filter set | LED-DAPI-A filter set (Semrock) | 405 nm laser line excitation | |
| ZET408/473/545/635/750m (Chroma) or any compatible single-band emission filter | |||
| GFP filter set | GFP-1828A filter set (Semrock) | 473 nm laser line excitation | For phenotype imaging |
| Same multiband filter as for DAPI or any compatible single-band emission filter | |||
| MiSeq G filter set | FF01–534/20 (Semrock) | 545 nm laser line excitation | |
| FF552-Di02 (Semrock) | |||
| FF01–572/28 (Semrock) | FF01–565/24 (Semrock) | ||
| MiSeq T filter set | FF03–575/25 (Semrock) | 545 nm laser line excitation | |
| FF596-Di01 (Semrock) | |||
| FF01–615/24 (Semrock) | FF01–624/40 (Semrock) | ||
| MiSeq A filter set | FF01–635/18 (Semrock) | 635 nm laser line excitation | |
| FF652-Di01 (Semrock) | |||
| FF01–680/42 (Semrock) | FF01–676/29 (Semrock) | ||
| MiSeq C filter set | FF01–661/20 (Semrock) | 635 nm laser line excitation | |
| FF695-Di01 (Semrock) | |||
| FF01–732/68 (Semrock) | FF01–732/68 (Semrock) | ||
| near-IR filter set | N/A | 750 nm laser line excitation | Optional for extending phenotype fluorophore options (e.g., Alexa Fluor 750). |
| FF765-Di01 (Semrock) | |||
| ET820/110 (Chroma custom) |