GFP4 iSIM Technical Specifications
- Andrew Vaughan
- Ki Hng
Objective lenses
| Magnification | Immersion | Type | DIC | NA | Cover slip correction | WD (mm) |
|---|---|---|---|---|---|---|
| 10x | Air | Plan Apo λ | DIC L | 0.45 | 0.17 | 4 |
| 20x | Air | Plan Apo | DIC M | 0.75 | 0.17 | 1 |
| 40x | Air | Plan Apo | DIC M | 0.95 | 0.11 - 0.23 | 0.21 |
| 60xA | Oil | Plan Apo | DIC N2 | 1.4 | 0.17 | 0.21 |
| 100x | Oil | Plan Apo λ | - | 1.45 | 0.17 | 0.13 |
| 100x†| Silicone Oil | SR HP Plan Apo λS | - | 1.35 | 0.15 - 0.19 | 0.3 |
†The 100x silicone oil lens is available on request. NA = Numerical Aperture, WD = Working Distance | ||||||
Fluorescence filter sets
Wide-field Fluorescence Filters
These filter and dichroic mirror configurations are available when the light path is oriented towards the fluorescence filter wheel and Andor Zyla camera on the right-hand port.
Quad-band filters for fast switching to the Zyla Camera. High speed but risk of cross-talk.
Filter ID | Excitation Wavelength | Dichroic Mirror | Emission Filter (in quad) | Example Fluorophores |
|---|---|---|---|---|
| DAPI | 385 nm | 86012v2 Dapi/FITC/TxRed/Cy5 | ~ 455nm - 485nm | DAPI, Hoechst |
| FITC | 470 nm | 86012v2 Dapi/FITC/TxRed/Cy5 | ~ 510nm - 545nm | GFP, FITC |
| TxRed | 550 nm | 86012v2 Dapi/FITC/TxRed/Cy5 | ~ 590nm - 630nm | RFP, mCherry |
| Cy5 | 635 nm | 86012v2 Dapi/FITC/TxRed/Cy5 | ~ 670nm - 740nm | Cy5 |
Sedat quad-band dichroic and single-band emission filters for slower switching. Lower speed but reduced cross-talk between channels.
Filter ID | Excitation Wavelength | Dichroic Mirror | Emission Filter (in wheel) | Example Fluorophores |
|---|---|---|---|---|
| DAPI | 385 nm | 89000 Sedat Quad | 435 - 485 | DAPI, Hoechst |
| GFP | 470 nm | 89000 Sedat Quad | 500 - 550 | GFP, FITC |
| RFP | 550 nm | 89000 Sedat Quad | 590 - 650 | RFP, mCherry |
| Cy5 | 660 nm | 89000 Sedat Quad | 662 - 737 | Cy5 |
| CFP | 470 nm | 89002 CFP/YFP | 460 - 500 | CFP |
| eYFP | 500 nm | 89002 CFP/YFP | 520 - 550 | YFP |
iSIM Filters
The iSIM has the following quad-band dichroic and emission filter in its scan head. The emission filter is Chroma part number 88100bs from Pinkel set 88000v2.
| Laser | Dichroic Mirror (reflection bands) | Emission Filter | Example Fluorophores |
|---|---|---|---|
| 405 | 400 - 410 | 430 - 485 | DAPI, Hoechst |
| 488 | 483 - 493 | 520 - 550 | GFP, FITC |
| 561 | 556 - 566 | 590 - 630 | RFP, mCherry |
| 640 | 635 - 645 | 680 - 740 | Cy5 |
The iSim has the following emission filters in Optospin Wheel 0
| Position | Laser | Emission band | Example Fluorophores |
|---|---|---|---|
| 1 | Any | 88000v2 Quad | Most |
| 2 | 405 | 410 - 510 | DAPI, Hoechst |
| 3 | 488 | 475 - 575 | GFP, FITC |
| 4 | 561 | 550 - 650 | tdTomato, TRITC |
| 5 | 561 | 555 - 705 | mCherry, Texas Red |
| 6 | 642 | 625 - 775 | Cy5 |
Camera Specifications
- Zyla 4.2 Plus specifications (Andor website). Note that the Zyla on GFP4 is a Camera Link model and not USB 3.0.
- Hamamatsu Orca-Flash4.0 V3 specifications (Hamamatsu website)