DeltaVision Elite Widefield Deconvolution
Image
Hardware
Our DeltaVision Elite Imaging System includes:
- Olympus iX81 stand with GEHC modifications
- Motorized XYZ stage
- Perfect Focus laser-based focus stabilization
- Solid-state fluorescence illumination with fast filter wheel
- Incubation enclosure
- PCO-edge sCMOS camera
- SoftWorx acquisition software
Filters:
Not sure which fluorophores will work best? Use an online spectra viewer (see Links) or contact Cassandra.
This system has three multiband polychroic mirrors paired with excitation and emission filters that are housed in filterwheels. The polychromatic mirros are:
- DAPI / FITC / TRITC / Cy5 = "D/F/TRITC/Cy5"
- DAPI / FITC (or GFP)/ mCherry / Cy5 = "D/F/mCherry/Cy5"
- CFP / YFP / mCherry = "C/Y/mCh"
| Software setting | Excitation | Dichroic | Emission |
|---|---|---|---|
| DAPI | 390/18nm | D/F/TRITC/Cy5 or D/F/mCherry/Cy5 |
435/48nm |
| GFP/FITC | 475/28nm | D/F/TRITC/Cy5 or D/F/mCherry/Cy5 |
525/48nm |
| TRITC | 542/27nm | D/F/TRITC/Cy5 | 597/45nm |
| mCherry | 575/25nm | D/F/mCherry/Cy5 or C/Y/mCh | 625/45nm |
| Cy5 | 632/22nm | D/F/TRITC/Cy5 or D/F/mCherry/Cy5 |
679/34nm |
| POL | White LED | na | Analyzer |
| YFP | 438/24nm | C/Y/mCh | 475/24nm |
| CFP | 513/17nm | C/Y/mCh | 548/22nm |
Objectives:
| Mag. | NA |
Immersion Medium |
DIC | Corrections | **Scaling (um/pixel) |
|---|---|---|---|---|---|
| 10x | 0.40 | Air | -- | Plan Fluor | 0.668 |
| 20x | 0.75 | Air | -- | UPlanSApo | 0.331 |
| 60x | 1.42 | Oil | -- | PlanApo N | 0.108 |
| 100x | 1.4 | Oil | -- | UPlanSApo | 0.065 |
** These values work well for making scale bars. When performing careful distance measurements, it is best practice to confirm the calibration for the exact setup you use.
Helpful Documents:
Sample holders:
All objectives expect a #1.5 glass coverslip.
| Stage plates | Compatible Sample holders |
|---|---|
| Repeatable multi-insert stage | Slides, LabTek II chambers, 35mm dishes |
| Stage ring | Other formats, including portions of multiwell plates |
Software:
To view files in lab, many users like to use Fiji (ImageJ) with the BioFormats plugin for importing the .dv file. See our Links page to download.