Nikon AXR/NSPARC + STED
The Nikon AXR is a point scanning confocal built around the inverted Ti2E microscope, with additional resolution provided by the NSPARC APD array detector, which improves signal-to-noise and extends resolution, and a super resolution Abberior STEDYCON module, with lateral resolution down to 30nm.
- Nikon inverted Ti2E with PerfectFocus gen. 4
- Prior piezo z stage with 600µm range
- Okolab stagetop incubator
Laser units
- LUA-S4 and LunF-combiner 405 / 445 / 488 / 514 / 561 / 594 / 640 / 730
Dichroic mirrors
- Standard quad (405/488/561/640)
- Standard penta (405/488/561/640/730)
- "FRET" penta (405/445/514/594/730)
Detector units
- DUX with 3 GaAsP (2 tunable) and 1 IR-specific GaAsP PMT detectors
- NSPARC array detector of 25 APDs with down to 100 nm lateral x 300 nm axial resolution
| Mag | NA | W.D. (mm) | Immersion | Corr |
|---|---|---|---|---|
| 2X* | 0.1 | 8.5 | Air | Plan Apo Lambda D |
| 10X | 0.45 | 4 | Air | Plan Apo Lambda D |
| 20X* | 0.8 | 0.8 | Air | Plan Apo Lambda D |
| 20X | 0.95 | 0.99 - 0.9 | WI | Apo LWD Lambda S |
| 25X | 1.05 | 0.55 | Sil | Plan Apo Lambda S |
| 40X | 1.15 | 0.61 - 0.59 | WI | Apo LWD Lambda S |
| 60X | 1.42 | 0.15 | Oil | Plan Apo Lambda D |
| 100X | 1.45 | 0.13 | Oil | Plan Apo Lambda D |
*Available to be put in instrument, please contact Emery to request use. Use the 100X for optimal STED.
NSPARC
The NSPARC array detector is composed of 25 individual APDs (avalanche photodiodes), which are photon counters and provide higher signal-to-noise than standard detectors. The NSPARC is capable of lateral resolution down to 100nm.
NIS Elements confocal package is used for acquisition. The offline Nikon workstation in Steger Hall also includes General Analysis 3, batch denoise, and batch 2D/3D deconvolution tools.
Abberior STED module
The STEDYCON unit is a convenient, shoebox version of Abberior's STED instruments. It is capable of acquiring up to 3 STED channels with one STED laser, given a long Stoke's shift green (488 nm excitation) dye is used, improving lateral resolution down to 30 nm. STED can be performed on a variety of fixed and live samples using appropriate dyes. A table of verified STED dyes is shown below.
STEDYCON with STEDYFOCUS
- Alignment-free design
- Excitation lasers: 405 (CW); 488 / 561 / 640 nm pulsed
- STED laser: 775 nm pulsed
- Scanner: QUAD beam, 90µm x 80µm FOV (at 100X) with speed up to 1 FPS
- Detectors: time-gated APDs
- Browser-based software that runs on module for uninterrupted imaging
A 2D view of a diffraction-limited excitation point (blue) overlapped by a STED donut (orange) is shown on the upper left, with a graphical representation of the gaussian overlap below. The effective fluorescent spot and graph are shown on the right in green.
