Nikon AXR with NSPARC and STEDYCON
Overview
The Nikon AXR is a point scanning confocal built around the inverted Ti2E, with additional resolution provided by the NSPARC detector which improves signal-to-noise, and a super resolution Abberior STEDYCON module, with lateral resolution down to 30nm.
Specifications
Microscope
- 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 / 515 / 561 / 594 / 640 / 730
Objective Lenses
- 2x/0.1 Plan Apochromat Lambda D*
- 10x/0.45 Plan Apochromat Lambda D
- 20x/0.8 Plan Apochromat Lambda D*
- 20X/0.95 Apochromat LWD Lambda S, WI
- 25X/1.05 Plan Apochromat Lambda S, Sil
- 40X/1.15 Apochromat LWD Lambda S, WI
- 60X/1.42 Plan Apochromat Lambda D, Oil
- 100X/1.45 Plan Apochromat Lambda D, Oil
*Available to be put in instrument, please contact Emery to request use. Use the 100X for optimal STED resolution.
Detector units
- DUX with 3 GaAsP and 1 IR-PMT detectors
- NSPARC array of 25 detectors
Software
- NIS Elements confocal package with General Analysis 3 and 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.
![Huygens STED PSF 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.](/content/fralinlifesci_vt_edu/en/Core-Services/advanced-imaging-microscopy/faim-equipment-rates/faim-equipment-nikon-axr-nsparc-stedycon/_jcr_content/content/vtmulticolumn/vt-items_1/adaptiveimage.transform/m-medium/image.png)
![STED dyes for 775 Table of recommended dyes for STED with 775. Abberior dyes for 640 excitation (fixed: STAR RED, STAR 625P, STAR 635; live: LIVE610, SiR) Alternative dyes for 640 excitation (fixed: ATTO647N; live: JF646). Abberior dyes for 561 excitation (fixed: STAR ORANGE; live: LIVE 550, LIVE 560, LIVE 590, LIVE ORANGE, LIVE RED). Alternative dyes for 561 excitation (fixed: AF594, ATTO594; live: ATTO590). Alternative dyes for 488 excitation, long stokes-shift, Abberior dyes (fixed: STAR 460L; live: LIVE 460L). Alternative dyes for 488 excitation, fixed: ATTO490LS.)](/content/fralinlifesci_vt_edu/en/Core-Services/advanced-imaging-microscopy/faim-equipment-rates/faim-equipment-nikon-axr-nsparc-stedycon/_jcr_content/content/adaptiveimage.transform/m-medium/image.png)