The compact instrument for quantitative cell biology with single-molecule detection
Q2 is a laser scanning nanoscope (LSN) that incorporates several measurement modalities for experimental quantitative biology and material sciences applications requiring the single molecule detection sensitivity.
Key Features of Q2 Laser Scanning Nanoscope:
- Single- and multi-photon excitation on three separate input ports
- Two channel acquisition
- One, computer-controlled-aperture, pinhole
- Fast scanning mirrors
- Powered by VistaVision, a user-friendly software package for the acquisition of confocal images, FLIM/ FRET, RICS, and FFS (FCS, PCH, scanning FCS, N&B)
Three separate input ports allow for simultaneous alignment of one-photon excitation (laser diodes and supercontinuum laser) or multi-photon measurements (fiber laser, Ti:Sapphire laser).
Q2 can be interfaced with most commercial epifluorescence and upright microscopes.
GaAs detectors, hybrid photomultiplier tubes, avalanche photodiodes.
Specifications for Q2
|Microscope and Coupling||
|Data Acquisition Unit||
|Computer & Monitor||
- Q2 was fully evaluated and validated for using ISS laser launcher, Fianium SC-400 laser and Toptica FemtoFiber Pro 2p laser.
- VistaVision provides utilities for measurements in spectroscopy mode (at a single point), raster or orbit scan mode (2D XY), optical sectioning mode (3D), time-lapse mode, stage scan mode for multi wells, or a combination of them.
Measurement Examples for Q2
Confocal ImagingConfocal image of Convallaria (Lily of the valley). Laser excitation wavelength is 561nm (supercontinuum laser). The fluorescence is collected through a bandpass filter centered at 600 nm with 37 nm bandpass. The objective is an Olympus 20X water immersion (NA = 0.54). The pinhole aperture is 2 Airy units. The image size is 1024 x 1024 pixels (200 µm x 200 µm).
Fluorescence Lifetime Imaging (FLIM)The sample is Convallaria (Lily of the valley). The laser excitation wavelength is 488nm from a supercontinuum laser (40 MHz repetition rate, 6 ps pulsewidth). The emission on Ch1 is acquired through a 600/37 nm filter; on Ch2 the emission is acquired through a 525/50nm filter. The objective is Olympus 60X (water immersion; NA = 1.35). The pinhole is 1 Airy unit.
2p FLIM-FRETmTurquoise-5aa-Amber (T5A) expressed in fixed cells (donor-alone control) mTurquoise-5aa-Venus (T5V) expressed in fixed cells (FRET standard) Laser excitation: 780nm, 80MHz; Emission: donor channel: 475/35nm; Objective: Olympus 60X / 1.35NA;
2p FLIM imaging of HE stained pig intestine tissueLaser excitation: 780nm, 80MHz; Emission: 500 - 650 nm; Objective: Olympus 20X / 0.54NA; Image Size:1024x1024 (200 µm x 200 µm)
2p FCSFluorescein in HPLC water PH 7.4.
FFS comprises a whole family of application tools that reveal the inner molecular dynamics upon the detection of fluctuations of molecules due to thermal motion. They include
- FCS, Fluorescence correlation spectroscopy
- FCCS Fluorescence cross-correlation spectroscopy
- PCH, photon counting histogram
Measurements for Q2
- Intensity and Lifetime Imaging (single plane and z-stack)
- 1p or 2p confocal images
- FLIM in frequency-domain or in TCSPC
- Phosphorescence Lifetime Imaging (PLIM)
- Polarization images
- Single Point Measurements
- Intensity, polarization, kinetics, lifetime
- Fluorescence Fluctuations Spectroscopy (FFS)
- Fluorescence Correlation Spectroscopy (FCS)
- Fluorescence Cross-Correlation Spectroscopy (FCCS)
- Photon Counting Histogram (PCH)
- Fluorescence Lifetime Correlation Spectroscopy (FLCS)
- Scanning FCS
- Number & Brightness (N&B)
- RICS (raster imaging correlation spectroscopy)
- 3D particle tracking trajectories
- Nanoimaging reconstruction with 20 nm resolution
- Single Molecule Analysis
- Burst Analysis
- FRET and Correlation methods
- PIE-FRET methods