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)
Excitation Modality
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).
Microscopes
Q2 can be interfaced with most commercial epifluorescence and upright microscopes.
Detectors
GaAs detectors, hybrid photomultiplier tubes, avalanche photodiodes.
Specifications for Q2
| Microscope and Coupling |
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| 1p Excitation1 |
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| 2p Excitation1 |
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| Data Acquisition Unit |
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| Detectors |
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| Positioning Controls2 |
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| Software |
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| Computer & Monitor |
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| Power Requirements |
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- 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 Imaging
Confocal 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-FRET
mTurquoise-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 tissue
Laser excitation: 780nm, 80MHz; Emission: 500 - 650 nm; Objective: Olympus 20X / 0.54NA; Image Size:1024x1024 (200 µm x 200 µm)
2p FCS
Fluorescein 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)
- Superresolution
- 3D particle tracking trajectories
- Nanoimaging reconstruction with 20 nm resolution
- Single Molecule Analysis
- Burst Analysis
- FRET and Correlation methods
- PIE-FRET methods
Resources
Configuration
Click on a picture to view it.
Detector Mounts
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Nanoscopic Insights of Amphiphilic Peptide against the Oligomer Assembly Process to Treat Huntington's Disease He, R.‐Y., Lai, X.‐M., Sun, C.‐S., Kung, T.‐S., Hong, J.‐Y., Jheng, Y.‐S., Liao, W.‐N., Chen, J.‐K., Liao, Y.‐F., Tu, P.‐H., Huang, J. J.‐T. Adv. Sci. 2019, 1901165. https://doi.org/10.1002/advs.201901165 |
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Concurrent Exposure of Neutralizing and Non-neutralizing Epitopes on a Single HIV-1 Envelope Structure Ray, K., Mengistu, M., Orlandi, C., Pazgier, M., Lewis, G.K., DeVico, A.L. Front. Immunol., 10:1512, 2019 July. |
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Band-Aligned Polymeric Hole Transport Materials for Extremely Low Energy Loss α-CsPbI3 Perovskite Nanocrystal Solar Cells Yuan, J., Ling, X., Yang, D., Li, F., Zhou, S., Shi, J., Qian, Y., Hu, J., Sun, Y., Yang, Y., Gao, X., Duhm, S., Zhang, Q., Ma, W. Joule, ISSN: 2542-4351, 2018 Sep 10. |
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Composition‐Graded Cesium Lead Halide Perovskite Nanowires with Tunable Dual‐Color Lasing Performance Huang, L., Gao, Q., Sun, L.-D., Dong, H., Shi, S., Cai, T., Liao, Q., Yan, C.-H. Advanced Materials. 2018 May 21. |
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Targeting the Late Stage of HIV-1 Entry for Antibody-Dependent Cellular Cytotoxicity: Structural Basis for Env Epitopes in the C11 Region. Tolbert, W.D., Gohain, N., Alsahafi, N., Van, V., Orlandi, C., Ding, S., Martin, L., Finzi, A., Lewis, G.K., Ray, K., Pazgier, M. Structure. 2017 Nov 7;25(11):1719-1731.e4. doi: 10.1016/j.str.2017.09.009. Epub 2017 Oct 19. |
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Molecular basis for epitope recognition by non-neutralizing anti-gp41 antibody F240. Gohain, N., Tolbert, W.D., Orlandi, C., Richard, J., Ding, S., Chen, X., Bonsor, D.A., Sundberg, E.J., Lu, W., Ray, K., Finzi, A., Lewis, G.K., Pazgier, M. Sci Rep. 2016 Nov 9;6:36685. doi: 10.1038/srep36685. |
