Q2Modular Confocal Microscopefor FLIM and FFS

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Overview of Q2

A compact and adaptive microscope system for comprehensive time-resolved confocal imaging and FFS measurements, the Q2 can be equipped with single-photon and multi-photon excitation and up to five detection channels to meet the single molecule sensitivity challenges and the fast data acquisition required for live cell imaging and material science applications.

The detectors are conveniently mounted on sturdy baseplates holders that are coupled to the Q2. Cubes holding dichroic filters are located at the channels junctions to separate the wavelength ranges directed to each detector; filters in front of each detector select the spectral wavelength region. The cube accommodates polarizing beam splitters for polarization acquisition.

Holder units for 2, 3 and 4 detectors are available. Detectors type may be different on a holder; for instance, a 4-detector unit may carry two SPADs and two hybrid PMTs. A fiber port can replace a detector coupling hardware to connect a spectrograph.

Key Features of Q2

Modularity

Modularity

A selection of laser wavelengths, detectors, number of detection channels, & microscopes.

Superb Confocal Images

Superb Confocal Images

Lifetime Measurements

Lifetime Measurements

From 100 ps to 100 ms

Autofluorescence Signal Unmixing

Unmixing the Signal from Autofluorescence Using the Phasor Plot

Single Molecule Studies

Single Molecule Studies

Detection of Full-Scale FCS

Affordable Solution

Affordable Solution

Superb Confocal Images

Stained Bovine Pulmonary Artery Endothelial (BPAE) cells

Bovine Pulmonary Artery Endothelial (BPAE) cells stained with Mito-Tracker Red CMXRos and Alexa Fluor 488 Phalloidin

Intensity and Lifetime Imaging

Convallaria (lily of the valley) fluorescence intensity image Convallaria (lily of the valley) fluorescence lifetime image

Convallaria (lily of the Valley); Intensity image (first) and lifetime image (last). Image size is 1024 x 1024 pixels (200 µm x 200 µm). Microscope is IX-73 by Olympus using a 60X, NA=1.35 objective. Excitation is 488 nm, emission is collected on CH1 through a 600/37 nm filter, and CH2 through a 525/50 nm filter.

Unmixing the Signal from Autofluorescence in Live Cells Using the Phasor Plot

Raw image, umixed image, and phasor plot of ACD with NADH

Acridonylalanine (ACD) is a fluorescent amino acid and its spectra significantly overlap with the endogenous autofluorescent molecule NADH, making imaging of ACD in live cells be likely contaminated by cellular NADH signals. As the lifetime of ACD (about 15 ns) is very different than the lifetime of NADH (< 5 ns), the time-resolved (TR) unmixing procedure is instrumental in producing two separate images for ACD and NADH, respectively.

Single-molecule studies: Detection of Full-Scale FCS

Continuous wave for Rhodamine 110 in water with excitation at 488 nm

The full time-scale FCS allows for investigation of single molecules photophysical dynamics from picoseconds to seconds. The autocorrelation function determined by FCS provides the translational diffusion of molecules and the chemical or photophysical effects on the molecules, which typically occur in the time scale longer than 1 µs. The extension of the autocorrelation function to the picosecond time scale range provides additional information about the single molecule or nanoparticle: the rotational diffusion times; antibunching. Data in the figure were collected on Rhodamine 110 in water with a continuous wave excitation at 488 nm.

Product Specifications for Q2

Microscope and Coupling
  • Evident (Olympus), Nikon, Zeiss and Leica
  • Inverted and upright
  • Left side port and back port
1p Excitation*
  • ISS laser launcher (models for 3-, 4-, and 6-lasers), wavelengths available from 375 nm to 1,000 nm; pulse interleave excitation (PIE)
  • Supercontinuum lasers, wavelength from 400 nm to 950 nm
2p Excitation*
  • Ultra fast femtosecond pulse Ti:Sapphire lasers
  • Ultra fast femtosecond pulse fiber lasers
Galvanometer scanner
  • 2 silver-coated galvanometer scanning mirrors
  • Clear optical surface: 3 mm or 5 mm
  • Maximum scan rate: 5 KHz for 3 mm and 1 KHz for 5 mm
  • Scanning resolution: 64 x 64 to 4096 x 4096 pixels
  • Scanning mode: Pt, Xt, XZ, XY, XZt, XYt, XYZ
  • ROI scanning: rectangle, ellipse, polygon, line
Positioning Controls**
  • ISS 3-axis control unit
  • ISS XY galvo scanning mirrors control unit
  • ISS Z-piezo control unit
  • Microscope built-in focusing control module
  • Automatic XY stages (ASI, Prior)
  • XYZ piezo stages (MadCity, PI)
Pinhole
  • Single, variable-aperture pinhole; diameter from 20 µm to 1,000 µm
Detectors
  • Cooled GaAsP and GaAs PMT
  • Cooled Hybrid PMTs
  • SPADs
Data Acquisition Unit
  • FastFLIM (Digital Frequency Domain FLIM)
  • SWISS TCSPC card (Time Domain FLIM)
Software
  • VistaVision
Computer & Monitor
  • High-performance Processor, 32 GB RAM
  • 32" monitor, 2556 x 1440 resolution
Operating System
  • Windows 10, 64-bit
Power Requirements
  • Universal power input: 110 - 240 V, 50/60 Hz, 400 VAC
Dimensions (mm)
  • 420 (L) x 330 (W) x 150 (H)
Weight (kg)
  • 13.5 (without detectors)
Notes
  • *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.

Measurements for Q2

Confocal Intensity and Lifetime Imaging
Steady-state and Time-resolved Polarization Anisotropy Imaging
Fluorescence Fluctuations Spectroscopy (FFS)
Particle tracking (optional)
Single Molecule FRET Bursts Analysis
Antibunching
Download Data Sheet

Configuration for Q2

Configuration of Q2 Modular Laser Scanning Microscope

Product Accessories for Q2

Additional Product Options

  • Perfusion system stabilizes conditions
    Perfusion System

    A peristaltic pump supplies the stage with a solution for keeping the sample conditions (temperature, pH, etc...) stable.

  • Irrigation system prevents drying up samples
    Irrigation System

    When using water objectives for prolonged measurements, it prevents the liquid drying up.

  • Maintain focus automatically
    Auto Focus Maintaining

    It keeps the focus position of the objective for hours using an active feedback to counter drifts.

  • Maintain environmental conditions for cell cultures
    Sample temperature control

    Stage top incubator or a full enclosure to maintain the environmental conditions of cell cultures.

  • Epifluorescence lamp
    Epifluorescence Lamp

    Visualize your sample with the Epi module. Select as light source either an arc lamp or an LED and the suitable filter cubes to add to the microscope cassette.

  • Connect to an atomic force microscope
    Atomic Force Microscope (AFM)

    Fully integrated for the models:
    NanoWizard by JPK-Bruker
    Resolve by Bruker
    For other models contact ISS.

Product Software for Q2

Screenshot of VistaVision working on the Q2 Modular Microscope

VistaVision

VistaVision is a complete software package for confocal microscopy applications including instrument control, data acquisition and data processing. Easy to use, the software has been developed in modular components that can be activated when a specific instrument configuration is selected. The modules include:

  • FLIM/PLIM Imaging
  • FFS
  • smFRET
  • Particle Tracking
Learn More

Product Resources