Alba STEDFLIM/FFS Laser Scanning MicroscopeSuper Resolution at Your Fingertips!

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Overview of Alba STED

Stimulated Emission Depletion (STED) is a powerful super resolution microscopy technique that allows for the observation of fluorescence structures with spatial resolution below the diffraction limit. The Alba STED uses the pulsed excitation and pulsed emission approach (pSTED) to achieve lateral resolution of 60 - 100 nm. An additional increase on the resolution is provided by the lifetime information acquired through FastFLIM (the digital frequency domain lifetime imaging) and using the phasor plot: the photons emitted by the fluorophores activated by the excitation laser only (inherent fluorescence decay) are efficiently separated from the photons emitted by the fluorophores partially or fully activated by the STED laser (featuring shorter decay times). A resolution of 30 nm is measured on standard samples. Moreover, the combination of pSTED and lifetime analysis through the phasor plot allows to separate two fluorophores featuring similar emission patterns but having different decay times (dual label excitation).

Key Features of Alba STED

High dynamic range of up 60 million counts/s

High Dynamic Range

Speed and precision in FastFLIM

Fast Image Acquisition

(Dwell Time: 0.2 µs)

Pulsed excitation and pulsed STED in FastFLIM

pSTED

(Pulsed Excitation and Pulsed STED)
w/ FastFLIM for Time-Resolved Acquisition

Improved image resolution using lifetime information and the phasor plot

Improved Image Resolution
Using Lifetime Information
and the Phasor Plot

Dual label excitation on a phasor plot

Dual Label Excitation

The Data is Clear!

Confocal image of actin in neuronsConfocal
pSTED image of actin in neuronspSTED
Actin labeled by SiR in Neuron cells
(courtesy of Dr. Paul Selvin, University of Illinois at Urbana-Champaign)
Confocal image of mitochondria in HeLa cellsConfocal
pSTED image of mitochondria in HeLa cellspSTED
Mitochondria in HeLa cells labeled by Atto647N
(courtesy of Dr. Paul Selvin, University of Illinois at Urbana-Champaign)
Confocal image of microtubule in HeLa cellConfocal
pSTED image of microtubule in HeLa cellpSTED
Live HeLa Cell microtubule labeled by SiR
(courtesy of Dr. Paul Selvin, University of Illinois at Urbana-Champaign)
Confocal image of HSV1Confocal
pSTED image of HSV1pSTED
HSV1 GFP Atto647N
(courtesy of Dr. Alex Evilevitch, University of Illinois at Urbana-Champaign)
Confocal image of nucleus pores in COS-7 cellsConfocal
pSTED image of nucleus pores in COS-7 cellspSTED
Nucleus pores from COS-7 cells stained with anti-NUP STAR635P dye

Product Specifications for Alba STED

Instrument Features
  • Individual pinholes on each acquisition channel
  • Computer-controlled selection of the pinhole variable aperture
  • Computer-controlled positioning of the pinhole in the imaging plane
  • Single-photon or multi-photon excitation
  • Up to four channel data acquisition
  • Auxiliary port for camera
Microscope and Coupling
  • Olympus, Nikon, Zeiss
  • Inverted and upright
  • Left side port and back port
STED Laser 1
  • Pulsed, 775 ± 2 nm
  • Average output power: 1.2 W at 80 MHz
  • Pulsewidth: 700 ± 200 ps
  • Repetition rate: 20 - 80 MHz
  • Beam quality (TEM00): M2 < 1.3
Or STED Laser 2
  • Pulsed, 592 ± 2 nm
  • Average output power: 1.2 W at 80 MHz
  • Pulsewidth: 700 ± 200 ps
  • Repetition rate: 20 - 80 MHz
  • Beam quality (TEM00): M2 < 1.3
Excitation Laser
  • Pulsed, 640 nm
  • Pulsewidth (at medium power): 40 - 90 ps
  • Repetition rate: from 500 KHz to 100 MHz
  • Power (at 40 MHz): up to 4 mW
Additional Lasers
  • Single photon lasers housed in a laser launcher with computer-control of laser intensity and shutter
Laser Launcher
  • Models for 3-, 4-, 6-laser. Light is delivered to the microscope through a single-mode fiber optic.
Galvanometer Scanner
  • Two silver-coated galvanometer scanning mirrors
  • Clear optical surface: 5 mm
  • Maximum scan rate: 1 KHz
  • 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 1000 µm
Detectors
  • Cooled GaAsP and GaAs PMT
  • Cooled Hybrid PMTs
  • SPADs
Dichroic Filters
  • For single-photon excitation: 1-, 2-, 3-band filters
  • For multi-photon excitation
Polarizer
  • Cube beam splitter, wavelength range: 450 - 1100 nm; extinction ratio: 10,000:1 at ±3 degrees
Data Acquisition Unit
  • FastFLIM (Digital Frequency Domain FLIM)
Software
  • VistaVision
Computer & Monitor
  • High-performance Processor, 32 GB RAM, Windows 10, 64-bit
  • 32" monitor, 2556 x 1440 resolution
Power Requirements
  • Universal power input: 110 - 240 V, 50/60 Hz, 400 VAC
Dimensions
  • 885 mm (L) x 600 mm (W) x 330 mm (H)
Weight
  • 40 kg (without microscope)

Measurements for Alba STED

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

Configuration for Alba STED

Alba STED Schematic Diagram

Product Accessories for Alba STED

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 Alba STED

Screenshot showcasing VistaVision

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