CHOOSE FROM THREE CONFIGURATIONS
Available in upright, inverted, and open (movable objective) configurations, VIVO Multiphoton
is highly flexible and can be customized to specific imaging needs.
UPRIGHT
Slice electrophysiology, zebrafish, Drosophila
INVERTED
Coverslip-mounted specimens, full environmental control
OPEN
In vivo, virtual reality
Flexibility
mBEAM™
Software-controlled beam expander adjusts beam diameter to fill variable objective back aperture sizes.
TTL SYNCHRONIZATION
Integration of electrophysiology and virtual reality hardware.
FAST-GATED PMTs
Sub-millisecond switching between imaging and photostimulation.
SIDEPORT™
Adds 2 telecentric inputs for photostimulation and ablation on the upright configuration.
Scanning + Photostimulation
Vector™ RS+ hybrid scanner combines the speed of resonant scanning with the flexibility of dual galvanometers.
Rapid switching between 30 fps full-frame resonant scanning and spiral or spot photostimulation.
Pulse-triggered digitization ensures maximum signal collection by syncing acquisition to laser output.
Core components
Computer generated holography
with Phasor
Optogenetic photostimulation and uncaging with the highest available temporal resolution and simultaneous illumination of non-adjacent regions.
Spatial light modulator (SLM) creates a holographic illumination pattern via shapes, hand-drawn regions, and diffraction-limited spots.
Capable of simultaneous stimulation in multiple 3D locations above and below the plane of focus.
Choose from a combination of up to 7 visible wavelengths or a high power fixed-wavelength femtosecond laser for near-IR stimulation and uncaging.
Optional Temporal Focusing for Phasor 2-Photon confines the axial thickness of large illumination regions to < 20 µm.
Rapid 4D imaging
Dynamic, continuous beam diameter and laser power adjustments for optimal signal collection.
Piezo objective collar eliminates delay between frames with continuous, synchronized Z movement.
SlideBook algorithms track the sample in X, Y and Z to maintain focus and alignment.
2pFLIM
2P Time Correlated Single Photon Counting (TCSPC)
Fluorescence Lifetime Imaging Microscopy (FLIM)
SlideBook module. Utilizes standard GaAsP detectors or optional hybrid PMT / APD detectors.
PM-ER interaction in dendritic spines imaged with 2pFLIM
MBL Neurobiology, 2015
Customize your system
CORE COMPONENTS
Microscope body
and objective
Modified Zeiss Axio Examiner Z1
frame body and 20x / 1.0
long working distance water
immersion objective.
2P Vector Scanner
High-speed 6 fps @ 512 x 512 and
3,000 lps with bi-directional scanning
for maximum acquisition speed.
Patterns include frame, line, curve,
multi-region, multi-point, and precisely
timed excursions for photostimulation
and laser ablation or scission.
Tunable TI:
Sapphire Lasers
Pockels cell control of up to two
lasers with 200 kHz response.
SlideBook 6
Digital microscopy software for
advanced acquisition and analysis
software for biomedical research
that is easy-to-use and powerful.
Piezo Z Focus
400 micron total travel with 1.25 nm
step size and 5 nm repeatability
(microscope focus travel 13 mm with
50 nm step size).
mBeam
Beam expansion and modification
for shaping multiphoton laser sources
and coupling efficiently to various
objective back aperture diameters.
DETECTORS
Nosepiece
Dual 14-bit GaAsP PMTs collect
reflected light close to the
sample for maximal sensitivity.
IR transmission option
Bialkali PMT in unit mounted
at transmitted illumination port
(shared with transmitted LED
by a beamsplitter).
Remote PMT array
High sensitivity Peltier-cooled
GaAsP PMTs allow for
additional acquisition channels.
Sub-staged detectors
Up to 2 GaAsP PMTs located
below the stage to collect maximal
transmitted light via 1.2NA water
immersion condenser.
STAGES
Platform stage
Stable, fixed, large working area stage compatible
with mounting multiple manipulators and
large trays for tissue and animal presentation.
Luigs & Neumann
Highly flexible stages, specimen platforms
and manipulators for complex specimen
micromanipulation.
ADVANCED TECHNOLOGIES
Resonant scanning
High speed resonant scanning
of up to 30 fps at 2048 x 2048
in frame and line patterns.
1P and 2P Phasor
Digital holography pattern
photomanipulation in X,Y and Z
without intensity loss.
2pFLIM
Using either integrated GaAsP
detectors or hybrid PMT/APDs, photon
counting hardware, and the SlideBook™
2pFLIM module.
Temporal focusing
for 2P Phasor
Disperses laser illumination above and below
the plane of focus dramatically reducing
unwanted fluorescence excitation and ensuring
Phasor illumination of larger regions or cells deep
in scattering tissue are still axially confined to
the desired target.
Widefield imaging
Camera options include EMCCDs and
high-resolutions CMOS cameras for
fast switching between widefield and
multiphoton imaging.
Specifications
BASE OPTIONS
Upright, inverted, or open (movable objective)
SCANNER
Vector RS+ hybrid scanner:
• Merges the speed of resonant scanning with the flexibility of dual galvanometers
• Fast scanning can be paused for sub-millisecond switching to spiral or spot photostimulation
Vector dual galvanometer scanner:
• Up to 4 frames per second with bidirectional scanning at 512 x 512, maximum frame size 2048 x 2048
• Up to 1500 lines per second (unidirectional)
• Frame, line, and curve scanning patterns
• Point or spiral photostimulation
Vector RS resonant scanner:
• Up to 30 frames per second at 512 x 512, maximum frame size 2048 x 2048
DETECTORS
Two GaAsP PMTs mounted in custom nosepiece for close proximity to the back aperture
One or 2 substage GaAsP detectors for additional signal collection
Up to 4 multialkali or GaAsP (with optional cooling) detectors in a custom c-mount array
Red-shifted multialkali PMT for transmitted IR imaging
PHOTOSTIMULATION
Computer-generated holography:
• Phasor or Phasor 2-Photon
• Simultaneous illumination of multiple arbitrary regions
• 2-Photon uses high-powered fixed-wavelength NIR laser or fixed line of dual-line NIR laser
• Temporal Focusing for Phasor 2-Photon confines z illumination to 20µm for holograms with diameter 20-100µm
Scanning:
• Vector for point or spiral stimulation
OPTICS
Custom-designed SidePort for combining multiple inputs
Automated beam expansion (0.25x-3.9x) to accommodate multiple objectives
Compatible with a variety of high-NA, long-working distance (up to 2.4mm) objectives
Available DIC, Dodt contrast or phase contrast
STAGE
Fully-integrated, software-controlled XY stageRange of stages with available motorization, linear-encoding, interchangeable bridges, and integrated micromanipulators
LASER
Full integration and software control of tunable NIR lasers including Coherent and SpectraPhysicsPower modulation and blanking controlled by electro-optic modulator
Publication Spotlight
GCaMP imaging of visual responses in adult Drosophila. Frye lab, UCLA
Keleş and Frye, 2017. “Object-Detecting Neurons in Drosophila.” Curr Biol. 2017 Jan 30. pii: S0960-9822(17)30012-X.
https://www.ncbi.nlm.nih.gov/pubmed/28190726
Imaging of electrical activity in an isolated guinea pig heart preparation. Department of Circulation and Medical Imaging, Norwegian University of Science and Technology
Weinberger et al., 2016. “Cardiac repair in guinea pigs with human engineered heart tissue from induced pluripotent stem cells.” Sci Transl Med. 2016 Nov 2;8(363):363ra148.
https://www.ncbi.nlm.nih.gov/pubmed/27807283
2-photon stimulation of Chrimson in mouse cortical slices using Phasor 2-Photon, imaging with VIVO Multiphoton on a Sutter Movable Objective Microscope. Adesnik Lab, UC Berkeley
Merel et al., 2016. “Bayseian method for event analysis of intracellular currents.” J Neurosci Methods. 2016 Aug 30;269:21-32.
https://www.ncbi.nlm.nih.gov/pubmed/27208694
In vivo imaging in mouse brain through an implanted GRIN lens. Murray Lab, Louisiana Tech University
Voziyanov et al., 2016. “TRIO Platform: A Novel Low Profile In vivo Imaging Support and Restraint System for Mice.” Front Neurosci. 2016 Apr 25;10:169.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4842766/
GCaMP imaging in spinal motor neurons of awake, behaving zebrafish. Wyart lab, Institut du Cerveau et de la Moelle Epinière (ICM)
Böhm et al., 2016. “CSF-contacting neurons regulate locomotion by relaying mechanical stimuli to spinal circuits.” Nat. Commun. 2015 Mar 7;7:10866.
http://www.nature.com/articles/ncomms10866
Calcium imaging in mouse brainstem slices in conjunction with electrophysiology and GABA uncaging using 488 laser line. Kandler lab, University of Pittsburgh
Weisz et al., 2016. “Excitation by Axon Terminal GABA Spillover in a Sound Localization Circuit.” The Journal of Neuroscience, 20 January 2016, 36(3):911-925.
Imaging of cleared mouse spinal cord tissue. Steward Lab, UC Irvine
Willenberg and Steward, 2015. “Nonspecific labeling limits the utility of Cre-Lox bred CST-YFP mice for studies of corticospinal tract regeneration.” J Comp Neurol. 2015 Dec 15;523(18):2665-82. doi: 10.1002/cne.23809.
https://www.ncbi.nlm.nih.gov/pubmed/25976033
GCaMP imaging of visual responses in adult Drosophila. Frye Lab, UCLA
Aptekar et al., 2015. “Neurons forming optic glomeruli compute figure-ground discriminations in Drosophila.” The Journal of Neuroscience, 13 May 2015, 35(19): 7587-7599.
http://www.jneurosci.org/content/35/19/7587.long
GCaMP imaging in the optic lobe of adult Drosophila during visual stimulation. Frye lab, UCLA
Wasserman et al., 2015. “Olfactory Neuromodulation of Motion Vision Circuitry in Drosophila.” Current Biology , Volume 25 , Issue 4 , 467 – 472.
http://dx.doi.org/10.1016/j.cub.2014.12.012
Morphological imaging of labeled neurons in the auditory pathway in mouse brainstem slices. Kandler lab, University of Pittsburgh
Clause et al., 2014. “The precise temporal pattern of prehearing spontaneous activity is necessary for tonotopic map refinement.” Neuron , Volume 82 , Issue 4 , 822 – 835.
http://dx.doi.org/10.1016/j.neuron.2014.04.001
2P morphological imaging in rat brain slice combined with 1P ChR excitation and electrophysiology. Otis lab, UCLA
Mathews et al., 2012. “Effects of climbing fiber driven inhibition on Purkinje neuron spiking.” The Journal of Neuroscience, 12 December 2012, 32(50): 17988-17997.