Ultima 2Pplus

With new advances in field of view, sensitivity, wavelength, and sample accommodation, the Ultima 2Pplus delivers an ideal combination of flexibility, resolution, imaging depth and speed, allowing users to perform simultaneous imaging, stimulation and electrophysiology protocols with greater efficiency and effectivity. The system is designed specifically for intravital imaging, with fully motorized control of the objective X-Y-Z position, as well as two axes of rotation for precise imaging orientation. A second scan path enables simultaneous imaging and photoactivation. The system’s optimized optical train confers exceptional performance to the very edges of the large field.

Class-leading

field of view

Offers a more complete picture with every scan.

High-efficiency

light collection and detection

Images deeper with a greater signal-to-noise ratio.

Simultaneous

imaging and photoactivation

Records cells with exact timing and spatial precision.

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Features

Largest Field of View and Superior Field Uniformity

The Ultima 2Pplus field of view is over 50% larger to facilitate modern optogenetics experiments, which require simultaneous imaging and photostimulation of distant regions in a sample. A new, custom optical design preserves spatial and temporal resolution throughout the entire field, which is essential for researchers to be able to continue their work without changing experimental designs.

Field of view on Ultima 2Pplus is FN 28 mm in comparison to industry standard of FN 18 mm.

High-Sensitivity Deep Imaging

1.3mm deep, 3D-view of pyramidal cells in layer 5. Data collected in one-month-old transgenic Thy1-YFP-H line mouse through a glass-covered cranial window. Data courtesy of Guang Yang, School of Medicine, New York University, New York.

The Ultima 2Pplus light collection path ensures detection of almost all photons scattered from dim, thick and opaque biological samples. The superior collection efficiency is provided with 2-inch or larger optics
throughout the emission path up to the sensitive close-proximity detectors. As two-photon microscopy has become well adopted, emerging three-photon microscopy offers the potential to image even deeper into
samples. Since further infrared wavelengths are scattered even less, three-photon excitation has the promise to be able to image several millimeters into the brain. Ultima 2Pplus already uniquely incorporates this capability to take your research to new levels of potential.

Perfected Photostimulation and Imaging

The Ultima 2Pplus empowers sophisticated optogenetics experiments with Bruker's new Neuralight 3D Ultra Spatial Light Modulator option. Large populations of targets can be stimulated simultaneously in multiple focal planes with computer-generated holograms. During 3D holographic experiments, targets at different depths are imaged with a custom-designed, optically-corrected ETL focusing module that allows for independent Z-positioning of the imaging plane relative to photostimulation.

Read a recent article “Cortico-cortical feedback engages active dendrites in visual cortex” published in Nature from Michael Häusser’s Lab. Bruker's two-photon calcium imaging, eletrotunable lens (ETL), and spatial light modulator (SLM) were used for long-range all-optical connectivity mapping and investigating feedback in the mouse brain.

Optical stimulation of circled cells with Neuralight 3D. 3D hologram of points was created which was then spirally scanned over neuronal cell bodies. Data Courtesy of Adam Packer, Lloyd Russell, Henry Dalgleish, Michael Häusser’s lab, UCL, London.

Applications

Ultima 2Pplus Data Gallery

1.345 mm x 1.345 mm FOV, resonant scanned image of GCaMP expressing cells in mouse brain.

Two-photon imaging of cortical surface with Ultima 2Pplus through a cranial glass window (grey). Acquired with Nikon 16x 0.8 NA objective at 920 nm. Data courtesy of Dustin Herrmann, Mehmet Fisek, Michael Häusser’s lab, UCL, UK.

Co-expression of GCaMP6s (green) and soma-restricted C1V1-mRuby2 (magenta) in layer 5A neurons of mouse visual cortex at a depth of ~400 µm below the pia (right). Both acquired with Nikon 16x 0.8 NA objective at 920 nm. Data courtesy of Dustin Herrmann, Mehmet Fisek, Michael Häusser’s lab, UCL, UK.

Spontaneous fluorescence fluctuations in neurons expressing GCaMP6 in transgenic mouse. Data courtesy of Robert Lees, James Rowland, Adam Packer Lab, University of Oxford, UK.

Large FOV montage (44 on scope with FN28 vs 108 tiles on scope with FN18) of hippocampal slice and details of selected tile. In-house data.

3D-view of volumetric stack recorded using the ETL focusing module. Layer 5B neurons in mouse visual cortex in vivo, labelled with tdTomato. Scale bar 100 µm. Data courtesy of Lisa Bauer, Dustin Herrmann, Mehmet Fisek, Michael Häusser’s lab, UCL, UK.

Co-expression of GCaMP6s and soma-restricted C1V1-mRuby2 in layer 5A neurons of mouse visual cortex. Acquired in vivo, ~400 µm below pia at wavelengths 920 nm and 765 nm. Data courtesy of Dustin Herrmann, Mehmet Fisek, Michael Hausser's lab, UCL, UK

Holographic photostimulation of neurons expressing GCaMP6 and ChRmine in OFC through 4.2 mm grin lens. Data courtesy of Vijay Namboodiri, Charles Zhou, Garret Stuber Lab, University of Washington, WA

Two-photon activation leads to focal constriction. The SLM focused 6 spots at the location indicated by the red arrow. The total stimulation power was ~200 mW spread over the 6 spots with 3 adjacent spots on either side of the vessel. Each spot was scanned in a 12 μm diameter spiral for 100 ms. The white arrow is for comparison. Vessels are labelled with Texas Red Dextran. Speed 4x real time. Data courtesy of Phil O’Herron, University of Augusta, Augusta, GA.

Comparison of FOV between 1x zoom (left) and 0.8x zoom (right) in galvo-galvo mode

Constriction of a penetrating arteriole with the 594 nm LED. 5 pulses, 100 ms duration with 100 ms in between were presented. Speed 4x real time. Data courtesy of Phil O’Herron, University of Augusta, Augusta, GA.

Full-field activation with the 594 nm LED in a PDGFRβ/ReaChR mouse. Image is looking down at pial surface vessels in the neocortex labelled with FITC dextran. A single 100 ms pulse from the 549 nm LED was used to stimulate the vessels. The dark flash indicates the time of the optical stimulation. The image darkens because the detectors are briefly blocked by mechanical shutters to protect them from the intense stimulation light. Speed 4x real time. Data courtesy of Phil O’Herron, University of Augusta, Augusta, GA.

EGFP-expressing dendritic epidermal T cells (green) in the skin of a mouse. Intravenously injected Rhodamin 6G demarcates the dermal-epidermal border (red). Hair shafts (purple) are rooted in hair follicles revealed by their sebaceous glands (blue autofluorescence). Data courtesy of Dr. Thorsten Mempel, MGH Harvard.

mCherry-expressing dendritic cells (red) in the female genital tract of a CD11c-mCherry transgenic reporter mouse. The animal was intravenously injected with Hoechst 33342 to stain all nucleated cells (blue). The epithelial cells visualized with autofluorescence signal (green/turquois hue). Data courtesy of Dr. Thorsten Mempel, MGH Harvard.

EGFP-expressing dendritic epidermal T cells (green) in the skin of a mouse. Intravenously injected Rhodamin 6G demarcates the dermal-epidermal border (red). Hair shafts (purple) are rooted in hair follicles revealed by their sebaceous glands (blue autofluorescence). Data courtesy of Dr. Thorsten Mempel, MGH Harvard.

GFP-expressing T cells (green) accumulating around a pancreatic islet in a mouse developing autoimmune pancreatitis ultimately leading to diabetes. Data courtesy of Dr. Thorsten Mempel, MGH Harvard.

GFP-expressing cytotoxic T lymphocytes (green) and tdTomato-expressing regulatory T cells (red) infiltrating a mouse colon carcinoma implanted into a dorsal skinfold chamber. Tumor cell nuclei (blue) are tagged through expression of a Cerulean-histone H2B fusion protein. Blood vessels (white) are highlighted through intravenous injection of QTracker 655 quantum dots. Data courtesy of Dr. Thorsten Mempel, MGH Harvard.

Specifications

SCAN HEAD
Scanning Method	Matched pair of 6 mm Cambridge galvanometers with raster and spiral scanning capabilities
Field of View	~1.375 mm x 1.375 mm with 16x objective (≤28 mm FN)
Scan Speed	Raster scan: 1.65 fps at 512 x 512, >12 fps at 64 Spiral scan: 6 fps at 512 x 512, ~30 fps at 64 x 64
Scan Customization	User-definable straight, freehand and circular (infinite) linescan with included software; User-definable pixels/line and lines/scan from 1 to 2048; ≤120x scan zoom; 360° of scan rotation; Point scan
Uncaging Option	Second set of matched 3 mm or 6 mm Cambridge galvanometers in same scan head provide high-precision visible or multiphoton laser sample photomanipulation
High-Speed Imaging Option	8 kHz resonant galvanometer; ~30 fps at 512 x 512, >1300 fps at 512 x 8 region of interest
DETECTORS
Reflected Non-Descanned	1 to 4 hand-picked Hamamatsu Multi-Alkali PMTs; Upgradeable to high-sensitivity Hamamatsu GaAsP PMTs
Transmitted Non-Descanned	1 or 2 hand-picked Hamamatsu Multi-Alkali PMTs; Upgradeable to high-sensitivity Hamamatsu GaAsP PMTs
Dodt	Single Hamamatsu PMT for DIC-like image collection
Transmitted	Single Hamamatsu PMT for transmitted light image collection
Camera	Standard C-mount camera port built into scan head
OPTICAL INPUTS
Multiphoton Laser	Optimized for multiphoton laser input from 690 to 1700 nm
Epifluorescence	High-Powered LED in epifluorescence turret
Visible Laser	Fibered laser inputs for visible laser introduction
Ultima Laser Rating	Class 1 (Contains Class 3b and Class 4 lasers) with light box
Helios Laser Rating	Class 3b
LED	Full-field photoactivation with LED module
MOTOR CONTROL
Bruker X, Y Stage	Variable height with ~15 cm X and ~7.5 cm Y movement and ~0.3 μm step size
Bruker X, Y Microscope Base	Fine and coarse-movement platform for scope with ~35 mm travel and 0.1 μm step size
Bruker Z-Focus	Range of 30 mm with ~0.2 μm step size
Bruker Z-Piezo	Travel range ≤1000 μm with 0.05 μm step size
Bruker Orbital Nosepiece	Motorized control of objective angle and rotation
SOFTWARE
Prairie View Imaging	Turnkey intuitive and customizable operation for imaging
Z-Series	Easy creation of depth stacks with user-customizable slice number, step size and laser power
T-Series	Easy creation of complex series involving Z-Series and triggered images
Stage Montage	Atlas Imaging simplifies setup and optimizes acquisition of 2D and 3D stage montages
Peripherals Integration	Wavelength and power control available for multiphoton and visible laser launches
Photoactivation	User-defined points and regions with synchronized laser modulation
Regions of Interest	User-defined regions for faster scanning capabilities
Brightness Over Time	Intensity mapping and plotting for user-defined regions over time
Voltage Inputs/Outputs	Signal inputs and outputs for electrophysiological experiments, stimulus control, and synchronization with external devices

Webinars

Recent Multiphoton Microscopy Webinars

Testimonials

Hear What Our Customers Have to Say

We have the 2Pplus - the scope is well-designed and the software easy to use. Their scopes are very flexible in design if you wanted to add additional optics, components, or software functionality. Bruker shines with respect to customer service… they have several great service experts who usually respond within a day or two with insightful solutions. Further, the support is highly personalized; not before long after purchasing the microscope, we were paired with specialists who intimately familiarized themselves with our applications.

Charles Zhou, Ph.D., UW Center in Neurobiology of Addiction, Pain, and Emotion

We appreciate the modular nature of the Ultima hardware and software. The systems now available range from basic imaging systems to the dual-laser photo-stimulation multi-mode imaging workstation. We have continued updating many of our 14 machines, including our ten year old systems. The Team Viewer connection and support is a huge asset for novice owners and for significant software changes.

David Wokosin, Ph.D., Northwestern University

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