Overview
The Bruker Opterra Multipoint Scanning Confocal Microscope is engineered for live-cell fluorescence microscopy, combining high resolution typical of confocal systems with the speed of wide-field imaging. It uses a CCD camera detector and offers selectable aperture sizes to optimize speed and resolution for diverse experimental needs. Its unique optical design enables spectral imaging of dynamic processes with minimal photobleaching and phototoxicity, making it ideal for advanced live-cell studies such as protein localization, intracellular ion imaging, and cellular dynamics.

Key Features and Performance
- High-speed, multidimensional imaging of live cells and small organisms.
- Photoactivation module tailored for cellular kinetic studies.
- Intuitive, powerful Prairie View software for complex imaging protocols.
- Motorized aperture plate with 7 selectable apertures (3 pinholes and 4 slits) allowing optimization between resolution and speed without hardware changes.
- One-dimensional pinhole arrays reduce crosstalk compared to two-dimensional spinning disk arrays, resulting in sharper images with better resolution and imaging depth.
- Spectral imaging at 4 frames per second with 15 spectral channels per frame at 512x512 pixel resolution.
- Simultaneous photoactivation and imaging without emission signal loss.

Comparison with Spinning Disk Systems
The Opterra system offers advantages over traditional spinning disk confocals, including:
- Higher speed operation with slit apertures (up to 1000 fps).
- Reduced photobleaching and phototoxicity.
- Improved axial resolution and imaging depth due to linear pinhole configuration.
- Software-controlled aperture selection without hardware modifications.
- Enhanced spectral imaging capabilities.
- Integrated photoactivation with no emission loss, supporting advanced photomanipulation techniques.

Software and Data Acquisition
Powered by Prairie View software, the system supports:
- User-friendly setup of time lapse, Z-series, stage montages, and multidimensional acquisitions.
- Integration of external devices via triggers and analog signals.
- Atlas Imaging module for simplified multifield and stage montage imaging with intuitive navigation in X, Y, and Z.
- Real-time spectral data processing and visualization.
- Control of photostimulation protocols including points, lines, rectangles, arbitrary shapes, and spiral activation.
- Support for complex photoactivation experiments such as FRAP, photoconversion, optogenetics, and uncaging.

Applications
Typical applications include:
- Protein localization and trafficking
- Vesicle trafficking
- Mitosis and microtubule dynamics
- Organelle trafficking
- Cell migration
- Intracellular ion concentration measurements
- FRAP and photoconversion
- Cell membrane wound healing
- Photoactivation and optogenetics

Optical and Mechanical Specifications
- Scanning method: Combination of galvanometer and piezoelectric crystal scanning.
- Apertures: Motorized plate with 3 pinhole sizes (30, 45, 60 µm) and 4 slit widths (22, 35, 50, 70 µm).
- Scan speeds: Up to 50 fps in pinhole mode; up to 1000 fps in slit mode with appropriate camera.
- Filters: Motorized 6-position emission filter wheel; custom dichroics and polychroics.
- Triggering: Frame trigger in/out and 4 auxiliary device triggers.
- Detector: CCD camera with support for Photometrics and Q-Imaging cameras; standard Evolve Delta EMCCD or Rolera EM-C2 EMCCD.
- Optical inputs: Helios launch with up to 5 diode laser lines, analog modulated with digital blanking; fiber optic input.
- Microscope platforms: Nikon TI-E inverted and Nikon FN-1 upright with software control of turret and Z-motor.
- Specimen stage: ASI XY stage with piezo focus insert (300 µm range) for TI-E; Bruker Slim Stage for FN-1.
- Z focus: Internal focus motor and piezo for TI-E; Bruker Z-focus motor and piezo-driven objective focus for NI-E.
- Photoactivation scanner: Single-pair galvanometer (3 mm) for high-speed photoactivation, photobleaching, and ablation.
- Analog I/O: Up to 8 analog outputs and 8 analog inputs for device control and signal recording.
- Software: Prairie View Imaging Software on Windows 7 64-bit.
- Computer: Intel Quad Core processor, multiple SSDs for OS, software, and data, 32GB RAM.

Summary
The Bruker Opterra Multipoint Scanning Confocal Microscope is a versatile, high-performance imaging platform optimized for live-cell fluorescence microscopy. Its unique optical design, flexible aperture selection, and integrated photoactivation capabilities provide superior imaging speed, resolution, and spectral imaging compared to traditional spinning disk systems. Coupled with powerful and user-friendly software, it supports a wide range of advanced biological applications requiring dynamic, multidimensional imaging with minimal photodamage.