Confocal Microscopy In-Sync with Nature

Even the most rapid biological processes can now be captured at ultra high resolution as a result of the launch of a new range of confocal laser scanner systems by Nikon Instruments. Two models are available; the fully automated A1 and the high specification A1R. The A1 utilises conventional paired galvonometers producing high resolution images (up to 4096 x 4096 pixels), whilst the A1R incorporates a unique hybrid scanner system (offering frame rates of 30fps, 512 x 512 pixels). This facilitates ultra-high-speed imaging with unsurpassed image quality. Furthermore, the hybrid scanner enables simultaneous photo-activation and imaging, critical for unveiling cell dynamics and interactions.

The new systems are a natural complement to the recently launched Ti inverted microscope, particularly when coupled with Nikon’s patented Perfect Focus System, essential for eliminating focal drift. Together they set a new standard for advanced time-lapse studies of rapid cellular interactions to literally bring biological imaging to life.

The high specification A1R features an innovative hybrid resonant-non-resonant scanning system creating a powerful new tool for live cell imaging. The scanners are used in tandem; the resonant scanner capable of capturing images at 30fps (512 x 512 pixels), is employed to capture images whilst the non-resonant scanner photoactivates or photobleaches the specimen. Additional functionality of the hybrid scanning system enables images to be captured at speeds of up to 420fps (512 x 32 pixels). The A1R also boasts an optical pixel clock generator which is used to produce ultra stable clock sync pulses, offering images that are completely even in intensity without distortion even at high speed.

Higher signal to noise imaging is achieved with both models thanks to a 30% increase in signal capture (compared to standard systems). This is due to the inclusion of the industry’s first low-incidence-angle dichroic mirror giving rise to an increase in fluorescence efficiency. Furthermore, the A1R and the A1 have been designed to incorporate innovative variable hexagonal shaped pinholes that considerably sharpen image quality. By not clipping the airy disc (as with square pinholes), more light is passed through the pinhole improving optical efficiency. Coupled with the variable Z resolution, brightness, noise and scanning times can all be optimised. Image quality is further enhanced by hyper double sampling, whereby PMT (photomultiplier tube) noise is sampled twice and eliminated, reducing noise on images by a third.

An additional benefit of the series is its ability to be used in conjunction with Controlled Light Exposure Microscopy (CLEM). CLEM is a system for automatically monitoring and varying laser illumination during time-lapse studies in order to minimise the risk of cell degradation, death or bleaching.

As with all Nikon’s microscopes, the series is empowered with the company’s NIS- Elements software for image capture, processing and analysis. This means that procedures such as, confocal, TIRF and widefield imaging, can all be performed on the same microscope with the same software. Moreover the system is provided, as standard, with the analysis software for FRAP and FRET.