New confocal imaging method improves signal-to-noise

DECEMBER 19, 2008--During this week'sAmerican Society for Cell Biology (ASCB) meeting (December 13-17, San Francisco, CA), Nikon Instruments (Melville, NY) launched a new, exclusive confocal imaging method for post acquisition recovery of data normally lost during the course of the experiment. It works by collecting emission photons that are normally rejected by the emission-side pinhole, traditionally used at the 1 airy disk size. The approach, called VAAS Detection, allows for virtual adjusting of the confocality and sensitivity of data post-acquisition, as well as collecting more photons during the initial image acquisition to enable image reconstruction with greatly improved signal-to-noise ratio (SNR).

VAAS, which stands for Virtual Adjustable Aperture System, uses two pinholes to collect emitted photons: a standard-sized one to enable confocality and one with a diameter three-times larger to capture more of the emitted fluorescence without increasing the out of focus background detection. VAAS is an effective method to reduce the out of focus haze signal without reducing the signal from the focal plane.

In using a traditional confocal microscope, images are acquired after adjusting and fixing the diameter of the emission pinhole aperture according to the specimen and instrument conditions. Any emission falling outside of the pinhole is lost. Nikon's VAAS Detection addresses this limitation by collecting photons originating from the selected plane of focus as well as from adjacent planes in a single scan, and independently collecting the signals into independent channels. After the experiment, data can be added back, or the original data can be further deconvolved using this extra collected information.

"The VAAS Detection system leaves no photon behind, satisfying the constant need for clearer images, that are of higher signal-to-noise levels, while enabling the collection of more data than traditional laser point scanning confocal systems." said Stan Schwartz, vice president of Nikon Instruments. "When used with the Ti series of inverted microscopes, the A1 with VAAS Detection offers a complete confocal system that is an ideal platform for a variety of high-level techniques in principal investigator labs and core imaging facilities alike."

The A1 confocal system now offers significant improvements in confocal imaging. In addition to the VAAS detection system, features include a hybrid dual scanner system offering high speed scanning with photoactivation capability and longer cell imaging times. Additionally, the system enables faster and higher sensitivity spectral imaging with better resolution.

VAAS Detection is offered as an upgrade option to Nikon's A1 series of confocal laser point scanning systems, which integrate with the new Ti-E research inverted microscope. The fully-automated confocal imaging system captures high-quality confocal images of cells and molecular events at high speed and with enhanced sensitivity. Suitable for facilities with a broad range of users, the A1 has been designed with advanced optical and electronic technology to provide system flexibility.

More information:
Nikon's A1 confocal series

Posted by Barbara G. Goode, barbarag@pennwell.com.

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