Gentle Imaging at the Speed of Life with Lattice Illumination

Sven Terclavers

Carl Zeiss Microscopy, LLC

Renee Dalrymple, Ph.D.

Carl Zeiss Microscopy, LLC

Abstract

In order to best understand the world around us it is necessary to observe microscopic specimins in as natural a state as possible. This requires a transition from imaging fixed to live specimins and expanding from 2D to 3D model organisms. The drive towards live-cell imaging over long timeframes and at high volume speeds brings new challenges. There is evidence that traditional imaging techniques can influence the behavior of specimins due to phototoxicity, thus affecting the integrity of results. The most influential technological breakthroughs which address these challenges have been modifications to the shape of the excitation light. Classical laser-based imaging approaches utilize a gaussian excitation beam which is focused to a spot or a sheet and scanned as required to excite the sample. As an alternative approach, bessel beams have been combined to introduce a structured pattern to the beam profile. The resulting 'lattice' of light has many benefits for live imaging. The most notable are a reduction of light exposure due to significant improvement in signal to noise while maintaining high resolution and optical sectioning. It is now possible to acquire data at previously unreachable combinations of acquisition speed and resolution with minimized phototoxic damage. This talk will outline the most recent implementations of lattice illumination and discuss how they are paving the way for the next generation of imaging solutions for a new era of life science exploration.

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