A wide variety of technologies on the market allow scientists to image cells and tissues in 3-D. However, common techniques such as wide-field or confocal microscopy are often challenged by a certain degree of inefficiencies, photobleaching and photodamage to living tissues and cells, particularly when examining cells over long periods. Light sheet fluorescence microscopy (LSFM), also known as selective plain illumination microscopy (SPIM), reduces stress to living cells and tissues by focusing laser beams on only a single plane or sheet, minimizing light exposure and virtually eliminating the damaging effects of out-of-focus light found in wide field or confocal microscopy.

Light sheet microscopy technology has existed for decades. Through recent advances in technology, it has overcome many of its past hurdles, such as low resolution and slow acquisition speeds. Benefits of LSFM are many, the two biggest being the reduction of cellular damage and the drastic increase in analysis speed over other 3-D microscopy methods. However, commercialization of the technology is still in its infancy, with only a handful of vendors manufacturing off-the-shelf systems. The bulk of the installed systems are homemade, which is often a complex and expensive process.

Carl Zeiss is among the technology leaders with its Lightsheet Z.1 multiview fluorescence microscope. The system has a spectral detection range of 400–740 nm and can accommodate specimens from under 1 µm to 5 mm in size. Its ability to accommodate specimens of such a wide size range allows scientists to use it for multiple applications, from imaging 3-D cell culture to whole organisms, such as Drosophila, and a wide range of large embryos, from fish to mouse.

LaVision BioTec manufactures the Ultramicroscope, a LSFM capable of capturing 3-D images on both micro and macro scales, with working distances of up to 5,500 µm. The system can illuminate a plane 0.1–10 mm wide and 4–40 µm thick. It has a magnification range of 1.26x–12.6x and a field of view ranging from 0.7 mm to 7 mm.

Applied Scientific Instrumentation makes systems that can be mounted on inverted microscopes (ix73-diSPIM) or onto its Rapid Automated Modular Microscope (RAMM-diSPIM). The three configurations offered are the fixed-sheet, standard single-sided and double-sided systems. For the fixed-sheet system, the light sheet is stationary, while for the single-sided system, the sheet is illuminated from one side, with emission on the other side. The double-sided system includes illumination and emission from both sides of the sheet panel.

The market for LSFM is estimated to be less than $10 million and is forecast to grow double digits. Applications of LSFM continue to be explored, with strong potential for monitoring embryogenesis, cell and tissue development, and 3-D cell cultures as well as small organism research.

LSFM at a Glance:

Leading Suppliers

• Carl Zeiss

• LaVision BioTec

• Applied Scientific Instrumentation

Largest Markets

• Academia

• Government

• Biotechnology

Instrument Cost

• $80,000–$500,000