Microscope Objectives for Ultra-Deep Biological Imaging
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Two dedicated microscope objectives optimized for deep, high-resolution imaging of life science specimens up to 8mm beneath the surface have been introduced by Olympus. The new 10x and 25x objectives are designed for multiphoton imaging using clearing techniques such as CLARITY, SCALEVIEW and SeeDB, along with live-cell imaging and light sheet microscopy.
Both objectives, featuring super-long working distances of 8mm, are designed to boost the capability of multiphoton microscopy and allow scientists to look deeper than they ever could before. In the intact (whole) mouse brain, for instance, structures as deep as thalamus and hypothalamus 8mm beneath the surface can be imaged -- a longstanding goal within neurobiology. The microscope objectives also have many applications in cell and developmental biology. The new objectives join a full line of high-resolution optics compatible with industry-leading Olympus multiphoton imaging systems including the Fluoview FVMPE-RS and FV1200-MPE microscopes; they also are easily adapted for use with custom-built light sheet and other single plane illumination microscopes.
A high transmittance range of 400-1600nm, extending deep into the infrared spectrum, allows deeper imaging with minimal damage to tissue and can be employed to combine conventional multicolor two- and three-photon imaging with label-free methods such as third- and second-harmonic generation. The objectives also feature a correction collar with a wide range of adjustment throughout the working distance of the lens. This is important because lenses that have no correction collar, or that have correction collars with more limited capability to adjust, cannot focus as well when working deep below the surface of tissue. A field number of 27.5 provides excellent scattered-light collecting abilities and a large field of view (1.8mm at 10x) enhances data collection.
Finally, the objectives can handle immersion in water, clearing reagents, silicone oil, and more. As a result, researchers can capture the sharpest, brightest, most detailed images and movies that have ever been captured of contiguous structures up to 8mm or more beneath the surface. This is several times deeper than is possible with other types of objectives. The 10x objective offers refractive index (RI) immersion capability from 1.33-1.52, and the 25x offers RI immersion capability of 1.41-1.52.
