Multi-Immersion Objectives

Name: Multi-Immersion Objectives
Brand: Applied Scientific Instrumentation Inc.

ASI and Special Optics developed two objective lenses for light sheet imaging of cleared tissue samples as well as live cell imaging. Because of their unique curved first surface, both objectives work with all media regardless of refractive index — air, water, glycerol, oil, and cleared tissue media all work equally well. A common problem in live cell imaging is spherical aberrations as imaging goes deeper into the sample. This can be reduced by matching the medium refractive index to the sample refractive index.

However, there are residual aberrations from using e.g. a water dipping objective with non-water medium. This aberration is eliminated by using these objectives. The 54-10-12 objective has an NA of 0.4, 17x magnification, 12 mm working distance, and 1.2 mm field of view. The 54-12-8 has an NA of 0.7, 24x magnification, 10 mm working distance, and 1.0 mm field of view. The actual NA and magnification are both proportional to the immersion medium refractive index; stated values are for RI 1.45.

The long working distance and relatively slender profile of these objectives is critical for light sheet imaging, and stands in contrast to other cleared tissue objectives which, despite their long working distances, cannot be co-focused because of their wide barrels. When used in multi-view light sheet like the ASI diSPIM, the objectives offer 800 nm and 450 nm resolution respectively in X, Y, and Z in FocusClear (CLARITY) solution. The fluidic sealing has been engineered to allow the objective lenses to be dipped into harsh chemicals including DBE, BABB, ECi, CUBIC, and FocusClear.

Applications
     —   Dual-view cleared tissue imaging, e.g. ct-dSPIM
     —  Single-view cleared tissue imaging, e.g. COLM/OpenSPIM/ct-ASLM setups
     —   Live cell imaging with RI-matched media
     —   Any imaging requiring very long working distances with modest NA

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