Recent years have seen anincreasing demand for cross-scale high-throughput imaging, however,conventional microscope objectives cannot simultaneously meet therequirements for large field and high-resolution imaging.

Mesoscale microscopeobjectives, with complex optical structures and excellent aberrationoptimization, can achieve both high numerical aperture andultra-large field of view (FOV), and significantly enhance theimaging throughput of optical microscopes.

Recently, a group led by SHIGuohua from Suzhou Institute of Biomedical Engineering and Technology(SIBET) designed a flat-field apochromatic objective lens structureunder a mesoscale field and developed a mesoscale microscopeobjective with the largest reported FOV and the widest working bandat sub-micron resolution.

Objective lens is the corecomponent of optical microscope, determining two critical parametersof microscopic imaging: resolution and imaging field of view (FOV).The resolution and FOV of the objective lens are interdependent. FOVof existing mesoscale objectives is concentrated between 3mm and 6mmdiameter.

This objective lens featuresan 8mm FOV diameter, a 0.5NA numerical aperture, and an imaging bandextending from 400-1000nm. Utilizing this objective lens for imagingmouse brain and kidney slices, researchers obtainedultra-high-throughput images with a single frame of 1.35 billionpixels (Figure 2a).

Current mesoscale objectivesare limited to a single imaging wavelength, capable of only visibleor near-infrared single-band imaging, and cannot meet therequirements for diverse fluorescence imaging.

The researchers constructed arelated imaging system (Figure 1) and achieved single and two-photonmesoscale imaging, for the first time, using the same objective lens(Figure 2b).

A quantitative comparison witha commercial 20x 0.5NA objective lens indicated that this mesoscaleobjective had similar imaging quality to the commercial lens butprovided an FOV over 40 times larger.

Experimental resultsdemonstrate that this objective lens has significant potential forlarge-scale sample high-resolution multi-wavelength imaging, such asbrain mapping, cross-brain region single and two-photon imaging, andhigh-resolution imaging of organoids.

Results of the study entitled“Large-field objective lens for multi-wavelength microscopy atmesoscale and submicron resolution” were published in the recentissue of Opto-ElectronicAdvances.

Figure 1.Mesoscopicobjective lens structure and imaging system configuration. (Image by SIBET)

Figure 2. Imaging results ofbiological samples. (Image by SIBET)

Contact

XIAO Xintong

Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences (http://www.sibet.cas.cn/)

Phone: 86-512-69588013 

E-mail: xiaoxt@sibet.ac.cn