This article has been reviewed according to Science X's editorial process and policies. Editors have highlighted the following attributes while ensuring the content's credibility: Light sheet microscopy allows biologists to selectively light up a thin slice of a sample. This method can improve image contrast by reducing background light while remaining gentle on the sample, reducing photodamage and photobleaching.
Rice University's Anna-Karin Gustavsson has published a method in Nano Letters that expands the use of light sheet microscopy to work with a single objective in commercially available sample chambers. "Prior to this, light sheet microscopy typically required two objectives or specialized sample chambers," said Gustavsson, the corresponding author and an assistant professor of chemistry. "This new method allows us to use light sheet microscopy with a single objective in most commercially available sample chambers." Gustavsson started by adapting light sheet microscopy to work in microfluidic chips, which have tiny channels where researchers can study cells under varying conditions.
In that adaptation, her research team developed a single-objective approach in which the light sheet was reflected into the sample inside the microfluidic chip. "We realized we could 3D nanoprint a noncytotoxic insert to generate a mirror for light sheet reflection," said Nahima Saliba, co-first author and Rice alumna. "Then we could use this approach for single-objective light sheet imaging to reduce background fluorescence, photobleaching and photodamage directly inside the chip." The team wanted to extend the single-objective method beyond the microfluidic chips, which can be more complicated to work with and are not compatible with all samples.
The trick was to develop a pipeline for fabricating inserts that can fit snugly into commercially available sample chambers. "With our new approach, we can now place a combined insert into most types of sample chambers, add the cells and continue on with the assay as usual, growing and treating the cells for our experiments. When we are ready to image, the mirror allows us to create and manipulate the light sheet from the same objective that we use to detect the light from the sample," said Siyang Cheng, co-first author on the paper and a graduate student.
"Using our single-objective light sheet reduces background light when imaging, making it easier to see the individual molecules you're interested in. It also reduces harm caused by the light and photobleaching." Because the insert was 3D nanoprinted, inserts can easily be designed and printed for the most commonly available sample chambers. In fact, Gustavsson's lab has already created and provided open access to CAD files containing designs for multiple commonly used sample chambers.
"This opens up a more refined version of light sheet microscopy to anyone whose system would benefit from this type of selective illumination, enabling better imaging with less damage to the sample without having to adjust sample preparation workflows," Gustavsson said. Nahima Saliba et al, Versatile and Scalable Reflective Micromirrors for Single-Objective Light Sheet Microscopy, Nano Letters (2026). DOI: 10.1021/acs.nanolett.6c01709 BSc Life Sciences & Ecology.
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