Single-Objective Light Sheet Microscopy at Rice University

Commercial cell chamber with an insert in one of the wells.
Photo Credit: Rice University/Jorge Vida

Scientific Frontline: Extended "At a Glance" Summary: Single-Objective Light Sheet Microscopy

The Core Concept: A novel microscopy technique utilizing a 3D-nanoprinted, noncytotoxic insert within standard sample chambers to reflect a light sheet into a specimen, enabling high-contrast imaging using only one objective lens for both illumination and detection.

Key Distinction/Mechanism: Traditional light sheet microscopy requires either two separate objective lenses or specialized sample chambers, which limits usability. This method uses a custom mirror insert to direct the light sheet, drastically reducing background fluorescence, photobleaching, and photodamage while allowing researchers to use commercially available sample wells and maintain standard cell-preparation workflows.

Origin/History: Developed in 2026 by researchers led by Anna-Karin Gustavsson at Rice University and published in Nano Letters. The method evolved from their earlier work adapting single-objective light sheet imaging for complex microfluidic chips.

Major Frameworks/Components:

• Light Sheet Microscopy: A method of selectively illuminating a thin slice of a sample to improve contrast and reduce damage.
• 3D Nanoprinting: Used to fabricate the customized, noncytotoxic mirror inserts that enable the single-objective approach.
• Commercial Sample Chambers: Standard multi-well plates and chambers used in biological assays, now compatible with light sheet imaging via the new inserts.

Branch of Science: Microscopy, Biophysics, Cell Biology, and Nanotechnology.

Future Application: The open-access CAD files for the inserts allow broad adoption across biological research, enabling high-resolution, low-damage imaging of living cells and delicate molecules without requiring complex, specialized equipment or altered sample preparation.

Why It Matters: By eliminating the need for custom chambers and a second objective lens, this innovation makes advanced light sheet microscopy far more accessible, gentler on samples, and easier to integrate into standard biological experimental pipelines.

Nahima Saliba works on her imaging setup.
Photo Credit: Rice University/Jeff Fitlow

Light sheet microscopy allows biologists to selectively light up a thin slice of a sample. This method can drastically improve image contrast by reducing the amount of background light while being gentle, reducing photodamage and photobleaching within a sample. Recently, Rice University’s Anna-Karin Gustavsson published a method in Nano Letters expanding the use of light sheet microscopy to include 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 where 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.

Anna-Karin Gustavsson and Siyang Cheng in a microscopy room.
Photo Credit: Rice University/Jorge Vidal

“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 like normal, 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 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 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 for 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.

Funding: This work was supported by the National Institute of General Medical Sciences of the National Institutes of Health (R35GM155365) and startup funds from the Cancer Prevention and Research Institute of Texas (RR200025).

Published in journal: Nano Letters

Title: Versatile and Scalable Reflective Micromirrors for Single-Objective Light Sheet Microscopy

Authors: Nahima Saliba, Siyang Cheng, Prakash Joshi, and Anna-Karin Gustavsson

Source/Credit: Rice University | Rachel Leeson

Edited by: Scientific Frontline

Reference Number: biph100826_01

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