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LICONN is a research method that combines light microscopy, physical tissue expansion and molecular labeling to map neuronal connections alongside selected molecules in the same brain sample. A 2025 report describes the technique enlarging mouse brain tissue 16-fold, making fine structures easier to image and trace while retaining labels that can help distinguish synapse types.
What LICONN measures
LICONN stands for light-microscopy-based connectomics. Its goal is to show how neurons connect while also identifying selected molecular features in the imaged tissue. That combination matters because a structural image may show where cells meet without clearly revealing which molecules are present at a given connection.
The method was reported by M. R. Tavakoli and colleagues in Nature in 2025. The account summarized here is Tim Wogan’s report for Chemistry World; the study concerns mouse brain tissue.
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Ordinary visible-light microscopy is constrained by diffraction. The Chemistry World report gives about 200 nm as the best-case resolution for conventional light microscopy, while some fine synaptic details are on the scale of tens of nanometres. The gap makes it difficult to resolve every relevant structure directly in an unexpanded sample.
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LICONN uses swellable hydrogel chemistry to enlarge tissue before light microscopy. In the study as described by Chemistry World, the protocol increased tissue size 16-fold. That figure describes the reported study, not a guaranteed expansion for every tissue or protocol. Enlarging the sample makes fine features more accessible to light microscopy; it does not mean that the microscope itself has acquired a universally higher native resolution.
What the reported workflow involves
The report describes a sequence of preparation and imaging steps rather than a ready-to-follow laboratory protocol:
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- A solution containing acrylamide monomer was delivered through the mouse vascular system alongside fixative.
- After the brains were removed and sliced, sodium acrylate was used to form a swellable hydrogel.
- The tissue was heated and treated with chemicals to disrupt its mechanical cohesiveness, then went through additional hydrogel steps that expanded it.
- Researchers applied antibodies to label selected molecules and imaged the expanded sample with light microscopy.
These details come from the secondary report. Exact concentrations, timings, equipment settings and reproducibility conditions should not be inferred from this summary; the paper and its supplementary methods are needed for a reproducible protocol.
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Why combine molecular labels with structural tracing?
Electron microscopy can reveal very fine structure, but by itself it provides little information about which molecules are present. Researchers can correlate electron microscopy with fluorescence microscopy to add molecular information, but the report describes that approach as requiring specialized equipment and expertise and as involving a resolution mismatch between imaging modalities.
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LICONN aims to combine fine structural tracing and molecular labeling within a light-microscopy workflow. The report gives inhibitory synapses as an example: they can be less conspicuous in structural images than excitatory synapses, so labels for selected molecules may help researchers identify them. A molecular label supports conclusions about the marker it detects; by itself, it does not establish synaptic function or physiology beyond that marker.
How the researchers traced neuronal connections
According to Chemistry World, the team manually traced neurons across multiple images and used the resulting traces as examples to train a deep-learning system. After three rounds of optimization, the model reportedly made reliable independent predictions. The report does not provide a quantitative accuracy figure here, so the result should not be read as a general performance guarantee for other samples, datasets or laboratories.
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How LICONN fits alongside other imaging approaches
| Approach | What it can contribute | Important qualification |
|---|---|---|
| Conventional light microscopy | Imaging of labeled tissue; the report gives about 200 nm as best-case diffraction-limited resolution. | That scale can make the finest synaptic details, reported at tens of nanometres, difficult to distinguish directly. |
| Electron microscopy | Fine structural detail. | Provides little information on which molecules are present without additional molecular-imaging methods. |
| Correlated electron and fluorescence microscopy | Can combine structural detail with molecular labeling. | The report says it requires specialized expertise and equipment and involves a resolution mismatch between modalities. |
| LICONN | Aims to combine neuronal-connection tracing and selected molecular labels using expanded tissue and light microscopy. | The cited account describes a mouse-brain method-development study, not routine clinical use or validated performance across tissues. |
The resolution figures and comparisons in this table are those reported by Chemistry World, rather than independent measurements of the study’s performance.
What the method does—and does not—establish
LICONN is a research technique, not a clinical imaging test or a consumer product. The report presents broader access for neuroscience laboratories and possible use in other organs as potential applications, not outcomes already established across labs, organs or human samples. Its reported result is a method for studying mouse brain tissue that brings structural tracing and molecular information together.
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Source: Tim Wogan, “Blown up brain cells’ connections and chemistry traced at incredible molecular resolution,” Chemistry World, 22 May 2025. The report cites M. R. Tavakoli et al., Nature (2025), DOI 10.1038/s41586-025-08985-1.
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