Sparse-labeling Services

Overview of Sparse-labeling

Introduction of Sparse-labeling

Sparse-labeling means that the labeling density is about 1% and labeled neurons do not obscure each other. It is important for visualizing synaptic structures as well as following axons or dendrites across long distances.

In neuroscience, to clarify the developmental principles and functional logic of neural circuits, it is essential to access individual neurons. There are three main strategies to realize the analysis of a single nerve cell or its precursor, including sparse-labeling, the introduction of more labels, and improved image resolution. Among them, the labeling strategy still occupies a key advantage despite the current progress in nanoscale imaging. In general, sparse-labeling can be achieved by locating labels to specific cells or distributing them in a random manner or a combination of the two. Golgi straining remains a benchmark for sparse-labeling. Sparseness and randomness are the main characteristics of sparse-labeling and they are related because the labeling probability directly affects the labeling density. The sparseness of the labeling can be controlled by adjusting the random process.

Transcriptional Strategies for Sparse-labeling

It has been proved that compared with random labeling, the use of transgenic animals expressing reporters from cell-type specific promoter fragments is more efficient for many scientific problems. Some neuron types are sufficiently sparsely distributed for single-cell analysis, but spatial clustering neurons are more typical, and subdivision of the expression pattern is required. In this case, there are two types of transcriptional strategies, including deterministic sparse transcription and stochastic sparse transcription.

Fig.1 MORF3 Mice Enable Brainwide Sparse Labeling of Microglia and Astrocytes to Reveal Their Detailed Morphology.¹

Available Markers for Sparse-labeling

• Fluorescent proteins (FPs) - directly detectable and can be restricted to specific cell types.
• Chromogenic enzymes - with higher signal amplification.
• Small epitope tags - useful in fusion proteins.
• Synthetic dyes - can be sparsely distributed with DiOlistics.
• miniSOG - a small, genetically encodable protein module.

Available Methods for Sparse-labeling

• Golgi straining
• DiOlistics method
• Dye microinjections
• Low-titer viral infections
• MORF/Cre mouse lines

Creative Biolabs is one of the well-recognized experts who are professional in applying advanced platforms for a broad range of neurosciences research. We are pleased to use our extensive experience to offer the best service and the most qualified products to satisfy each demand from our customers. If you are interested in our services and products, please do not hesitate to contact us for more detailed information.

Services at Creative Biolabs

Our sparse-labeling services cater to neuron research and other biotechnology applications that require precise, targeted labeling of specific cell populations. With our sophisticated techniques and cutting-edge technology, we are able to selectively label individual cells or small groups of cells within a heterogeneous population. Our services offer several key features and benefits:

• Precision and accuracy: Our methods ensure that only the desired cell population is labeled, minimizing off-target effects and preserving the integrity of the experimental system.
• High-throughput capabilities: Our automated systems can process large numbers of samples with efficiency and consistency, providing reliable results for high-throughput screening and analysis.
• Flexibility and customization: We work closely with our clients to understand their research goals and experimental requirements, tailoring our labeling strategies to meet their specific needs.
• Comprehensive support: Our team of expert scientists and technical specialists are available to provide guidance and support at every step of the experimental process, from experimental design to data analysis.

In summary, our sparse labeling service provides a powerful tool for researchers in neuroscience. We also offer flexibility in our services, including but not limited to:

Published Data

Neurons are densely arranged in the brain and their dendrites and axons are intertwined with each other, making it impossible to see their morphology. Therefore, it is often necessary to use sparse-labeling methods to randomly label a small number of neurons to highlight their dendrites and axons.

Matthew B. Veldman et al. designed several Cre-dependent mouse strains for sparse-labeling, which confer Cre-dependent sparse-labeling based on single nucleotide repeat fiducials (MORFs) acting as random translation switches. Optimized MORF3 mice have membrane-bound multivalent immunoreporters that confer Cre-dependent sparse and bright labeling on thousands of neurons, astrocytes, or microglia in each brain, revealing their complex morphology. The MORF3 mice are suitable for his totransparent thick brain section imaging and immunoelectron microscopy.

Their study demonstrated a conceptually novel, simple, generalizable and scalable mouse genetic solution for sparse-labeling and elucidating the morphology of genetically defined neurons and glial cells in the mammalian brain.

Fig. 2 MORF3 mice enable brainwide sparse labeling of microglia and astrocytes.³

Applications of Sparse-labeling in Neuroscience

Our services are designed to assist neuroscientists in visualizing and studying specific populations of neurons within the brain with high precision and efficiency. By selectively labeling only a subset of neurons within a neural circuit, researchers are able to better understand the function and connectivity of those specific neurons without the noise and interference of neighboring cells.

• Researchers can target and label specific neuronal populations based on their genetic or anatomical characteristics, such as expression of a particular protein or location within a brain region.
• Researchers can track the activity of individual neurons, monitor synaptic connections, and manipulate neural circuits with unparalleled specificity.

We are committed to advancing neuroscience research through the development and application of innovative technologies for sparse-labeling of neurons.

FAQs

Q: What types of sparse-labeling techniques do you offer?

A: We offer a variety of sparse-labeling techniques including genetic methods, viral vectors, and chemical labeling. These methods can be tailored to specific research needs. Genetic methods involve the use of Cre-loxP or FLP-FRT systems, viral vectors can deliver genes selectively to target neurons, and chemical methods utilize fluorophores that selectively bind to specific cell types. Each technique is optimized for different experimental goals and model systems.

Q: Can your sparse-labeling services be used in conjunction with other neuroscience research techniques?

A: Absolutely. Our sparse-labeling services are designed to be compatible with a wide range of other neuroscience research techniques. These include electrophysiology, optogenetics, two-photon microscopy, and behavioral studies. Combining sparse-labeling with these techniques allows for a comprehensive analysis of neuronal function and connectivity, providing deeper insights into brain activity and its correlation with behavior and cognitive functions.

Q: What species or model organisms can be used with your sparse-labeling services?

A: Our sparse-labeling services can be applied to a variety of species and model organisms, including rodents (mice and rats), non-human primates, zebrafish, and Drosophila. We tailor our approaches based on the specific requirements of each model organism to ensure optimal labeling efficiency and specificity. This versatility allows researchers to study neural circuits across different species, enhancing the generalizability of their findings.

Q: What are the costs associated with your sparse-labeling services, and what factors influence the pricing?

A: The costs for our sparse-labeling services can vary based on several factors, including the complexity of the project, the type of labeling technique used, the model organism, and the scale of the experiment. We provide a detailed cost estimate after the initial consultation, taking into account all these variables. Our pricing is competitive, and we strive to offer the best value by delivering high-quality, reliable results. For a precise quote, we encourage researchers to contact us with their specific project details.

Scientific Resources

Morphology Research Methods in Neuroscience

Comparison of Various Viral Vectors in the Central Nervous System

References

1. Veldman, M.; et al. Brainwide genetic sparse cell labeling to illuminate the morphology of neurons and glia with cre-dependent MORF mice. Neuron. 2020, 108(1): 111-127. e6.
2. Jefferis, G.; et al. Sparse and combinatorial neuron labelling. Current opinion in neurobiology. 2012, 22(1): 101-110.
3. Veldman, Matthew B., et al. "Brainwide genetic sparse cell labeling to illuminate the morphology of neurons and glia with cre-dependent MORF mice." Neuron 108.1 (2020): 111-127.

For Research Use Only. Not For Clinical Use.