Immunostaining Services

Overview of Immunostaining in Neuroscience Research

Introduction of Immunostaining Services

Immunostaining is a standard technique for the detection of specific proteins using antibodies. As the essential methods in numerous branches of scientific research, they have been widely used in various preclinical and clinical applications. These methods have been expanding and optimizing to further increase the scope of their use and improve their accuracy and reliability.

Types of Immunostaining

• Immunohistochemistry (IHC)

IHC is a variant of immunostaining and has been the most applied immunostaining technique. Before the straining process, the tissue or cells are preserved through fixation to maintain cell morphology and tissue architecture. To obtain high-quality immunostaining, it is necessary to overcome specific and non-specific backgrounds.

• Flow Cytometry (FC)

The flow cytometer can be used for the quantitative analysis of stained cell products in the solution to determine the proportion and level of cells expressing specific markers. It is also possible to exclude dead cell material and analyze specific cell populations according to the size and granularity of the cells. Compared with IHC, FC presents improved sensitivity and can be analyzed in multiple colors to measure multiple antigens at the same time.

• Western Blot (WB)

WB is a conventional molecular biology method that can be used for semi-quantitative analysis of expression levels of target proteins. The cells to be analyzed are lysed and the resulting proteins are electrophoresed on a gel to separate by size. Then the separated proteins are transferred to a membrane to be immunostained.

• Enzyme-linked Immunosorption Assays (ELISA)

The ELISA is a more effective method for semi-quantitatively or quantitatively determining protein expression levels. Proteins in solution are adsorbed to ELISA plates through nonspecific binding or capture antibodies. The detection methods always include colorimetric and chemiluminescence.

• Immuno-electron Microscopy

Electron microscopy (EM) is suitable for the study of the detailed microarchitecture of cells or tissues. EM allows the detection of specific proteins in ultra-thin tissue sections and the subcellular localization of proteins

Fig.1 Micrograph of a GFAP immunostained section of a brain tumor.

Immunostaining in Neuroscience

Neurons are principal cells of the nervous system, and the neuronal diversity makes the cells study extremely challenging. To better understand the human nervous system, researchers focus on neural tissues, neural-specific protein targets, and neural cell markers. The identification of neuronal cells using immunostaining allows the study of signaling pathways and the elucidation of disease mechanisms. Subsequent two-dimensional imaging and 3D imaging can better capture neuronal features such as structure and dynamic direction.

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

As a leading biotechnology company, we provide comprehensive immunostaining services tailored specifically for neuroscience research. Our services are designed to facilitate the detailed study of neural tissues, enabling researchers to gain deeper insights into the structure, function, and pathology of the nervous system. All our services are strictly for research use only.

• Consultation: Initial consultation to discuss project goals, tissue types, and specific requirements.
• Sample Submission: Guidance on sample collection, preservation, and shipping.
• Project Execution: Regular updates and communication throughout the project, with interim results if required.
• Final Delivery: Comprehensive report including high-resolution images, quantitative analysis, and detailed methodology.

Our immunostaining services are designed to support cutting-edge neuroscience research, providing reliable and insightful data to advance our understanding of the nervous system. We also offer flexibility in our services, including but not limited to:

Published Data

Nicholas M. Kanaan, et al. used a variety of rigorously validated tau-specific monoclonal antibodies with epitopes targeting different regions of the tau protein to further elucidate the localization of tau in adult CNS neurons and glial cells under physiological conditions.

In this study, immunostaining was performed in juvenile rat and monkey brain tissue. They determined physiological concentrations of tau in the hippocampus by quantitative sandwich ELISA, and assessed the cellular localization of tau using a variety of tau-specific antibodies against epitopes in different regions. It was clearly demonstrated that tau is located in the somatic dendritic and axonal regions as well as in the nuclei of some neurons.

Fig. 3 Tau distribution in the adult rat hippocampus.³

Applications

Some of the key applications of our immunostaining services in neuroscience include:

• Localization of specific proteins: Immunostaining allows researchers to identify the presence and distribution of specific proteins within the brain and nervous system.
• Examination of cellular and subcellular structures: Immunostaining can be used to visualize the morphology and organization of neurons, glial cells, and other cell types in the brain.
• Analysis of neuroinflammation and neurodegeneration: Immunostaining can be used to detect markers of inflammation and neurodegeneration in the brain, such as activated microglia, astrocytes, and neuronal markers.
• Assessment of neuronal connectivity: Immunostaining can be used to label specific neuronal populations and visualize their connections within the brain.

By enabling the visualization and localization of specific proteins, this technique can help advance our understanding of neurological processes and ultimately lead to the development of new treatments for neurological disorders.

FAQs

Q: What types of samples can you process for immunostaining in neuroscience research?

A: We can process a wide range of samples for immunostaining, including brain tissue sections, cultured neurons, brain slices, and whole mounts. Our services are designed to accommodate both fixed and fresh-frozen tissues. We ensure that each sample type is handled with the utmost care to preserve tissue integrity and antigenicity, providing you with high-quality, reliable results.

Q: What specific markers can you detect using your immunostaining services?

A: Our immunostaining services can detect a variety of markers pertinent to neuroscience research, including but not limited to neuronal markers (e.g., NeuN, MAP2), glial markers (e.g., GFAP, Iba1), synaptic markers (e.g., synaptophysin, PSD95), and disease-related proteins (e.g., amyloid-beta, tau). We work with both commercially available antibodies and custom antibodies provided by clients to ensure the specificity and relevance of the markers used.

Q: Do you offer multiplex immunostaining services?

A: Yes, we offer multiplex immunostaining services, allowing the simultaneous detection of multiple markers within the same tissue sample. This approach is highly beneficial for studying complex cellular interactions and co-localization of different proteins. Our multiplex protocols are carefully optimized to prevent cross-reactivity and ensure clear, distinct signals for each marker.

Q: What types of imaging techniques do you use to visualize immunostaining results?

A: We utilize a variety of advanced imaging techniques to visualize immunostaining results, including brightfield microscopy, fluorescence microscopy, and confocal microscopy. These techniques allow us to capture high-resolution images and provide detailed visualization of staining patterns. Additionally, we offer image analysis services to quantify and analyze the staining data accurately.

Scientific Resources

Neuronal Marker

Reference

1. Kanaan, Nicholas M., and Tessa Grabinski. "Neuronal and glial distribution of tau protein in the adult rat and monkey." Frontiers in Molecular Neuroscience 14 (2021): 607303.

For Research Use Only. Not For Clinical Use.