Frequently Asked Questions
NSA has served over 300 global research organizations since 1989.
Approximately 3-4 weeks. This depends on the number of stains and the total number of sections stained. For instance, NSA processes rodent brains in blocks of 16-25, with the capacity to generate over 10,000 sections per day. Capacity is rarely a consideration, rather, there are several steps each of which require a minimum number of days to complete to yield optimal results.
Yes. All sections cut are collected creating a Resource of Sections for future staining.
Tissue Preparation Questions
No. NSA uses a glycerol and DMSO formulation to protect against freeze artifacts.
NO! Do not freeze the specimens following fixation. This will likely create freeze artifacts. After perfusion, keep the brains or heads in fix, or later in buffer, and ship at room temperature. See - SHIPPING INSTRUCTIONS for specifics.
It is possible to prepare snap frozen tissues (that have not been fixed or cryoprotected) for MultiBrain®processing. If a tissue has not been cryoprotected (with sucrose or glycerol) there is the risk of freeze artifact (micro tears in the tissue, often rendering it useless for microscopy). Frozen brains should be immediately immersed in room temperature 10% phosphate buffered formalin. Thawing and initial fixation are accomplished in one step. After overnight in the formalin, switch the tissue to buffer and the tissue is ready to be shipped to NSA for processing with MultiBrain® Technology. The sections should reveal scant or no freeze artifact and the brains are typically able to be processed normally including a successful antibody staining procedure.
By overnight express carrier such as FedEx. See SHIPPING INSTRUCTIONS.
MultiBrain® Technology Questions
Yes. Indicate the plane and pattern you wish on the Brain/Section Layout Array diagram. Download and upload your arrays on this page.
At the time of embedding, only one brain at a time is outside of its container. Once the brain is in the matrix its position relative to the other brains is permanent. The unambiguous orientation of the MultiBrain® Array is provided by the black MultiBrain® Matrix Marker. One or more black dots is present in a designated corner of each MultiBrain® section.
No. The MultiBrain® Matrix encases the brains, therefore tissue staining is not affected.
Silver Degeneration Stain Questions
Yes. The foundation protocols are published, but it took NSA years to perfect our own variation and achieve consistent success and high quality in executing these stains.
Yes. There are optimal times for observing synaptic terminals, cell bodies and axons. See our reference section for more information.
No. There are numerous silver staining protocols, each tailored to reveal specific features. "Silver Stains" are as unique as any other stain in that they are each specifically designed to stain a particular feature. Silver simply refers to the staining agent (Silver) which stains the designated feature black. See: Switzer, R. C. III. Application of Silver Degeneration Stains to Neurotoxicity Testing. Toxicologic Pathology 28: 70-83, 2000.
No. Researchers are reporting that some forms of degeneration are unstained by FluoroJade. Also, as FluoroJade is a fluorescent stain, sections cannot be viewed practically at low magnifications as can be done with the amino cupric silver stain.
No. Each method reveals a different set of features. The Campbell-Switzer method reveals the neuritic plaques and tangles of Alzheimer's pathology. The disintegrative degeneration stain shows the products of neuronal disintegrative degeneration.
Some antibodies show only a subset of plaques revealed by the Campbell-Switzer method, while others appear to be a one to one match. The Campbell-Switzer stain is unique in its ability to differentiate its staining of immature (diffuse) and mature (congophilic) plaques, allowing researchers a unique observational capability.
Yes. The client can provide the antibody. NSA will then perform a titration series to determine the optimal dilution.
No. By using TritonX-100, membranes are sufficiently fenestrated to allow antibodies access to the entire thickness of the section.
No. There is no interference by the matrix since it only encases, and does not infiltrate, the tissue.
Yes. We can return free-floating sections for immunostaining or other staining. The client should inform NSA of the types of stains anticipated so the tissues are properly stored after sectioning, to optimize the success of the chosen stain(s).
The average cost per individual brain section is comparable to standard paraffin processing with one section per slide. With MultiBrain® sections, uniformity of staining across many brains is achieved, as well as fewer slides to store and faster analysis. Contact us for a custom quote. Current pricing is available upon request.
For shipping questions, please see our shipping information page.