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NeuroScience Associates

Frequently Asked Questions


General Questions

How long has NSA provided histology services?

NSA has served over 300 global research organizations since 1989.

What is the typical turnaround time for sectioning and staining?

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.

Can staining be done on adjacent sections after the initial project is complete?

Yes. All sections cut are collected creating a Resource of Sections for future staining.


Tissue Preparation Questions

Should I put the brains into a cryoprotect solution (e.g. sucrose) before shipping?

No. NSA uses a glycerol and DMSO formulation to protect against freeze artifacts.

Should the tissue be frozen to ship to NSA?

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.

If the brains I want processed by NSA are already frozen can they be processed successfully?

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.

What is the best method for shipping the specimens?

By overnight express carrier such as FedEx. See SHIPPING INSTRUCTIONS.


MultiBrain® Technology Questions

Can NSA arrange the specimens in the MultiBrain® Block to my specifications?

Yes. Indicate the plane and pattern you wish on the Brain/Section Layout Array diagram. Download and upload your arrays on this page.

How is the identity of each brain preserved in the MultiBrain® Block?

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.

Does NSA's MultiBrain® Matrix permeate the tissue as occurs with paraffin embedding?

No. The MultiBrain® Matrix encases the brains, therefore tissue staining is not affected.


Silver Degeneration Stain Questions

Can I execute the degeneration stain in my own lab?

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.

Is the time between injury/insult and when the animal is sacrificed important when assessing neurodegeneration?

Yes. There are optimal times for observing synaptic terminals, cell bodies and axons. See our reference section for more information.

Are silver stains all the same?

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.

Is FluoroJade as effective in staining degeneration as the silver degeneration stains?

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.

Does the Campbell-Switzer stain show the same features as the disintegrative degeneration 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.

How do the plaques revealed by the Campbell-Switzer stain in transgenic mouse models compare with staining using antibodies against different forms of amyloid?

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.


Antibody Questions

Can NSA do immunohistochemistry with antibodies not offered commercially?

Yes. The client can provide the antibody. NSA will then perform a titration series to determine the optimal dilution.

Is the section thickness of 30-40µ too thick for antibody staining?

No. By using TritonX-100, membranes are sufficiently fenestrated to allow antibodies access to the entire thickness of the section.

Does the MultiBrain® Matrix that surrounds the tissue interfere with antibody staining?

No. There is no interference by the matrix since it only encases, and does not infiltrate, the tissue.

Can sections be sent to clients so they can do their own staining?

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).


Cost Questions

What is the cost of the NSALabs® neurohistologic staining services?

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.


Contact Technical Support at NSA Labs via this form or call 865.675.2245 for shipping and preparation questions.