Attachment 2 NIH NeuroBioBank Tissue Access Guidance

Guidance for Completing your NBB Tissue Request
Planning Your Tissue Request
 If you are not sure precisely what tissue, tissue regions, or quantities are best for your
research project, please submit a request for help to the NIH NeuroBioBank email
address before completing the on-line request.
 The Request Form requires you to provide a detailed rationale for 1) number of subjects
requested, 2) amount of tissue requested per subject, and 3) number of brain regions
requested. Please see the Acceptable Tissue Amount Guidelines (below) to plan your
request.
 The more restrictive your subject characteristics, the less likely a brain bank will have
tissue that meets your requirements. Ensure that any restrictions you place on subject
characteristics (e.g., postmortem interval (PMI), RNA Integrity Number (RIN), age range)
are necessary for your study to be valid.
 If you have not performed a pilot study in human tissue, your first request should be for
tissue to perform a pilot study using your proposed methods.
Completing Your NBB Tissue Request Form
1. Complete all applicable fields. Required fields must be completed in order for the system
to accept the request. Incomplete information will delay the review of your request,
especially Specimen Shipping Information.
2. Request Name should be a short descriptor.
3. Specimen Needs
a. This component is divided into 2 sections: Fixed Tissue Request and Frozen
Tissue Request. If you require both fixed and frozen tissues, please complete
both sections, and note in the “Subject Specimen requirements” box if the fixed
and frozen tissues need to come from the same subjects.
b. Each row should have one subject diagnostic category and one tissue type. For
example, if you require Brodmann Area (BA) 9/46 and BA21 from Unaffected
Control subjects and subjects with Major Depressive Disorder, the request would
have 4 rows.
c. “Unaffected Control” are subjects with no known psychiatric or neurological
diagnoses.
d. Please type out the name of the diagnostic category, and do not use
abbreviations.
e. Minimum Tissue Size must follow the Acceptable Tissue Amount Guidelines
(below). A detailed rationale must be provided for any deviations from these
guidelines.
4. Subject/Specimen requirements: this section should include any additional
requirements pertaining to the subjects/specimens themselves (e.g., PMI, RIN, fixation
type, sex, etc). As noted above, the more restrictive your subject characteristics, the less

likely a brain bank will have tissue that meets your requirements. Ensure that any
restrictions you place on subject characteristics are necessary for your study to be valid.
5. Request Details
a. Title of Research Plan that will appear at the top of your MTA.
b. Concisely describe the request according to the instructions. If you are
requesting multiple anatomical regions per subject, the rationale must be clearly
indicated here.
c. List each type of assay or method to be used with the requested tissue. Include
the amount of tissue needed for each assay/method per subject. For example,
“qPCR, 2mg grey matter per subject.”
d. Indicate whether you or your direct collaborators have used the method(s)
proposed in this request with human postmortem tissue before.
i. If the answer is “No,” then this request should be for tissue for pilot
studies.
e. Provide a rationale for the number of subjects requested. Acceptable rationales
include power analyses, or accepted standards in the field (with appropriate
citations).
Determining Your Tissue Needs
The following table indicates acceptable amounts of tissue that can be requested for different
types of studies. As different brain banks process and store their samples differently (see NBB
Best Practices), you may be supplied with more tissue than you request. However, each of the
tissue amounts below will provide enough material to perform each type of methodology listed.
These values were derived in collaboration with researchers who have successfully performed
each methodology using human brain tissue.
Method
FACS sorting with NeuN
HITS-CLIP
Immunoblot
Immunohistochemistry

Accepted Tissue Amount/Region/Subject
≤ 600mg of gray matter
≤ 300mg
≤ 10mg of tissue per protein examined
Sufficient tissue for ≤ 8 sections per condition
(May be provided as free floating sections,
formalin-fixed paraffin-embedded sections,
fixed tissue blocks, or paraffin-embedded
tissue blocks dependent upon region
requested and brain bank)

Mass Spec:
Simple Mass Spec with no enrichment or
multidimensional fractionation
Mass Spec with sucrose density gradient
based sub-cellular enrichment
Mass Spec with multidimensional
separation with or without isobaric
labeling tags (e.g., TMT or iTRAQ)
Co-IP with Mass Spec

≤ 15mg
≤ 350mg
≤ 150mg
50 – 500mg

Mass Spec for post-translational
modifications
Microarray
qPCR
RNASeq

50 – 500mg
≤ 5mg
≤ 5mg
≤ 60mg

If your methodology is not listed, or you require more tissue than is indicated, please provide a
detailed rationale supporting the amount of tissue you are requesting. Multiple brain regions
such as hippocampal subfields, and thalamic and hypothalamic nuclei are small, and many
investigators are interested in studying these regions. Ensure that your requests are
anatomically possible before submitting. If you need guidance on this issue, please contact the
NIH NeuroBioBank at neurobiobank@mail.nih.gov.