Solicitation 75N98026Q00313 Q and A.pdf
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- Solicitation number
- 75N98026Q00313
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This is a Questions and Answers document for federal solicitation 75N98026Q00313 addressing clarifications on a histopathology project involving human brain tissue samples.
The project involves processing 58 human brain autopsy large-format tissue samples containing multiple anatomically distinct Central Nervous System regions. Each case will yield 16 tissue blocks (928 total blocks), with all blocks receiving H&E staining (928 H&E slides total) and 30 immunohistochemical (IHC) stains performed on select blocks across the 58 cases (1,740 IHC slides total). Staining will be conducted using biotin-free assays with four government-provided antibodies: Phospho-TDP-42 clone 11-9 (Cosmo Bio), Phospho-Ser129 alpha-synuclein (Abcam), Beta-amyloid (Abcam), and Phospho-Tau (Ser202, Thr205) Monoclonal Antibody AT8 (Invitrogen). Samples will be mounted on standard 1x3" paraffin-embedded slides; gelatin processing is not acceptable as antibodies have not been validated on gelatin tissue. No pre-stain optimization is required. The government will provide all antibodies to the contractor.
Samples will be submitted on a rolling basis with a few cases sent weekly, expecting 10 business day turnaround from receipt to completion. No validated staining protocols currently exist for these antibodies and tissues. The contractor must maintain a minimum of 67TB of storage capacity with platform access for three users capable of viewing, annotating, taking snapshots, and downloading raw files. No specific image file format is required. There is no anticipated expansion beyond 58 samples, and current requirements are limited to technical laboratory services only, with no future image analysis or pathologist interpretation expected at this time.
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Q & A for Solicitation 75N98026Q00313
Q: The outline mentions:
'Chromogenic immunostaining on 30 slides per sample, using a biotin‐free assay:
o Phospho-TDP-42 clone 11-9, Cosmo Bio o Phospho-Ser129 alpha-synuclein, Abcam o Beta-amyloid, Abcam o Phospho-Tau (Ser202, Thr205) Monoclonal Antibody (AT8), Biotin, Invitrogen'
Would this be 30 slides for 58 samples (1740 total), or 30 slides x 4 antibodies x 58 samples total (6960 total)?
A: Confirming 30 slides for 58 samples (1740 total)
Q: In addition, does the end-user have validated staining protocols already established for these antibodies and their tissues?
A: No
Q: 1) Please confirm that we are using standard 1x3" slides for the project, and not 2x3" slides.
A: Yes. 1x3” slides confirmed.
Q: At what rate are we anticipated to receive the samples? Will all samples be sent at the same time, or at different times? If different times, what is the expected cadence? Having this information would be very important for the bid.
A: A few cases will be sent to the vendor each week, not all at once. We expect weekly pick-ups/deliveries with 10 business day turn-around.
Q: 3) Can you please confirm the counts of the requested project:
58 samples x 16 blocks/sample = 928 samples
A: Correct
Q: From these 928 blocks, we will take 30 levels, with each level including: (1) H&E, (1) p- TDP-42 IHC, (1) pS129 A-synuclein IHC, (1) Beta-amyloid IHC, (1) pTau (please note that the RFQ lists a biotinylated version but that is neither necessary nor recommended)
A: This is not correct. For each block and H&E is required. 928 total H&E stained slides.
Immunostains will be performed on select blocks for a total of 30 immunostains per case (58 x 30 = 1740 IHC stained slides).
Q: Thus, the expectation is, in total, for the preparation of:
928 blocks x 30 levels/block = 27,840 H&E
928 blocks x 30 levels/block = 27,840 p-TDP-42 IHC
928 blocks x 30 levels/block = 27,840 pS129 A-synuclein IHC
928 blocks x 30 levels/block = 27,840 Beta-amyloid IHC
928 blocks x 30 levels/block = 27,840 pTau IHC
For a total of 27,840 slides x 5 stains = 139,200 slides (= 139,200 WSI)
A: This is not correct. The correct numbers are:
58 cases X 16 blocks = 928 blocks to trim and prep
58 cases X 16 blocks X 1 H&E = 928 H&Es
58 cases x 30 IHC on select blocks = 1740 IHC stained slides
Q: Quote specifies paraffin but we process in gelatin, which is a better option. Is that okay?
A: Paraffin is required for this project as the antibodies have not been validated on gelatin
Q: What/how many antibodies are being requested?
A: Government will provide these antibodies to the Contractor:
• Phospho-TDP-42 clone 11-9, Cosmo Bio
• Phospho-Ser129 alpha-synuclein, Abcam
• Beta-amyloid, Abcam
• Phospho-Tau (Ser202, Thr205) Monoclonal Antibody (AT8), Biotin, Invitrogen
Q: What specifically are the “58 large multi-piece brain tissue samples”
A: These are human brain autopsy large-format tissue samples containing many anatomically distinct Central Nervous System (CNS) regions per case.
Q: 5-10 business days is likely not realistic to perform embedding, sectioning , staining and imaging for anyone. However this depends on answering the details above about the brains, # of antibodies, etc. to understand size of project.
A: This turnaround time of 10 days is reasonable, especially since the samples will not be sent to the Contractor all at once. The aim will be rolling submissions by the Government to the Contractor with a few samples each week.
Q: Would the staining benefit from a pre-stain optimization of the antibodies on these tissues (additional charge option)?
A: No.
Q: Are there preferred image file formats required by NIH?
A: No. However, it is necessary to have a minimum of 67TB of storage on vendor’s platform, along with 3 users with ability to access to view, annotate, snapshot, and download raw files.
Q: Will there be any anticipated expansion beyond 58 samples?
A: No.
Q: Are there future expectations for image analysis, pathologist interpretation, or only technical laboratory services?
A: Not at this time
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