Organ Processing and Pathology Core

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Organ Processing and Pathology Core

• Martha Campbell-Thompson, DVM PhD, PI
• www.jdrfnpod.org/oppcservices.html

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Core Goals

• Serve as a central repository for all tissues
  procured through this Program (All organs/tissues
  from the OPOs will be shipped to this Core).
• Receive, process organs/tissues/serum/PBL from
  the OPOs, and distribute tissues to approved
  centers and investigators.
• Whole organ dissection for frozen and fixed
  samples.
• Frozen and paraffin serial section slides.
• Slide staining by HE and immunohistochemistry.
• Slide scanning and retrieval with web-based
  viewing (Aperio).

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Organization
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Core Functions
24/7 Procurement OPO ship tissues in UW Tissue
dissection Pancreas, Panc. LN, Spleen,
Non-panc. LN Isolated lymphocytes and
splenocytes OCT and paraffin blocks EM Space 400
sq ft
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Flow Chart for Organ/Tissue Dissection
Tissue Recovered from Donor in OR
OPPC Staff Notification by OPO
Tissue Received by OPPC Staff
Tissue Dissected by OPPC Staff
Store Cryotank
Pancreas Samples Cut (1cmx5mm) for Fresh and
Frozen
Tissue Sections for Freezing Placed into
Pre-labeled Cryo-vials
Tissue Sections for OCT Freezing Placed into
Pre-labeled Vinyl Molds
Tissue Sections for Fixation Placed into
Pre-labeled NBF Containers
30 40 Minute Timeline
Snap Freeze LN2
Samples submitted for processing and embedding
Add Tissue to Mold, Add OCT, and Drop in
LN2-cooled Isopentane for 2 minutes
Store -80C
Store -80C
Scan Slides
Archive Blocks and Slides
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Histology IHC

• Pancreas
• HE
• IHC
• Insulin double stains with Ki-67, CK-7,
• Cleaved caspase-3 Non-insulin cells
• Pancreatic and non-panc LN, spleen
• HE
• IHC CD3,4,8 B220 CD68
• Clinical grade antibodies

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nPOD Organ Trimming, Paraffin Blocks, and IHC
Workplan
Head
Body
Tail
A
C
D
Pancreata Blocks- head, body, and tail10
blocks 4 IHC/block Spleen Blocks- 3 LN Blocks-
4 7 blocks 5 IHC/block
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Data Processing Image Analysis
a Plt0.05 from no pancreatitis, ANOVA
Campbell-Thompson, et. al, unpublished
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Core Functions
Aperio ScanScope Virtual microscope on the Web
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Core Processes and Equipment
Tissue banking facility with approved IRB
protocols Equipment (-80 freezer on emergency
power, alarm system, back-up freezer cryotank
freezer bar coding labeling system, scanners,
computers) Good lab practice (GLP) compliant
histology Immunohistochemistry/ISH/FISH Dako
autostainer Microscopy Zeiss Axioskop with
image analysis software LIMS for submissions,
services rendered, ordering supplies, invoicing
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Laboratory Information Management System (LIMS)