Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling
August 19th, 2017
This video article provides a detailed demonstration of the procedures required to successfully remove the pancreas from a mouse by dissection for histological analysis and metabolic profiling.
The primary goal of this video article is to provide detailed visual instructions outlining the dissection of a mouse pancreas. The instructions should be useful to researchers who study mouse models of human pancreatic diseases and who need to harvest the mouse pancreas by dissection for histological analysis or metabolic profiling. The video article also provides detailed instructions to perform a terminal blood collection using a heart puncture procedure.
Displayed is a operating table with the needed supplies. These include one glass jar with lid, one 15 mL falcon tube, one pair of surgical scissors, one squeeze bottle of 70%ethanol, two foam boards, two forceps, two one mL 21 gauge syringes, two 50 mL falcon tubes, one centrifuge tube, one cryo-vial, four surgical pads, ten pins, a dispenser of sterilizing wipes and a sharps container. Displayed is the post-operating table with the needed supplies.
These include one analytical balance, one four liter Dewar of liquid nitrogen, a shallow wide mouth Dewar, a floating microtube rack, one pair of surgical scissors, two forceps, five cryo-vials and a dispenser of sterile wipes. To perform euthanasia, the mouse is first placed in the glass jar containing a gauze pad soaked with a few drops of isoflurane, and then covered with the lid until the mouse becomes unconscious, which typically takes about one minute. The mouse is then removed from the glass jar and a foot pinch test is performed to ensure that the mouse cannot experience discomfort.
If the foot pinch test fails, the mouse is returned to the glass jar and the lid replaced until the foot pinch test can be performed successfully. Place the mouse on its back and pin the limbs of the mouse to the surgical foam board and wet the ventral side of the mouse with 70%ethanol. Pinch the fur near the urethral opening with the forceps and slightly pull upward.
Make an incision through the abdominal cavity, starting from the urethral opening, up the midline and ending at the chin. Near the starting point of the initial incision, grab one side of the fur and make another incision downward and diagonal toward the back paw. Repeat this in the same manner on the opposite side.
Locate the heart to perform the terminal heart puncture. To do this, grasp the sternum with the forceps and cut into the diaphragm. Quickly cut upward through the ribcage and remove the cut portion to expose the beating heart.
Once the heart is located, remove the pericardium and perform the terminal heart puncture with the syringe. For optimal blood collection, use the plunger of the needle to mimic the pumping action of the heart and avoid drawing too quickly. Dispel the blood into the centrifuge tube and dispose of the syringe into the sharps container.
After the heart puncture is performed, euthanasia is carried out by removing the attachments connecting the heart. Locate the stomach on the left side of the mouse. The pancreas has the appearance of a diffuse lighter color tissue that is attached to the stomach and duodenum and all along the small and large intestines and is attached to the spleen.
Begin to gently separate the pancreas from the stomach and duodenum by using the forceps. Continue to separate the pancreas from the small intestine continuing from the jejunum and ileum sections and finally to the cecum and large intestine. At the cecum, reposition the forceps and continue separation of the pancreas along the remaining colon towards the rectum.
At this point, it is convenient to cut and remove the section from the stomach to the region of the colon immediately preceding the rectum. Locate the pancreas and attached spleen. Slide the pancreas towards the right side of the mouse.
Cut the remaining connections between the pancreas and thoracic cavity to fully detach the pancreas and adjoined spleen. Remove the pancreas and spread it out to examine. If desired, other organs may be removed at this point.
After extraction, the samples are moved to the post operative area. Each organ will be weighed and then placed into their respective cryo-vial. Record any observed irregularities.
Once the organs are in cryo-vials, place them into the floating microtube rack and then into the Dewar with liquid nitrogen. After snap freezing, the organ should be stored at minus 80 degrees Celsius for long-term storage. All of the dissection tools used should be sanitized using the sterilizing wipes.
The falcon tube lined with the isoflurane soaked surgical pad should be capped. The surgical pad on the foam board should be replaced with a fresh surgical pad. Portions of the mouse that were not collected should be disposed of according to the facility's animal disposal policy.
The following items should be handled accordingly. Once these items are properly stored, a multitude of experiments can be conducted and include, but are not limited to, fluorescence microscopy, hematoxylin and eosin histology, immunohistochemistry, mass spectrometry, and nuclear magnetic resonance spectroscopy. With the pancreas being a primary organ for metabolic activity and insulin production, this procedure allows for the preservation of the physiological characteristics.
By isolating the pancreas, future analysis may be conducted on the sample. This procedure allows for the comparison and study of interactions from other tissues within the same organism, within the same timeframe.
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Overview
This video article provides detailed visual instructions for the dissection of a mouse pancreas, aimed at researchers studying pancreatic diseases. It includes procedures for harvesting the pancreas for histological analysis and metabolic profiling, as well as terminal blood collection via heart puncture.
Key Study Components
Area of Science
- Neuroscience
- Biology
- Histology
Background
- Mouse models are essential for studying human pancreatic diseases.
- Dissection techniques are critical for accurate tissue collection.
- Histological analysis and metabolic profiling provide insights into disease mechanisms.
- Terminal blood collection is necessary for comprehensive metabolic studies.
Purpose of Study
- To demonstrate the dissection of the mouse pancreas.
- To provide researchers with a reliable method for tissue harvesting.
- To facilitate the study of pancreatic diseases through detailed procedural guidance.
Methods Used
- Dissection of the mouse pancreas using surgical instruments.
- Terminal blood collection via heart puncture.
- Preparation of necessary supplies for the dissection process.
- Visual instructions to enhance procedural understanding.
Main Results
- Successful dissection of the pancreas demonstrated.
- Clear visual guidance provided for researchers.
- Effective methods for terminal blood collection outlined.
- Comprehensive overview of required materials and procedures.
Conclusions
- The video article serves as a valuable resource for researchers.
- Accurate dissection techniques are crucial for metabolic studies.
- Visual aids enhance the learning experience for complex procedures.
Frequently Asked Questions
Executive Industry Relevance
Accurate dissection of the mouse pancreas enables reliable tissue harvesting for downstream metabolic profiling and histological analysis in pancreatic cancer research. This procedural standardization supports target validation and biomarker discovery by ensuring sample integrity and reducing variability in preclinical models. The method enhances predictive confidence in early discovery workflows by providing a reproducible foundation for mechanistic studies of disease progression.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Scientific Value: Enables interrogation of therapeutic hypotheses through precise isolation of pancreatic tissue for metabolic and histological evaluation.
- Operational Value: Supports biological de-risking by providing consistent tissue samples for target engagement and pathway analysis.
Screening & Assay Development
- Scientific Value: Prepares validated biological systems for assay standardization by ensuring intact pancreas extraction for metabolic readouts.
- Operational Value: Addresses reproducibility requirements through standardized dissection techniques that minimize pre-analytical variability.
Translational & Preclinical Research
- Scientific Value: Maintains disease relevance by preserving pancreatic tissue architecture for staging cancer progression in preclinical models.
- Operational Value: Enables continuity from discovery through preclinical validation by providing harvest-ready tissue for multi-omic analysis.
Pipeline & Workflow Integration
The dissection method integrates into the discovery continuum by supplying high-quality pancreatic tissue for metabolic profiling and histological staging, directly supporting lead identification and preclinical validation stages.
- Discovery Biology: Facilitates hypothesis testing and pathway clarification by enabling collection of disease-relevant pancreatic tissue from genetically engineered models.
- Screening: Ensures assay readiness through standardized tissue preparation that supports consistent metabolic and histological outputs.
- Analytics: Generates quantitative measurements from tissue extracts that allow comparison of metabolic states across experimental conditions.
- Translational Research: Connects to preclinical continuity by preserving tissue for longitudinal analysis of cancer progression and treatment response.
- Enterprise Reuse: Establishes a reusable tissue harvesting capability applicable across multiple pancreatic cancer models and metabolic studies.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence by reducing mechanistic ambiguity through reliable tissue sampling for metabolic and histological correlation.
- Operational Value: Enhances standardization and scalability of pancreas harvest across research teams and experimental batches.
- Strategic Value: Improves go/no-go decisions by providing high-fidelity preclinical data that reduces late-stage biological risk in target validation.
- Portfolio Impact: Supports risk-adjusted prioritization by enabling accurate staging of pancreatic cancer progression in preclinical models.
Implementation Considerations
- Requires proficiency in mouse anatomy and dissection techniques to avoid tissue damage during pancreas isolation.
- Necessitates access to surgical instruments, sterilization supplies, and cryogenic storage for post-dissection sample preservation.
- Demands cross-team standardization of dissection protocols to ensure reproducibility across different operators and facilities.
- Involves adaptation considerations when applying the technique to different mouse strains or disease models with varying pancreatic morphology.
- Includes practical limitations related to the diffuse nature of the mouse pancreas, which requires careful separation from surrounding tissues to maintain sample integrity.
