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Overview
This video protocol demonstrates an efficient technique for automated tissue dissociation of mouse spleens using the gentleMACS Dissociator. The method produces viable single-cell suspensions suitable for further applications.
Key Study Components
Area of Science
- Neuroscience
- Cell Biology
- Immunology
Background
- Tissue dissociation is crucial for obtaining single-cell suspensions.
- Manual dissociation can be tedious and inefficient.
- The gentleMACS Dissociator provides a reproducible and controlled method.
- Viable cell suspensions are essential for downstream applications.
Purpose of Study
- To demonstrate a time-saving technique for tissue dissociation.
- To prepare single-cell suspensions from mouse spleens.
- To enhance the efficiency of cell preparation for research.
Methods Used
- Use of gentleMACS Dissociator with specialized tubes.
- Preparation of buffer solution for tissue suspension.
- Filtration of dissociated tissue using Max Pres separation filters.
- Centrifugation to collect viable cell pellets.
Main Results
- Efficient dissociation of mouse spleens into single-cell suspensions.
- High percentage of viable cells demonstrated by flow cytometry.
- Successful removal of cell clumps for accurate analysis.
- Reproducibility of results across different sample sizes.
Conclusions
- The gentleMACS Dissociator significantly simplifies tissue dissociation.
- Proper buffer preparation is critical for successful outcomes.
- This method can be applied to various tissue types for cell analysis.
What is the gentleMACS Dissociator?
It is an automated device used for mechanical tissue dissociation to produce viable cell suspensions.
How do I prepare the buffer for dissociation?
Mix one part BSA stock solution with 20 parts AutoMax fencing solution containing EDTA.
What is the maximum volume of buffer I can use?
Do not exceed a total volume of 10 milliliters for the buffer.
How do I ensure sterility during the process?
Use the septum sealed opening of the C tube cap to pipette while maintaining sterility.
What is the significance of using Max Pres separation filters?
They help remove cell clumps, promoting successful separation and analysis of viable cells.
How can I confirm the viability of the cells?
Viability can be assessed using flow cytometry, staining dead cells with Propidium iodide.
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