Automation of Lysate Preparation

Automating FIA sample prep involves using specialized technologies and systems to streamline the process:

• Sample Loading: Samples (cells, tissues, or fluids) are automatically loaded into processing wells or tubes by robotic arms or liquid handlers. This step ensures precise and contamination-free handling.
• Reagent Dispensing: Lysis buffer is automatically dispensed into the sample using precision liquid handling. The reagent type and volume are controlled to match the sample type and desired output.
• Cell Lysis: The system performs chemical, thermal, or mechanical lysis (e.g., shaking, sonication) to break open the cells. Conditions are tightly regulated for consistent lysis across samples.
• Mixing and Incubation: Samples are mixed and incubated under controlled conditions (time, temperature) to ensure complete lysis and protein release. Shaking or vortexing can be automated for uniformity.
• Mixing and Clarification: Automated centrifugation or filtration removes cell debris, leaving a clear lysate. This step prepares the sample for downstream analysis like FIA.
• Transfer and Storage:Processed lysates are transferred to fresh tubes or microplates and stored or sent to the next automated module. Barcoding ensures proper tracking and documentation.

Automate Lysate Preparation with Opentrons:

Interested in automating the reagent dispensing steps in your lab’s Lysate prep workflows? Enter the Opentrons Flex, a walk up, no-code pipetting robot that scales with your lab. Let us handle the pipetting while you spend more time on science.

Benefits of Automation Over Manual Methods

• Increased Speed:Automated systems significantly speed up the sample preparation process, allowing for higher throughput and faster turnaround times, which is especially valuable in large-scale studies.
• Improved Reproducibility: Automation ensures consistent sample preparation by minimizing human error and variability between different samples or operators, leading to more reliable results.
• Higher Throughput: Automation allows for handling larger sample volumes simultaneously, ideal for high-throughput projects.
• Better Accuracy and Precision: Robots perform tasks with higher precision, reducing contamination and errors.
• Cost Savings: Though the upfront cost can be higher than manual methods, automation reduces labor and reagent waste in the long run.

Methods of Lysate Preparation

Chemical Lysis

Uses detergents (like SDS or Triton X-100) or chaotropic agents to break down cell membranes. It’s simple, scalable, and compatible with many downstream assays.

Mechanical Disruption

Applies physical force (e.g., bead beating, homogenization, or sonication) to break cells open. Ideal for tough samples like tissues or bacteria with rigid cell walls.

Freeze-thaw Cycles

Repeatedly freezing and thawing cells causes ice crystals to rupture membranes. It’s gentle and effective for soft cells but may be slower and less efficient.

Enzymatic Lysis

Uses enzymes like lysozyme or proteinase K to digest cell walls or membranes. Often combined with other methods for added efficiency and specificity.