How can I prepare single cell or single nucleus suspensions using Singleron’s PythoN instruments?

Collected and prepared tissue is placed into a Singleron PythoN^® Series Dissociation Unit with the appropriate reagents. The PythoN i^TM or PythoN Junior^® combines mechanical and reagent-based dissociation to generate single cell or single nucleus suspensions. Processing parameters, including rotation speed, temperature, rotation duration and direction, and number of repetitions, are tailored to the sample type and downstream application to achieve efficient yet gentle processing. After the run, the suspension is filtered and washed manually before cell number and viability, or nuclei number and integrity, are assessed.

Singleron’s PythoN i (8 channels) and PythoN Junior (2 channels) tissue dissociation instruments prepare single cell or single nucleus suspensions by combining mechanical and reagent-based dissociation. The workflow has four steps:

1. Load the tissue. Place the tissue in a disposable Singleron PythoN® Series Dissociation Unit with the appropriate reagents.
2. Select and run a program. Select a preset program for the tissue type or customize the program to adapt to the experiment. The following parameters can be adjusted: rotation speed, temperature, rotation duration and direction, and number of repetitions. The parameters for each of the channels can be adjusted independently. This allows the user to process different tissue types in parallel or set different starting and ending time for each channel.
3. Filter and wash. After the run, filter and wash the single cell or single nucleus suspension.
4. Perform quality control. For single cell suspensions, count the cells and measure viability. For single nucleus suspensions, count the nuclei and assess their integrity.

Singleron’s technical support team can recommend a starting program for the tissue and species.