Preparing a viable single-cell suspension requires careful consideration of the liquid environment. The buffers and solutions used during mechanical or enzymatic tissue dissociation are critical for maintaining cell health and achieving a high yield of live cells. The choice of solution supports the entire process, whether performed manually or with a standardized tissue dissociator.
A Balanced Salt Foundation
Most procedures begin with a balanced salt solution as the base. These solutions, such as Phosphate Buffered Saline (PBS) or Hanks’ Balanced Salt Solution (HBSS), provide essential ions to maintain osmotic pressure and a stable pH. This foundation prevents cell rupture or shrinkage and creates a physiologically compatible environment for the cells during the tissue dissociation single cell isolation process.
Incorporating Enzymatic Activity
For many tissues, enzymes are added to the base solution to break down the extracellular matrix. Common choices include collagenase to digest collagen and trypsin to disrupt cell-to-cell adhesion. The specific enzyme blend and its concentration are tailored to the tissue type. A programmable tissue dissociator often uses pre-warmed enzyme solutions to ensure consistent activity throughout the run.
Protecting Cellular Integrity
To safeguard cells from enzyme over-digestion and mechanical stress, protective agents are frequently included. Serum albumin or specific enzyme inhibitors can be added to neutralize trypsin activity once the desired dissociation is achieved. Chelating agents like EDTA are also common, as they bind calcium ions to help separate cells held together by calcium-dependent adhesions.
Using the appropriate combination of these solutions is fundamental for successful tissue dissociation single cell workflows. The reagents and protocols compatible with BPLabLine‘s tissue dissociator systems are designed to support this careful balance, helping researchers obtain high-quality single-cell suspensions for downstream analysis.
