Download this free resource to explore advances in quality control methods for single nuclei sample preparation.
Optimized for CellDrop™ Cell Counting Workflows
DeNovix CellDrop Assay Kits are simple to use and designed for Brightfield and Fluorescence applications preloaded on CellDrop instruments. DeNovix offers multiple assay kits for cell counting viability assessment, including Acridine Orange / Propidium Iodide (AO/PI) for the CellDrop FL Fluorescence Cell Counter.
AO-PI Viability Assay
The Acridine Orange (AO) / Propidium Iodide (PI) assay is used for cell counting and viability. AO is a nucleic acid-binding fluorophore that is cell membrane permeable and suitable for selective staining of nucleated cells. PI is a nucleic acid-binding dye that is impermeable to live cells and suitable for staining dead, nucleated cells. All live, nucleated cells fluoresce green due to AO, while dead, nucleated cells are stained with both AO and PI and fluoresce red.
Acridine Orange
The Acridine Orange (AO) Dye is used for total cell counting. AO is a nucleic acid-binding fluorophore that is cell membrane permeable and suitable for selective staining of nucleated cells.
Propidium Iodide
The Propidium Iodide (PI) Dye is used for viability assessment. PI is a nucleic acid-binding dye that is impermeable to live cells and suitable for staining dead, nucleated cells.
Yeast
The Fluorescein Diacetate (FDA) and Propidium Iodide (PI) assay is used for yeast cell viability testing. FDA is a cell viability probe that is membrane-permeable to live yeast cells. Inside a live cell, FDA is cleaved to give fluorescein by the action of intracellular esterases. Propidium Iodide dye is used for viability assessment. PI is a nucleic acid-binding dye that is impermeable to live cells and suitable for staining dead, nucleated cells. Live cells are reported by FDA in the green channel, dead cells by PI in the red channel.
Trypan Blue
The Trypan Blue (TB) dye is used for brightfield viability assessment. TB is an azo dye that is excluded from cells with intact cell membranes, only staining non-viable cells. The trypan blue assay is used to assess the number of viable cells present in a cell suspension.
CHO 293 Cells with Trypan Blue Viability Assay
Mouse Primary Cells with AO/PI Viability Assay
For Research Use Only
This technical note demonstrates Erythrosin B as an alternative to Trypan Blue as a cell viability stain using the DeNovix CellDrop.
During our Optimizing Nuclei Extraction & Counting for Single Cell Sequencing webinar, we conducted a live Q&A session with our applications team.
DeNovix instruments are compatible with eduroam using login credentials from your institution. Watch the video or follow the steps in this tech note to connect.
Explore recommended methods for optimizing nuclei counts before downstream sequencing applications, including the CellDrop Automated Cell Counter.
As nuclear transcriptomes mirror cell cytosolic profiles, nuclei isolated from brain tissue samples is a valuable tool for characterizing different brain cell types.
