In this eBook, you’ll find resources that discuss methods for improved cell counting accuracy, including a comparison of trypan blue and AO/PI fluorescence viability dyes.
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 Fluorescence Cell Counters.
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.
Erythrosin B
Erythrosin B is a biosafe and non-toxic colorimetric dye that can be used as an alternative to Trypan Blue for cell counting and viability assessment. Evaluation of cell viability using Erythrosin B dye exclusion is based on a similar principle to Trypan Blue, that intact cell membranes exclude the polar molecules whereas compromised membranes do not. Live cells remain unstained while dead cells are stained and have a darker appearance.
CHO 293 Cells with Trypan Blue Viability Assay
Mouse Primary Cells with AO/PI Viability Assay
For Research Use Only
The CellDrop Automated Cell Counter counts cells without slides, which reduces plastic waste in science labs. But how many plastic slides has it actually saved, and how much plastic is that? Watch this video to find out!
Technical Note 231 highlights an application note from Miltenyi Biotec on optimizing nuclei preparation workflow for genomic applications.
The Erythrosin B apps on CellDrop Automated Cell Counters enable rapid automated cell counting and viability of cell suspensions stained with Erythrosin B.
During our Automated Hepatocyte Counting webinar, we conducted a live Q&A session with our applications team.
The CellDrop Automated Cell Counter offers a simple, reliable solution for automated hepatocyte counts. Learn more in this webinar.
