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Using CellDrop to Count Isolated Nuclei

Using CellDrop to Count Isolated Nuclei
Sponsored Content by DeNovix Inc.Mar 5 2021
The successful isolation of nuclei is an essential step in both ATAC-sequencing workflows and single cell RNA-sequencing. Successful library preparation requires users to ensure that all sample input is non-clustered and debris-free.
Dual fluorescence counting using the CellDrop™ FL Automated Cell Counter is a useful tool, able to distinguish unlysed intact cells from isolated nuclei.
Cluster analysis software provides an aggregation estimate to the user, with the resulting images allowing the user to verify that any debris has been effectively removed.
Counting nuclei with AO/PI stain
Acridine orange and propidium iodide (AO/PI) are utilized to confirm successful nuclei isolation. In conventional cell viability testing, the AO/PI dye combination will stain live cells so that these fluoresce green while staining dead cells so that these fluoresce red. ....

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Quantifying Isolated Nuclei from Mouse Brain Using the CellDrop™ FL Automated Cell Counter

Quantifying Isolated Nuclei from Mouse Brain Using the CellDrop™ FL Automated Cell Counter
Isolating nuclei is an essential step in both single nuclear RNA-sequencing and ATAC-sequencing workflows.
It is important to ensure that samples obtained during library preparation are non-clustered and debris-free, but nuclei isolated from tissue samples in areas like the brain are prone to high debris in comparison with samples obtained from cultured cells.
It is also important that nuclei are accurately counted for single cell analysis, even in instances where debris does not impact upon library preparation.
The CellDrop™ FL Automated Cell Counter’s dual fluorescence counting capabilities can clearly distinguish debris, nuclei and unlysed intact cells via acridine orange/propidium iodide (AO/PI) counting. ....

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