Troubleshooting Guide: Cell Enrichment Columns

Our cell separation columns are designed as a rapid, one step tool to enrich for specific cell populations. We have attempted to make the columns simple and user friendly. To ensure that all users get maximal efficiency from these columns, we have summarized some common problems below. We attempt to identify the likely sources of these problems and possible courses of action to be taken to avoid them. We urge users to review these tips and if additional advice is required to contact our technical service department.

Additional Information:

Problem: Column buffer does not drip after removal of bottom cap

Possible Source Test or Action
Air lock in column tip Tap side of column to dislodge air bubble

Problem: Very slow flow rate following cell loading

Possible Source Test or Action
Cell clumps and debris have accumulated on the top white filter Remove cell debris that has accumulated on the white filter with a sterile pipette
Reduce the amount of time single cells incubate in a small volume as it may reduce the tendency of cells to form clumps

Problem: Cell recovery low

Possible Source Test or Action
Poor specimen preparation Ensure that suspension contains single cells with no clumps and minimal RBC
Column was either overloaded or underloaded Load the optimal number of total cells indicated in the product insert
Improper calculation Percent recovery should be calculated by dividing the number of target cells recovered by the number of target cells loaded

Problem: Cell purity is low

Possible Source Test or Action
Non-optimal number of total cells loaded Reduce the total number of cells loaded and determine by immunophenotype if the frequency of the contaminating population is reduced
Poor specimen preparation or tissue source High frequency of dead cells in recovered sample. Determine cell viability and perform immunophenotype analysis gating only on live cells