Multi-color flow cytometric analysis of samples with low cell numbers
AssigneeBecton, Dickinson and Company
InventorsAlbert Mach, Smita Ghanekar, Marja Suni
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Abstract
Aspects of the present disclosure include methods for processing a biological sample. Methods according to certain embodiments include contacting a biological sample having cells with an assay reagent that includes one or more analyte-specific binding members to produce a biological sample assay composition and introducing the biological assay composition into an inlet of a flow cytometer having an integrated acoustic separator. Systems, including a flow cytometer with integrated acoustic separator having one or more acoustic concentrator devices suitable for practicing the subject methods are also described.
Claims23
A method comprising:
- contacting a biological sample derived from a tumor comprising cells, cellular debris, tissue matter, cell aggregates and non-cellular components comprising connective tissue, free lipids, proteins and nucleic acid fragments with an amount of assay reagent comprising one or more analyte-specific binding members to produce a biological sample assay composition comprising cells labelled with an analyte-specific binding member and excess assay reagent;
- introducing the biological sample assay composition into an inlet of a flow channel of an acoustic separator that is integrated into a flow cytometer, wherein the biological sample assay composition is introduced directly following preparation into the flow cytometer without intervening steps and wherein the integrated acoustic separator comprises:
- a flow channel comprising an inlet and an outlet and walls that extend from the inlet to the outlet;
- and
- an acoustic field generator positioned adjacent to the flow channel and configured to produce an acoustic field in the flow channel;
- conveying the biological sample assay composition along the walls of the flow channel of the acoustic separator with a laminating wash buffer flowing therebetween;
- and
- applying an acoustic standing wave in the flow channel in a manner sufficient to move one or more of the cellular debris, tissue matter, cell aggregates, non-cellular components and excess assay reagent from the biological sample assay composition flowing along the walls of the flow channel into the laminating wash buffer flowing therebetween, thereby producing a waste fluid.
Dependencies are inferred from the claim text.