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Separating Bacteria by Capsule Amount Using a Discontinuous Density Gradient

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Published on: January 7, 2019

Dispersion serial dilution methods using the gradient diluter device.

Leslie Walling¹, Craig Schulz, Michael Johnson

¹Amgen, Inc., Thousand Oaks, California 91320, USA. l.walling@stratec.com

Assay and Drug Development Technologies

|February 28, 2012

Summary

This summary is machine-generated.

This study introduces the Gradient Diluter, a novel device that leverages dispersion for enhanced serial dilutions. This innovation expands dilution ranges to six orders of magnitude in small volumes, improving experimental efficiency.

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Area of Science:

Biochemistry
Analytical Chemistry
Biotechnology

Background:

Dispersion, a solute's dilution effect in liquid handlers, is traditionally viewed as a drawback.
Existing liquid handling methods have limitations in achieving wide dilution ranges and small volumes.

Purpose of the Study:

To present a novel device and technique that repurposes dispersion for beneficial dilutions.
To demonstrate the Gradient Diluter's capability to extend serial dilution ranges significantly.

Main Methods:

Development of the Gradient Diluter device and associated dispersion techniques.
Validation using fluorescence detection of sulforhodamine and HPLC/UV detection of furosemide.
Application in a T-cell inhibition assay to assess IC50 curve generation.

Main Results:

The Gradient Diluter extends practical serial dilutions over six orders of magnitude.
Achieved precise dilutions in final volumes as low as 10 μL.
Demonstrated comparable IC50 curves to manual methods in a biological assay.

Conclusions:

The Gradient Diluter effectively utilizes dispersion to enhance dilution capabilities.
This novel approach offers improved precision and range for serial dilutions in various applications.
The device shows promise for applications requiring accurate concentration gradients and IC50 determinations.