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Photon-counting microcolorimeter with 40-nl cuvette.

¹Division of Nephrology, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.

The American Journal of Physiology

|July 1, 1991

Summary

A novel 40-nanoliter (nL) cuvette colorimeter was developed, significantly reducing plasma volume needs for the Lowry microprotein assay to 60 picoliters (pL). This microcolorimeter demonstrates linearity and reproducibility for precise protein quantification.

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Area of Science:

Analytical Chemistry
Biochemistry
Instrumentation Engineering

Background:

Traditional microprotein assays require substantial plasma volumes, limiting their application in precious sample analysis.
Existing colorimetric methods often lack the sensitivity and precision for ultra-low volume biological samples.

Purpose of the Study:

To develop and validate a microcolorimeter with a 40-nanoliter (nL) cuvette for ultra-low volume protein quantification.
To assess the performance of the microcolorimeter with the Lowry microprotein assay, focusing on reduced sample volume requirements.

Main Methods:

Construction of a 40-nL cuvette using capillary glass with a microforge-drawn tip and fiber-optic filaments.
Integration of a microscope photon-counting detection assembly for signal measurement.
Application of the microcolorimeter to the Lowry microprotein assay, comparing results with standard methods.

Main Results:

The constructed colorimeter utilizes a 40-nL cuvette, enabling a reduction in plasma volume requirements for the Lowry assay from nanoliters to 60 picoliters (pL).
Verified linearity and reproducibility of the microcolorimeter when used with the Lowry assay.
Colorimeter output was several orders of magnitude above the photon counter's lower limit of detection, indicating high sensitivity.

Conclusions:

The developed 40-nL cuvette microcolorimeter is a sensitive and reproducible instrument for ultra-low volume protein analysis.
This technology significantly minimizes sample volume requirements for established assays like the Lowry method.
The microcolorimeter offers a promising tool for applications involving limited biological samples.