Palmtop spectrophotometer for DNA and protein measurement in micro-nanoliter assays

Abstract

Spectrophotometer, an important tool in life science, medicine, and analytical fields, usually uses an optical path of 10 mm or more for absorbance measurement of UV light. This corresponds to a sample consumption of ≥ 50 μL in volume and a narrow measuring range of 0.5-50 ng/μL for nucleic acid samples and 0.05-2 mg/mL for protein samples. Higher concentrations must be diluted for measurement. In this paper, we developed an advanced palmtop spectrophotometer for the measurement of both DNA and protein concentrations in micro-nanoliter assays. We constructed a fiber transmission and a fiber reflection absorbance detection scheme illuminated by either UV-LED or deuterium lamp. The sensitivity of 0.5 ng/μL and a wide measuring range of 0.5-2000 ng/μL in concentrations were obtained for DNA, and the sensitivity of 0.05 mg/mL and a wide measuring range of 0.05-100 mg/mL were also obtained for protein. However, sample consumption is only 1 μL in volume for fiber transmission detection scheme and 500 nL for fiber reflection detection scheme. The linear correlation coefficient of measured concentrations to theoretical concentrations is greater than 0.99. With the profit of this work, a miniaturized spectrophotometer with better sensitivity and wider measuring range can be produced for analytical applications.