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1.
Biotechniques ; 70(5): 251-262, 2021 05.
Article in English | MEDLINE | ID: mdl-33956504

ABSTRACT

Substantial improvement of microvolume UV absorption spectrometry in sensitivity, robustness and ease of operation was achieved for routine biological applications. A unique microtubing-based absorption cell (208 µm internal diameter) featuring enhanced light transmission with a liquid core waveguide technique provided dramatically enhanced absorption sensitivity, proportional to the extended path length (50 mm, from the typical 1 mm), while robust measurement performance was attained by implementation of preventive measures against bubble trapping along the light path. For pBR322 plasmid DNA, absorbance at 260 nm was reliably measurable down to 0.1 ng/µl with repeatability typically 2-3% RSD. The detection limit was 0.03 ng/µl dsDNA, far lower than the current state-of-the-art âˆ¼1 ng/µl. Sample consumption for each measurement was 2.4 µl. Automated operation implemented for the first time in microvolume spectrophotometry facilitated the ease in handling with small-volume biological samples.


Subject(s)
DNA , Spectrum Analysis , DNA/isolation & purification , Spectrophotometry , Ultraviolet Rays
2.
Ultramicroscopy ; 194: 48-56, 2018 11.
Article in English | MEDLINE | ID: mdl-30071373

ABSTRACT

An independent external stage for low noise atomic force microscope has been developed for mid-range movements so that it aids in measurements of critical dimensions through the low-noise atomic force microscope. The maximum travel length of the external four-axes stage is 10 mm. For image scanning of the specific target region, the sample needs to be moved through two steps: coarse positioning with the external stage and fine positioning with PI XY piezo scanner. Prior to the CD measurements, we confirmed that the position errors caused by the external stage and tip stage were negligible through the reproducibility experiments. In this study, custom-designed software stored the initial position of the probe and then moved it precisely to the sample location to be measured. Subsequently, the sidewalls of an improved vertical parallel structure were measured and the repeatability and reproducibility of the CD measurements were estimated using a CDR30-EBD tip. Finally, we confirmed that tip wear could be minimized by measuring TGX1 samples with undercut structures.

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