A capillary holder for scanning detection of capillary isoelectric focusing with laser-induced fluorescence.
A holder for a 12 cm long capillary was designed for scanning LIF detection of CIEF. The polyimide coat of a fused-silica capillary has been removed, and 1.5 mm diameter flanges have been attached near both ends. The holder is fixed on the stage of a fluorescence microscope via a translational stage, and a capillary guide is directly fixed on the microscope stage. The guide has a groove and a pressure plate for the capillary to slide in. The holder has two pulling plates with slits of 1 mm to accept the capillary just inside the flanges. The slits and the groove of the guide have been aligned. The motion of the translational stage brings the pulling plate into contact with the flange at the pulled side, and slides the capillary through the guide. The other end of the capillary is free and produces no strain on the capillary. When the motion of the stage is reversed, an unstrained contact is achieved at the other end. The baseline noise from scanning was only 50% larger than that without scanning. The fluorescence-signal variation during scanning was about 4% of the total signal, which was about twice that without scanning.
Takahashi
Katsuyoshi
K
Bioanalytical Instrumentation Laboratory, Department of Applied Chemistry, School of Engineering, The University of Tokyo, Japan.
Maruo
Yuji
Y
Kitamori
Takehiko
T
Shimura
Kiyohito
K
D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis*; D007525:Isoelectric Focusing / Q000295:instrumentation* / Q000379:methods*; D008853:Microscopy / Q000295:instrumentation* / Q000379:methods*
D005456:Fluorescent Dyes