Heterogeneous Integration of a LASER Induced Fluorescence Detection Device for Point-of-Care Microfluidic Biochemical Analysis

2012 ◽  
Vol 1426 ◽  
pp. 211-222
Author(s):  
Toshihiro Kamei

ABSTRACTFluorescence detection is more advantageous than electrochemical detection in terms of high sensitivity, multiplexed detection capability and isolation from analyte. Integration of fluorescence detection, however, is much more difficult. First, it would require heterogeneous integration of various optical components including an excitation source, an optical filter, a lens, a mirror and a detector. Second, most of integrated fluorescence detectors, even though not fully integrated, suffer from high limit of detection (LOD) compared to conventional optical system that consists of discrete optical components. We have reduced laser light scattering in an integrated hydrogenated amorphous Si (a-Si:H) fluorescence detector, significantly improving a limit-of-detection (LOD). The detection platform comprises a microlens and the annular fluorescence detector where a thick SiO2/Ta2O5 multilayer optical interference filter is monolithically integrated on an a-Si:H pin photodiode. With a microfluidic capillary electrophoresis (CE) device mounted on the platform, the integrated system is demonstrated to separate DNA restriction fragment digests with high speed, high sensitivity and high separation efficiency, implying single molecular DNA detection when combined with polymerase chain reaction (PCR). We are now working towards integration of an excitation source to fabricate heterogeneously integrated laser-induced fluorescence detection (LIF) device that would be comprised of an InGaN laser diode, microlenses and the integrated a-Si:H fluorescence detector.

2007 ◽  
Vol 61 (7) ◽  
pp. 777-779 ◽  
Author(s):  
Kimia Sobhani ◽  
David A. Michels ◽  
Norman J. Dovichi

The sheath-flow cuvette is a key component in a high-sensitivity post-column laser-induced fluorescence detector for capillary electrophoresis. Most designs are based on commercial cuvettes originally manufactured for use in a flow cytometer. In these devices, a quartz flow chamber is held in a stainless-steel fixture that is difficult to machine and subjects the cuvette to a torque when sealed, which frequently leads to damage of the flow chamber. In this report we present a design for a cuvette that may easily be constructed. This design uses compression to hold and seal the quartz flow chamber without applying torque. The system produces detection limits (3σ) of 115 yoctomoles (70 copies) for FQ-labeled carbonic anhydrase.


1991 ◽  
Vol 37 (9) ◽  
pp. 1492-1496 ◽  
Author(s):  
J Z Zhang ◽  
D Y Chen ◽  
S Wu ◽  
H R Harke ◽  
N J Dovichi

Abstract Capillary electrophoresis provides very high efficiency separations of biological molecules. Laser-induced fluorescence produces very high sensitivity detection. The combination of the two techniques results in an analytical tool with unprecedented properties: separations with more than a million theoretical plates and detection limits of a few hundred analyte molecules. This paper considers the design of high-sensitivity laser-induced fluorescence detection for capillary zone electrophoresis separation of labeled amino acids and capillary gel electrophoresis separation of DNA sequencing fragments.


2020 ◽  
Vol 92 (13) ◽  
pp. 8680-8684 ◽  
Author(s):  
Qiuying Xi ◽  
Meng Shi ◽  
Xuhui Geng ◽  
Xiaona Wang ◽  
Yafeng Guan

1999 ◽  
Vol 45 (6) ◽  
pp. 862-868 ◽  
Author(s):  
Steinar Hustad ◽  
Per Magne Ueland ◽  
Jørn Schneede

Abstract Background: Riboflavin is the precursor of flavin mononucleotide (FMN) and FAD, which serve as cofactors for several redox enzymes. We have developed a capillary electrophoresis method for the determination of riboflavin and its two coenzyme forms in human plasma. Methods: Trichloroacetic acid-treated plasma was subjected to solid-phase extraction on reversed-phase columns. The analytes were separated by micellar electrokinetic capillary chromatography in uncoated fused- silica capillaries filled with borate buffer containing 50 mmol/L sodium dodecyl sulfate, methanol, and N-methylformamide. Native fluorescence was monitored at 530 nm, using an argon laser operating at 488 nm as excitation source. Results: The assay was linear over a concentration range of two orders of magnitude, and the limit of detection was far below physiological concentrations for all vitamers. The within-day and between-day coefficients of variation were 4–9% and 6–12%, respectively. The reference values (median, 5–95 percentiles) obtained by analyzing plasma from 63 healthy subjects were 8.6 nmol/L (2.7–42.5 nmol/L) for riboflavin, 7.0 nmol/L (3.5–13.3 nmol/L) for FMN, and 57.9 nmol/L (44.5–78.1 nmol/L) for FAD. Conclusions: Capillary electrophoresis with laser-induced fluorescence detection allows determination of all riboflavin vitamers far below physiological concentrations. The method may become a useful tool for the assessment of riboflavin status in humans.


2014 ◽  
Vol 53 (6S) ◽  
pp. 06JL02 ◽  
Author(s):  
Toshihiro Kamei ◽  
Keiko Sumitomo ◽  
Sachiko Ito ◽  
Ryo Takigawa ◽  
Noriyuki Tsujimura ◽  
...  

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