ON-CHIP DROPLET ENHANCED FLUORESCENCE EMISSION
FOR LOW CONCENTRATION PROTEIN MEASUREMENT
Y. F. Yu
1, 2
, T. Bourouina
2
, and A. Q. Liu
1
1
School of Electrical & Electronic Engineering, Nanyang Technological University, Singapore 639798
2
Ecole Supérieure d'Ingénieurs en Electronique et Electrotechnique, University of Paris Est, France, 93162
(Email: eaqliu@ntu.edu.sg , Tel: +65-6790-4336, URL: http://nocweba.ntu.edu.sg/laq_mems/ )
ABSTRACT
On-chip droplet enhanced fluorescence emission for low
concentration protein measurement is presented in this
paper. The system includes T-junction of microchannel for
micro-droplet generation and two integrated optical fibers
for excitation laser input and fluorescence detection. The
excitation light is confined in the droplet by total internal
reflection to increase the optical path length, which
enhances the fluorescence emission. The on-chip droplet
features higher stability and easier manipulation. The size
of the droplet can be varied by changing the flow
conditions. The intensity of the droplet enhanced
fluorescence emission is much higher than the intensity of
the free space fluorescence, which promises wide
applications in biological detection and measurement.
KEYWORDS
Droplet, Microfluidics, Fluorescence emission,
Microphotonicfluidic systems and Optofluidics.
INTRODUCTION
Traditional methods for protein concentration measurement
are mostly based on absorption techniques, such as
ultraviolet absorption, alkaline copper, bicinchoninic acid,
Coomassie blue staining or gold nano- particle labeling [1].
These methods have limitations on detection sensitivity,
quantitative accuracy and compatibility with modern
protein identification and characterization procedures as
compared with the fluorescence based protein detection
methods [2]. In on-chip droplet enhanced fluorescence
emission, the intensity of fluorescence emission is
increased using a spherical cavity [3]. The intensity
enhancement has real significance in increasing the
sensitivity of measurement. Fluorescence intensity is
quantitatively depending on the molar extinction
coefficient, the optical path length and the solute
concentration, the excitation light intensity and the
fluorescence collection efficiency of the system. But for the
same sample solution, the molar extinction coefficient and
the solute concentration are constants. The adjustable
parameters are the optical path length and the fluorescence
collection efficiency. They are directly proportional to the
fluorescence intensity. In order to increase the optical path
length, the excitation light is confined in a spherical cavity
(here is a droplet) by total internal reflection. Due to the
interfacial tension between two immiscible liquids, the
multiphase liquid droplet has ultrahigh smooth interface,
which insures low loss. The optical path length can be
lengthened by thousand times through resonance. In this
paper, a micro-optic-fluidic system is applied to replace the
traditional electrophoresis detection method [4]. Optical
fibers integrated in the chip for spectrum detection
increases the fluorescence collection efficiency because the
fibers are close to the droplets [5]. The micro-optic-fluidic
chip is suitable for experiments with tiny volume of sample,
which reduces the total cost and accelerates the detection
time. Compared to the aerosol droplets for detection [6], the
on-chip droplets have higher stability and simpler
manipulation.
DESIGN AND ANALYSIS
The micro-optic-fluidic chip consists of two liquid inlets
and a waste outlet, a two-inlet t-junction for multiphase
plugs generation, an enlarged channel region and two
integrated optical fibers as shown in fig. 1. The plugs are
reformed into droplets due to the surface tension, and the
length of the plugs determines the diameter of the droplets,
which is determined by the flow conditions.
For optical measurement, one of the methods is to input the
excitation light from the top and use a CCD device to
capture the images with a filter. Alternatively, the excitation
light is injected by the optical fiber and the emission is
collected by another optical fiber and detected by the
Figure 1: Schematic of (a) optofluidic chip with optical
detection (b) detection region.
Excitation laser
Fluorescence detection
Oil inlet
Glycerin inlet
Outlet
(a)
(b)
978-1-4244-4193-8/09/$25.00 ©2009 IEEE Transducers 2009, Denver, CO, USA, June 21-25, 2009 1250
T3P.093