RESEARCH ARTICLE
Copyright © 2012 American Scientific Publishers
All rights reserved
Printed in the United States of America
Journal of
Biomedical Nanotechnology
Vol. 8, 1–6, 2012
Development of a Total Temperature Micro-Volume
Blended Incubating and Hybridizing Apparatus for
DNA Hybridization on Nanoparticles
Bin Liu
1†
, Zhiyang Li
1 2†
, Hui Chen
1
, Yan Deng
1
, Nongyue He
1 ∗
,
Sauli Elingarami
1 3
, and Jin Huang
4
1
State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China
2
College of Life Science, Yangtze University, Jingzhou 434025, China
3
Ministry of Health and Social Welfare, Department of Preventive Services, TFNC, Ocean Road 22,
P. O. Box 977, Dares Salaam, Tanzania
4
Nanjing Longliang Biological Science and Technology Limited Company, 210000, China
With the wide application of nanomaterials in biomedical detection in recent years, hybridization
methods which use nanoparticles as solid phase hybridization carriers have emerged. However,
commercial equipments, such as conventional thermal cyclers and hybridization ovens are usually
not appropriate for DNA hybridization on the surface of nanoparticles. We designed and improved
a total temperature micro-volume blended incubating and hybridizing apparatus (TTMHA), which
can be used for blending and suspending of nanoparticles in a small volume and liquid phase
environment. This device highlights the mechanical rotation structure which can not only provide a
uniform temperature field, but also makes the liquid flow fully in the reaction system and improves
DNA hybridization efficiency significantly. A complex PID control algorithm, including Bang–Bang
control and Fuzzy-PID control algorithm, was applied in this research project to improve the con-
trol accuracy and stability. Furthermore, a model detection experiment using ssDNA (single strand
DNA) sequence was conducted in thermal cyclers and TTMHA respectively to verify the optimal
hybridization efficiency of the TTMHA.
Keywords: DNA Hybridization Apparatus, Nanoparticles, Fuzzy PID Control Algorithm.
1. INTRODUCTION
Because of size effect, quantum size effect, surface effect
and macro-quantum tunnel effect, nanoparticles have been
becoming the largest and most widely used nanomaterials
in biomedical engineering and clinical diagnosis research
today.
1–5
They are usually applied in pathogen detec-
tion, early diagnosis of tumor markers, visual detection of
diseases in vitro and other aspects.
6–12
With the wide application of nanomaterials in biomed-
ical detection in recent years, the hybridization method
which uses nanoparticles as solid phase hybridization car-
riers is becoming the main focus today. For biomedi-
cal application, the surface of nanoparticles and other
nanomaterials should be modified with chemical and bio-
logical macromolecules such as proteins, carbohydrates
and oligonucleotide probes.
13 14
Particularly, the specific
∗
Author to whom correspondence should be addressed.
†
These two authors contributed equally to this work.
binding, such as antibodies and antigens, nucleic acid
hybridization, on the surface of nanoparticles requires
a certain temperature which is usually a crucial factor.
Therefore, the nanoparticles hybridization should be put in
a machine with precise temperature. As listed in Table I,
there are mainly three kinds of hybridization apparatuses
on the market as classified by applicable solid phase
materials. Through a variable angle rotating platform, the
hybridization apparatus, based on glass slide or micro-
titer plate, can make the inside liquid flow and mix the
hybridization solution. Membrane hybridization has been a
slow process carried out on a plane and usually put in the
50 mL tubes because it needs more hybridization solution.
In these commercial hybridization apparatuses, the motor
shaft is usually paralleled to the tubes which drive them
by scrolling or rotating horizontally and the hybridization
solution is continuously mixed with its own gravity action.
However, the required liquid volume for nanoparticles
hybridization usually ranges from 20 L to 100 L.
Because of capillary action, the hybridization solution can
J. Biomed. Nanotechnol. 2012, Vol. 8, No. 6 1550-7033/2012/8/001/006 doi:10.1166/jbn.2012.1453 1