|On chip micro-extraction and real-time PCR with integrated SPAD optical fluorescence detection for nucleic acid analysis|
Cristina Potrich, Elisa Morganti, Nicola Massari, Lucio Pancher, C. Kostoulas, Laura Pasquardini, Cristian Collini, Andrea Adami, Lorenzo Lunelli, F. Kalatzis, David Stoppa, Cecilia Pederzolli, Leandro Lorenzelli
A PDMS lab-on-a-chip for one step DNA isolation and real time-polymerase chain reaction (RT-PCR) has been designed, fabricated, and characterized for point-of-care clinical diagnostics. In addition, a module for on-chip optical detection based on SPAD - Single-Photon Avalanche Diode - detector has also been developed and used to monitor the presence of specific DNA polymorphisms possibly related to genetic diseases.
|Microfluidic chips with nanostructures for investigation of biological objects by methods of high resolution microscopy|
I.V. KUKHTEVICH 1; A.A. EVSTRAPOV 1,2,3; A.S. BUKATIN 2,3; I.S. MUKHIN 1,2
We have designed and fabricated a microfluidic chips (MFC) with integrated net of nanochannels (traps) for fixation of biological samples in their native environment during study by SPM and CLSM. These traps were created by method of focused ion beam lithography. The width of fabricated nanochannels is in the range from 50 to 300 nm. Fabricated MFC were investigated on test samples in liquid buffer solution.
|Mixed Self-Assembled Monolayers on Bi-functional Magnetic Microcarriers|
D.M. Love, J.J. Palfreyman, K. Vyas, T. Mitrelias, C.H.W. Barnes
We present a bi-functional magnetic microcarrier design, intended for bioassay applications. The chemical functionalisation on the gold side of the microcarriers is optimised using mixed self-assembled monolayers. In-flow quartz crystal microbalance measurements of the formation kinetics are discussed. Fluorescence results are presented.
|Low-volume on-chip single sperm cell analysis|
Pickrahn I. E. ,Schmidt-Gann G., Kroneis T.
We performed low-volume on-chip DNA typing of single sperm cells isolated by means of laser microdissection. 16plex PCR was successful in 39 of 40 single cell samples yielding a mean PCR efficiency of 62.8%. In addition, we were able to identify a single-cell sample containing more than one cell enabling us to monitor the quality of the whole procedure, and hence, exclude “contaminated” samples from further analysis.
|Sequence-independent Selective Amplification of mRNAs over rRNAs|
John Arrand1, Sim Sihota1, Wenbin Wei1 and Guido Krupp2
Standard mRNA amplifications for "All-Exon" microarrays and for bacterial RNAs are impossible with small samples and with degraded RNAs, because removal of rRNAs must precede universal, non-selective RNA amplification.
This pre-treatment with magnetic beads is cumbersome, requires high amounts of starting material, is not universal for all species and degraded RNAs are not suitable.
|Novel Concept of Microarray Construction and their Application in Biology |
B. Cegielska, M. K. Chmielewski, W.T. Markiewicz, M. Figlerowicz
For microarrays produced using spotting technologies, much attention has to be given to the development of slide surfaces, attachment chemistries, and spotting solutions. Application of the optimal and reliable methods ensuring effective binding of DNA probes with slide surface is one of the key factors warranting high quality results. Developments in the field of microarrays occur at a rapid pace and some novel approaches may offer suggestions of new strategies.
|Sub-classification of Colorectal Cancer Using Surface Antigen Antibody Microarray and Fluorescence Multiplexing|
Jerry Zhou, Larissa Belov, Pauline Huang, Joo-Shik Shin, Michael J. Solomon, Pierre Chapuis, Les Bokey, Charles Chan and Richard I. Christopherson
Colorectal cancer (CRC) is the second most frequent cause of cancer deaths in Australia. Even after resection up to 50% of patients relapse. In an attempt to prevent recurrences chemotherapy is administered to high risk patients. However, as few as 10-20% patients genuinely benefit because the clinical course for individuals with CRC remains difficult to predict, largely due to prognostically heterogeneous groups within same-stage tumour categories.
|Hepatitis B virus (HBV) and Human immunodeficiency virus (HIV) antibodies detected by peptide microarrays|
ahmed Abd El Wahed1, Ulrike Beutling2, Ronald Frank2, Gerhard Hunsmann1, and Hans-Joachim Fritz3
HBV and HIVenv chips with overlapping oligopeptides encompassing the full amino acid sequences of HBV and HIV polypeptides were produced. In addition, a chip displaying a library of random 4608 different 15-mers peptides (4608-RPL) was prepared. Both chips were used for analyzing monoclonal antibodies and sera from HIV- and HBV-infected individuals. 4608-RPL could be used for identifying target sequences of antibodies without prior knowledge of the corresponding immunizing antigen.
|System-level Simulation of Liquid Filling in Microfluidic Chips|
Hongjun Song, Yi Wang, and Kapil Pant
The overall objective of our work is to develop a system-level model and simulation framework for investigating the liquid filling process (including the filling time, filling pattern/status, flow velocity/pressure etc.) in complex microfluidic networks with order-of-magnitude speedup over the high-fidelity simulations and without appreciably compromising analysis accuracy.