Analytical Chemistry Chemical Cytometry Quantitates Superoxide

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teristics of the producer at least twice. (5) The OE is mainly limited by the required time of pulse volume expulsion. (6) The use of elastic pumping tubes can render higher OE when the flow resistance is high. ACKNOWLEDGMENT B.H. was funded by a JAE postdoctoral fellowship from CSIC. The work was supported from Project CTQ2007-64331 funded by the MEC (Spanish Ministry of Education and Science). 6990 Analytical Chemistry, Vol. 82, No. 16, August 15, 2010 SUPPORTING INFORMATION AVAILABLE Photo of the used 8-port relay card and scheme of the experiment for the determination of the back-spring constant. This material is available free of charge via the Internet at http://pubs.acs.org. Received for review May 17, 2010. Accepted June 28, 2010. AC101250H
Anal. Chem. 2010, 82, 6991–6999 Integrated Microfluidic System for Rapid Forensic DNA Analysis: Sample Collection to DNA Profile Andrew J. Hopwood,* ,† Cedric Hurth, ‡ Jianing Yang, ‡ Zhi Cai, ‡ Nina Moran, † John G. Lee-Edghill, † Alan Nordquist, ‡ Ralf Lenigk, ‡ Matthew D. Estes, ‡ John P. Haley, † Colin R. McAlister, † Xiaojia Chen, ‡ Carla Brooks, ‡ Stan Smith, ‡ Keith Elliott, † Pieris Koumi, † Frederic Zenhausern,* ,‡ and Gillian Tully † Research and Development, Forensic Science Service, Trident Court 2960 Solihull Parkway, Birmingham Business Park, Birmingham UK B37 7YN, and Center for Applied NanoBioscience and Medicine, The University of Arizona College of Medicine, 425 N. Fifth Street, Phoenix, Arizona 85004 We demonstrate a conduit for the delivery of a step change in the DNA analysis process: A fully integrated instrument for the analysis of multiplex short tandem repeat DNA profiles from reference buccal samples is described and is suitable for the processing of such samples within a forensic environment such as a police custody suite or booking office. The instrument is loaded with a DNA processing cartridge which incorporates on-board pumps and valves which direct the delivery of sample and reagents to the various reaction chambers to allow DNA purification, amplification of the DNA by PCR, and collection of the amplified product for delivery to an integral CE chip. The fluorescently labeled product is separated using micro capillary electrophoresis with a resolution of 1.2 base pairs (bp) allowing laser induced fluorescencebased detection of the amplified short tandem repeat fragments and subsequent analysis of data to produce a DNA profile which is compatible with the data format of the UK DNA database. The entire process from taking the sample from a suspect, to database compatible DNA profile production can currently be achieved in less than 4 h. By integrating such an instrument and microfluidic cartridge with the forensic process, we believe it will be possible in the near future to process a DNA sample taken from an individual in police custody and compare the profile with the DNA profiles held on a DNA Database in as little as 3 h. DNA analysis in a forensic context relies on the extraction of DNA from a sample; quantification and normalization of the DNA; concurrent PCR-based amplification of a number of specific short tandem repeat loci and separation/detection of the PCR products, usually by capillary electrophoresis (CE); thereby allowing precise sizing of each fragment and accurate typing of the alleles in the DNA profile. This forensic process allows for the routine analysis of DNA samples from controls such as buccal swabs in 24-72 h * To whom correspondence should be addressed. E-mail: andy.hopwood@ fss.pnn.police.uk (A.H.); Frederic.Zenhausern@arizona.edu (F.Z.). † Forensic Science Service. ‡ The University of Arizona College of Medicine. from receipt of the sample. 1 Such processes are laboratory-based and require that the sample, once taken from a suspect in custody is transported to a laboratory for processing. For convenience, samples are usually stored and batched by the police force prior to dispatch to the lab, a process that can take between a few hours to a few weeks. Even with samples that are dispatched to the laboratory immediately, the suspect is highly likely to have been released from custody while the sample is processed. A sample taken from a suspect in the UK has a 2.3% chance of matching with a crime sample held on the National DNA Database (NDNAD) of the UK. 2 Furthermore, evidence suggests that individuals released on police bail have a high incidence of offending while on bail. 3 It would therefore be beneficial to the arresting agency for the individual to remain in custody while the DNA sample is processed and compared against the NDNAD. This would require the availability of both rapid DNA processing and real-time access to the national database. Within the laboratories of the FSS, current process allows for reference samples in urgent cases to be prioritized and, once delivered to the laboratory, these can be processed manually in as little as 8 h. However, this is a labor intensive and therefore relatively expensive process. The implementation of a rapid system whereby a reference sample can be processed within the police custody area would be of value to the law enforcement community: a suspect’s DNA sample could be processed and compared to a database of crime sample DNA profiles while the individual remains in custody, eliminating the need to locate and rearrest an individual in response to a match on the database. Rapid elimination of an individual from an investigation could also be achieved, freeing up resources for the investigation of alternative leads in a criminal case. A number of publications have reported approaches to the rapid analysis of DNA for forensic science and various methods have been described including the acceleration of specific parts (1) Hedman, J.; Albinsson, L.; Ansell, C.; Tapper, H.; Hansson, O.; Holgersson, S.; Ansell, R. Forensic Sci. Int.: Genet. 2008, 2, 184–189. (2) Annual Report of The National DNA Database 2007-2009. http://www. npia.police.uk/en/docs/NDNAD07-09-LR.pdf (Accessed March 2010). (3) Morgan, P. M.; Henderson, P. F. Home Office Report 184 1998 ISBN 1 84082 074 8 http://www.homeoffice.gov.uk/rds/pdfs/hors184.pdf (Accessed Feb. 2010). 10.1021/ac101355r © 2010 American Chemical Society 6991 Analytical Chemistry, Vol. 82, No. 16, August 15, 2010 Published on Web 07/15/2010
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Anal. Chem. 2010, 82, 6745-6750 Let
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purchased from Invitrogen-Molecular
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Figure 3. Electropherograms of TPP-
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Anal. Chem. 2010, 82, 6751-6755 Res
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(pH 8.0) with cysteine and cystamin
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Figure 4. Ion mobility mass spectra
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GOx glucose + O298 gluconic acid +
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coater at 2000 rpm for 20 s, and th
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Figure 5. Comparison of cyclic volt
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in channels with either no grooves
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indicators of atmospheric processin
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Figure 1. GC/MS total ion chromatog
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Table 2. Concentrations and Stable
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on a substrate are preferred. 20-24
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Figure 4. SERS analysis of NAADP co
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Anal. Chem. 2010, 82, 6775-6781 Hig
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tion, 2 µL of proprionaldehyde wer
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Figure 4. Analysis of 2a by HPLC-MS
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eaction of 1a with PBH can be condu
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Numerous references had demonstrate
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Figure 1. TEM images of the prepare
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Figure 4. Schematic representation
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esult in a big SPR signal change wi
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also reduces chemical noise, which
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Table 1. Extraction Yields, Liquid
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scanning of AMPP amides of the anal
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Anal. Chem. 2010, 82, 6797-6806 δ
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Figure 1. Schematic view of the pre
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from the specimen and enclosed in a
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Figure 4. (A) IRMS mass-44 chromato
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Table 3. Ambient Measurement Result
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Anal. Chem. 2010, 82, 6807-6813 Dir
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Polymerase (1 U per sample). Reacti
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of which was constant for all ampli
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analytically useful signals at less
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carbon black and RP-C18 for the ext
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solution in an equal volume, and 1
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Table 2. Concentrations and Ratios
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Anal. Chem. 2010, 82, 6821-6829 Mac
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mg/mL protein, followed by separati
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Figure 2. Productivity of SEQUST an
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Figure 4. High-resolution MS/MS spe
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Figure 6. Characterization of PSMs
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containing T-T mismatches. 23 Based
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Figure 1. Extinction spectra of sol
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Figure 3. (A) The value of Ex650 nm
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Figure 5. Extinction spectra and co
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wished to explore the dehydration o
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constant medium for separation; we
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Figure 2. Standards of (Pi)n, n ) 1
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Figure 5. Quantitative calibration
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Anal. Chem. 2010, 82, 6847-6853 Met
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Figure 1. 226 Ra spectrum by liquid
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Table 1. Counting Properties and De
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Table 3. Analysis of 226 Ra in Sedi
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mercial microarray scanner and fabr
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Figure 2. Optical transmission meas
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Figure 5. Volcano plots detailing t
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data as well. The 41 genes in Table
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corresponding compound if its chemi
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Figure 1. Schematic illustration of
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Table 1. Absolute Quantification Re
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ment. Therefore, the long-time drea
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Figure 1. Chemically actuated micro
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Figure 2. Influence of a surfactant
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Figure 5. Device to eject and mix s
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Anal. Chem. 2010, 82, 6877-6886 Imm
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NaCl, phosphate buffer saline (PBS)
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Figure 2. Product ion mass spectra
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-70 °C resolved the problem, givin
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Table 1. Intraday Precision and Acc
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Anal. Chem. 2010, 82, 6887-6894 Fer
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Figure 1. Infrared spectra of (A) u
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Figure 3. Cyclic voltammograms obta
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Figure 6. Calculated charge from ch
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Anal. Chem. 2010, 82, 6895-6903 Ele
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Table 1. Chemical Structure, pKa Va
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pH with a tilted baseline (Figure 1
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Table 2. Linearity and Detection Li
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ascorbic acid (AA), uric acid (UA),
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the multielement capabilities, the
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RESULTS AND DISCUSSION Sulfur Detec
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Table 2. Molecular Properties and C
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Anal. Chem. 2010, 82, 6911-6918 Dir
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dilution and hybridization buffer.
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solution under appropriate incubati
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Figure 4. Standardization curve for
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Anal. Chem. 2010, 82, 6919-6925 Ele
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the ×10 objective, to have a large
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Figure 2. With a suitable removal o
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the NB signal in a much better foot
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Scheme 1. Reactions of Selenium Rea
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Figure 2. (a) ESI-MS spectrum showi
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Figure 4. (a) ESI-MS spectrum showi
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Anal. Chem. 2010, 82, 6933-6939 Dif
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trode 28 by a finite element using
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Figure 4. Comparison between simula
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Page 197 and 198: educed in the vicinity of double bo
Page 199 and 200: Figure 2. Normalized product ion ab
Page 201 and 202: Figure 4. EID (a) and IRMPD (b) of
Page 203 and 204: Anal. Chem. 2010, 82, 6947-6957 Ide
Page 205 and 206: Figure 1. Schematic flowchart showi
Page 207 and 208: difference, ppm compound Table 1. I
Page 209 and 210: isoforms, its successful use, in th
Page 211 and 212: Figure 5. Extracted ion current ESI
Page 213 and 214: m/z 1172.935 was observed for Ser14
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Page 217 and 218: Scheme 2. Fragmentation Mechanism o
Page 219 and 220: Figure 1. (A) ESI-LTQ-CID-MS 2 prod
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Page 223 and 224: Figure 5. (A) MALDI-TOF/TOF product
Page 225 and 226: Anal. Chem. 2010, 82, 6969-6975 Ana
Page 227 and 228: Figure 3. Equilibrium response as a
Page 229 and 230: Figure 6. The average measured resp
Page 231 and 232: for the fill time, and we find that
Page 233 and 234: nucleotide tails. 3-5 Thus, the amo
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Page 239 and 240: Anal. Chem. 2010, 82, 6983-6990 Imp
Page 241 and 242: Figure 2. Configuration editor wind
Page 243 and 244: Figure 4. Dependencies between volu
Page 245: Since the time for liquid expulsion
Page 249 and 250: Figure 1. Representation of the Car
Page 251 and 252: Table 2. Capillary Electrophoresis
Page 253 and 254: Figure 4. (A) Typical raw electroph
Page 255 and 256: field. DNA profiles were delivered
Page 257 and 258: Another approach to handle this lim
Page 259 and 260: (principal components, PCs) in whic
Page 261 and 262: Figure 5. Relevant score plots and
Page 263 and 264: CONCLUSIONS In this paper we have i
Page 265 and 266: electroactive-species loaded liposo
Page 267 and 268: Figure 2. Qdot-based FLFTS response
Page 269 and 270: Figure 6. Fluorescence imaging of Q
Page 271 and 272: Anal. Chem. 2010, 82, 7015-7020 Sel
Page 273 and 274: Table 1. Effect of 1 D-LC Condition
Page 275 and 276: Figure 3. Peak-production rate vers
Page 277 and 278: Anal. Chem. 2010, 82, 7021-7026 Cel
Page 279 and 280: luciferase (T7 control vector) was
Page 281 and 282: Figure 3. Inhibitory effects of luc
Page 283 and 284: Anal. Chem. 2010, 82, 7027-7034 Pat
Page 285 and 286: Electrochemistry. Electrochemical m
Page 287 and 288: Figure 2. SEM images of Au substrat
Page 289 and 290: Table 3. Film Thickness Measurement
Page 291 and 292: Anal. Chem. 2010, 82, 7035-7043 Qua
Page 293 and 294: Figure 1. (A) FL spectra of BSPOTPE
Page 295 and 296: Figure 4. (A) Variation in the FL i
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Figure 6. (A) FL spectra of BSPOTPE
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Scheme 1. Proposed Mechanism for Fl
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one or more drawbacks including poo
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Figure 2. Free fluorophores do not
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Anal. Chem. 2010, 82, 7049-7052 Dev
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Figure 2. Comparative analysis of d
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