Analytical Chemistry Chemical Cytometry Quantitates Superoxide

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Table 3. Evolution of the Sizing Results for Specific Regions of an Allelic Ladder Run for Three Different Chips and Five Different Runs a D22S1045 size of allele no. 7 (bp) size of allele no. 8 (bp) chip 1 - 1st load 304.2 307.3 chip 2 - 1st load 305.6 308.2 chip 2 - 2nd load 304.1 307.2 chip 3 - 1st load 304.3 307.2 chip 3 - 2nd load 304.0 307.2 D18S51 size of allele no. 10 (bp) size of allele no. 10.2 (bp) size of allele no. 11 (bp) chip 1 - 1st load 146.1 148.2 150.2 chip 2 - 1st load 147.1 149.2 151.1 chip 2 - 2nd load 146.3 148.3 150.2 chip 3 - 1st load 145.8 147.7 149.7 chip 3 - 2nd load 145.9 147.8 150.0 TH01 size of allele no. 9.3 (bp) size of allele no. 10 (bp) chip 1 - 1st load 100.2 101.2 chip 2 - 1st load 100.3 101.3 chip 2 - 2nd load 100.2 101.1 chip 3 - 1st load 100.1 101.4 chip 3 - 2nd load 100.2 101.2 FGA size of allele size of allele no. 21.2 (bp) no. 22 (bp) size of allele size of allele no. 22.2 (bp) no. 23 (bp) size of allele size of allele no. 23.2 (bp) no. 24 (bp) size of allele size of allele no. 24.2 (bp) no. 25 (bp) size of allele size of allele no. 25.2 (bp) no. 26 (bp) chip 1 - 1st load 176.8 178.8 180.8 182.8 184.8 186.8 188.9 190.8 192.8 194.8 chip 2 - 1st load 177.2 179.3 181.0 183.0 184.2 186.0 188.0 189.8 191.6 193.3 chip 2 - 2nd load 177.0 178.9 180.7 182.7 184.5 186.6 188.8 190.7 192.6 194.7 chip 3 - 1st load 176.8 178.6 180.7 182.7 184.5 186.6 188.8 190.7 192.6 194.7 chip 3 - 2nd load 176.6 178.5 180.6 182.5 184.7 186.6 188.6 190.6 192.7 194.5 a The values represent the calculated size of the designated allele in DNA base pairs relative to the ILS in the same run. The markers are chosen to represent 1 marker per color, cover most of the regions of interest [100-300 bp], and correspond to the most challenging peaks to separate. Figure 6. Micro Capillary Electrophoresis profile from a fully integrated run. DNA was purified and amplified on-cartridge from lysed cells from a buccal swab and transferred automatically via Teflon tubing to the glass microchip. Allele designation was performed using GeneMarker HID 1.76 (SoftGenetics LLC). version of the software is in development which will use the time file provided by the instrument and so align the data correctly on a true time scale. 6998 Analytical Chemistry, Vol. 82, No. 16, August 15, 2010 CONCLUSION We have demonstrated a successful microTAS approach to the delivery of rapid DNA technology suitable for the forensic
field. DNA profiles were delivered from a crude DNA lysate with no manual input. The automated process is simple to use and provided DNA profiles which were concordant with the expected DNA profile of the donor. Our target is to deliver an evidential quality DNA profile from sample to profile in less than 2 h. While we currently have the capability of processing the sample in under 4 h, we believe that there are a number of areas where we can speed up the process. A major time saving (approximately 1.5 h) can be made simply by reducing the PCR cycling parameters from those published here to thermal profiles similar to those described previously. 4–6 Optimizing the cartridge operating process to allow parallel processing of valve firings and pump control will further reduce the processing time and we believe we will soon be able to meet the target time of 2 h for delivery of the genotyped DNA profile. The turn around between samples is approximately 10 min to load the solutions into the single-use plastic cartridge, and 75 min for preparation of the CE chip. This can be carried out while the previous sample is running, provided that a number of CE chips are available, reducing the turn around time to little more than 5 min. The resolution obtained on the µCE module is very close to that of a commercial CE apparatus and the reproducibility of fragment migration was good: With allelic ladders the maximum variation in the positioning of an allele was around 0.2 bp on the TH01 locus, 0.7 bp on the D18S51 locus, 0.4 bp on the FGA locus, and 0.7 bp on the D22S1045 locus. The migration of DNA profiles from human samples does require some improvement if the system is to be used for probative samples as we were unable to successfully type all integrated runs due to a small variation in the apparent migration of fragments. This variation is likely to be minimized by implementation of a new version of the NanoIdentity software which will take the true time file from the instrument, rather than assuming all data points are equidistant in time. The largest variation in the process remains the amount of template DNA input to the PCR reaction, and while this impacts the peak heights observed in the electropherogram, this does not affect the accuracy of the genotyping of a sample. Simple substitution of the multiplex chemistry in the cartridge will allow the production of DNA profiles suitable for loading to any established national DNA database. However, the rapid delivery of a DNA profile, whether at a crime scene, or in a custody suite is only a part of the puzzle. If the full impact of these new technologies is to be gained, the rapid delivery of chemistry must be supported by a capability to submit and compare the DNA profile in near real time. Currently neither the CoDIS database, nor the UK National DNA database have the capability to support rapid chemistry protocols. While the operation of the instrument is straightforward, a good level of training is required before the instrument can be operated but we believe that any individual with a basic scientific education could become competent in the operation of the instrument. Our vision for the future is to have a single fully integrated injection-molded cartridge with reagents preloaded with capability for DNA purification, amplification and µCE-based separation. Such a cartridge, coupled with automatic channel finding and focusing will minimize set up time and make the system usable by an individual with very basic training. ACKNOWLEDGMENT We gratefully acknowledge the technical assistance of Glen McCarty, David Nguyen, and Brett Duane for electronics design and prototyping; Baiju Thomas, Keith Burt, and Matthew Barrett for plastic device fabrication; Dr. Jian Gu and Peter East for microfabrication; and Amol Surve for instrumentation design. We also thank Bruce McCord for the provision of the separation matrix and helpful discussion around microCE and Promega Corporation for the generous gifts of PowerPlex ESI 16 STR kits and ILS 500 CC5. Received for review May 24, 2010. Accepted July 1, 2010. AC101355R Analytical Chemistry, Vol. 82, No. 16, August 15, 2010 6999
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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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Figure 6. Comparison between simula
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educed in the vicinity of double bo
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Figure 2. Normalized product ion ab
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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
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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
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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
Page 235 and 236: allow the use of higher aptamer con
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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 and 246: Since the time for liquid expulsion
Page 247 and 248: Anal. Chem. 2010, 82, 6991-6999 Int
Page 249 and 250: Figure 1. Representation of the Car
Page 251 and 252: Table 2. Capillary Electrophoresis
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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
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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
Page 297 and 298: Figure 6. (A) FL spectra of BSPOTPE
Page 299 and 300: Scheme 1. Proposed Mechanism for Fl
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Page 303 and 304: 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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Analytical Chemistry Chemical Cytometry Quantitates Superoxide

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