ライフサイエンスコーパス: pipette

1 (volumetric flasks, graduated cylinders, and pipettes).

2 by infusion of GluA4 antibody through patch pipette.

3 ear spiral, up to 180 mum from the recording pipette.

4 a(2+) is chelated by 10 mM EGTA in the patch pipette.

5 nts upon inclusion of Kir2.1-Ab in the patch pipette.

6 rnally and intracellularly through the patch pipette.

7 cell-attached patches through the recording pipette.

8 nts with trypsin added to the outside of the pipette.

9 5,10-imine maleate was included in the patch pipette.

10 earance of large vesicles around the puffing pipette.

11 f FXYD1 KO and WT myocytes through the patch pipette.

12 cinity of the dendritic whole-cell recording pipette.

13 loaded with the NO probe DAF-FM via a patch pipette.

14 phosphoinositides infused through the patch pipette.

15 oduced into one cell of a pair using a patch pipette.

16 recorded extracellularly with a loose-patch pipette.

17 GOmega) continuously pulls membrane into the pipette.

18 injection into the neuron through the patch pipette.

19 o automated sample handlers and multichannel pipettes.

20 n 5-10min following cell break-in with patch pipettes.

21 ptimized microfluidic device integrated with pipettes.

22 e used to control injections from very small pipettes.

23 with 200 muL of 0.9% NaCl buffer directly by pipetting.

24 , followed by rinsing, we were able to reuse pipettes 10 times with no degradation in signal fidelity

25 g SLT-1B from complex cell lysates simply by pipetting 20 muL of the sample in and out of the tip in

26 s down to approximately 100 nL, although the pipetting accuracy and precision deteriorate considerabl

27 le trace amounts of Alconox remaining in the pipette after cleaning did not affect ion channel pharma

28 s practice of manually replacing patch-clamp pipettes after each recording.

29 blocker, 4-aminopyridine (4-AP, 5 mM) in the pipette also antagonised the effects of U6.

30 igh-throughput format using an eight-channel pipette and a homemade mini-heater, with a maximum throu

31 reagents are delivered using a multichannel pipette and a microwave reactor is used to complete pept

32 usion of 100 microM diC8-PIP(2) in the patch pipette and bathing solutions, respectively, inhibited a

33 on of Kv2.1 antibodies through the recording pipette and extracellular application of rStromatoxin-1

34 First, unless the patch pipette and GUV pressures are precisely matched in the G

35 ucing the tip size, the ability to reuse the pipette and ion exchange with the cytoplasm.

36 rious [ATP], [InsP3] and [Ca2+] in the patch pipette and measured single InsP3R channel activity in s

37 blocked by phosphatase delivered within the pipette and not affected by treatment with the phosphata

38 ntents are aspirated through the patch-clamp pipette and prepared for RNA-sequencing.

39 iI were placed onto the tips of pulled glass pipettes and inserted into the inner nuclear layer of fi

40 en times smaller than typical microinjection pipettes and rather than pressure pulses as delivery met

41 table clamp, so on-chip operation only needs pipetting and adjusting of clamping force.

42 ple microwells thus avoiding repeated manual pipetting and costly robots.

43 Pipetting and dilution are universal processes used in c

44 ocular-surface epithelia can be isolated by pipetting and FACS sorting into a population of corneal

45 abricated using both manual liquid handling (pipette) and an automated liquid dispensing platform, th

46 n potentials were recorded with a whole-cell pipette, and the corresponding axonal signals were recor

47 ell in focus while iteratively adjusting the pipette approach trajectory to compensate for tissue mot

48 n to compensate for cell movement as a patch pipette approaches a targeted neuron.

49 ly, when membranes contain Ni(2+) complexes, pipetting aqueous polyhistidine-tagged protein through t

50 Existing pipettes are capable of delivering fluids with attolitre

51 By using a carbon pipette as the tip in the scanning electrochemical micro

52 gly curved cylindrical (tube) membrane and a pipette-aspirated giant unilamellar vesicle.

53 P2 hydrolysis, blunted by PIP2 in whole-cell pipettes, attenuated by expression of PIP2-sequestering

54 lecting duct principal cell line using patch pipettes back-filled with a solution containing phosphat

55 allow as the approach angle of a patch-clamp pipette between a water immersion objective and the spec

56 elivered into rods and cones through a patch pipette, binds to and dissociates from synaptic ribbons.

57 mixing, the sample was transferred from the pipette bulb to a small diameter tube, and captured biom

58 included these two nucleotides in the patch pipette but found that TRPC5 currents were absent or wer

59 d for the two cell preparations at the lower pipette Ca2+ levels.

60 Whole cell K+ currents were determined at pipette [Ca2+] of 100 nM or 5 microM in the presence and

61 (length, 50-60 microm) at the tip of a glass pipette, can probe the intracellular environment with a

62 ABA(B) receptors blocked and Cs+ as the main pipette cation.

63 Inclusion of PKA in the pipette caused a negative shift in the voltage dependenc

64 sed firing rate linked to an increase in the pipette-cell conductance.

65 We demonstrate the utility of pipette cleaning by developing the first robot to perfor

66 when applied onto plants in droplets with a pipette compared with a fine spray inoculation.

67 was abolished by GDP-betaS in the recording pipette, consistent with a G-protein-activated inwardly

68 anger blocker KB-R7943, or with BAPTA in the pipette, consistent with a mechanism based on activation

69 mple was mixed within the bulb of a transfer pipette containing lyophilized nucleic acid-silica adsor

70 preconcentration, and clean-up method using pipettes containing immobilized metal ion affinity chrom

71 llular InsP3 via membrane rupture by a patch pipette (containing InsP3)in myocytes expressing FIRE-1

72 ameterized the model with respect to suction pipette current recordings from single cells stimulated

73 amma subunits were included in the recording pipette, D1 but not D2 agonists depolarized layer I neur

74 A simple pipetting device was developed for reliable application

75 ic depression and LTP inhibition after patch-pipette dialysis.

76 Intracellular delivery of GA via a patch pipette did not potentiate the acute effect of GA on Kir

77 oading neurons with GDP-beta-S via the patch pipette did not reverse Rem2-mediated calcium channel in

78 ed or free fluorophores via whole-cell patch pipette did not support photopotentiation.

79 solution (<2 muL) and the MNPs (1-3 mug) by pipetting directly onto a matrix-assisted laser desorpti

80 ations by directly drawing micropillars from pipette-dispensed PDMS microdroplets using vacuum-chucke

81 nstant, increased fluid flow from a separate pipette during current recording.

82 vectors could be delivered through the patch-pipette during such recordings and that these vectors dr

83 Pipette effects on cell firing are traced to a combinati

84 obtained by rigidly fixing ('anchoring') the pipette electrode to the head; however, previous anchori

85 ole cell transduction currents through patch pipette electrodes at the basolateral membrane.

86 ast, and automated method for cleaning glass pipette electrodes that enables their reuse within one m

87 o the base of the outer segment (outside the pipette) elicited a slow inverted response.

88 (3 microM) into the cell with the whole-cell pipette enabled AMPH-induced DA efflux at -60 mV in both

89 r infusion of autoactivated CaMKII via patch pipette enhanced chloride currents 3-fold, and this regu

90 r step causing inaccurate pipetting or total pipetting failure.

91 Inclusion in the pipette-filling solution of a peptide corresponding to t

92 n of the Ca2+-specific chelator BAPTA in the pipette-filling solution or preincubation with the calci

93 Applying moderate suction to the pipette flattens the membrane, reducing tension, and mak

94 and transferred into lysate buffer by mouth pipette, followed by reverse transcription carried out d

95 range fluidic/electrical connections so that pipetting functions can be performed conveniently.

96 ative pressure (suction) applied through the pipette had no effect on the channel, but positive press

97 Pipette-held test objects are translated perpendicularly

98 A and DAG lipase inhibitors in the recording pipette; however, unlike in PCs, NMDA receptors (NMDARs)

99 hole-cell recordings with Neurobiotin-filled-pipettes in horizontal slices from adult rat spinal cord

100 technique that rapidly (within 15 s) anchors pipettes in place with virtually no movement, thus subst

101 dicator fluo-4 in the whole cell patch clamp pipette, in addition to 20 mM EGTA and other constituent

102 ter mix, and detection reagents with minimal pipetting, in a hand-held, disposable device intended fo

103 With zero Na+ in the pipette INa inactivation produced a decline in the SR Ca

104 Pipette inclusion of protein kinase A (PKA) catalytic su

105 catalase, a peroxidase enzyme, in the patch pipette increased the spontaneous firing rate of all dop

106 (L-GSH), through the intracellular recording pipette, indicating that the increased NMDAR response wa

107 ted effect on EGF-induced conductance, while pipette infusion of phosphatidylinositol-3,4,5-trisphosp

108 Pipette infusion of purified phosphatidylinositol-4,5-bi

109 ysis of DiC(8) PI(4,5)P(2) through the patch pipette inhibited Ca(2+)-dependent inactivation of TRPV6

110 tdIns(4,5)P2 or PtdIns(4)P through the patch pipette inhibited desensitization of TRPV1, indicating t

111 on of a small competing peptide in the patch pipette inhibited the SKF 81297-induced reduction in pea

112 he phosphoinositide 3-kinase pathway, in the pipette inhibited these effects.

113 e or saline or the insertion of the infusion pipette into the SI cortex of the right hemisphere.

114 m chloride, 1 mL of acetonitrile extract was pipetted into a 2-mL centrifuge tube containing anhydrou

115 aliquots of batch-cultured cells are rapidly pipetted into a hypotonic medium.

116 Samples are pipetted into an array of separable, multiplexed affinit

117 -attached mode, on pipette potential, and on pipette ionic composition.

118 A multi-pipette is the only additional equipment required for hi

119 The tip size of these pipettes is approximately ten times smaller than typical

120 nometres that resides at the apex of a sharp pipette: it provides scanning cryogenic thermal sensing

121 racellular Ni(2+), inclusion of BAPTA in the pipette, KB-R7943, and SKF96365.

122 In separate pipette-loading experiments, the intracellular diffusion

123 Samples are not pipetted manually but deposited by dragging one or sever

124 sily completed by either manual or automated pipetting methods.

125 the laboratory include traditional transfer pipettes, micropipettes based on air displacement, and m

126 ment by tracking cell position and adjusting pipette motion while approaching a target.

127 of a double-barrel (theta-glass) application pipette mounted on a piezo actuator.

128 dures were slow and often caused substantial pipette movement, resulting in loss of the recording or

129 to the above values, despite changes in the pipette Na(+) concentration, showing autoregulation of N

130 oss numerous 96 well plates and guidance for pipetting, non-trivial tasks for which informatics and v

131 High concentrations of PIP(2) in the patch pipette not only resulted in a strong increase in sperm

132 3 and by intracellular dialysis from a patch-pipette of activated (thiophosphorylated) recombinant AM

133 influx was prevented by the presence in the pipette of the extracellular chaperone clusterin, which

134 oyl phosphatidylcholine (bis-DenPC) on glass pipettes of approximately 10 microm (I.D.).

135 ereafter "chicken matrix"), with just single pipetting of sample (i.e., no filtration, culturing and/

136 sample processing is required apart from the pipetting of the sample into a tin foil cup, which is pl

137 after saline infusion and by T = 3 h in the pipette only group.

138 e-injected group and T = 4 h in the infusion pipette only group.

139 manually dragging this liquid crystal from a pipette onto salty media, it is possible to extend this

140 EMD ranging from 15 to 125 ng/5 microl were pipetted onto a 3-mm diameter x 2-mm thick bioabsorbable

141 For loading, the suspended microcrystals are pipetted onto the chip and excess mother liquor is subse

142 we found that pungent chemicals added to the pipette or bath solution easily activated TRPA1 in cell-

143 r through direct Na(+) loading via the patch pipette or by focal application of glutamate (20 mM puff

144 sponse to cytosolic NADH loaded directly via pipette or increased by extracellular lactate perfusion,

145 many areas of science and technology, where pipettes or related devices are used for dispensing well

146 ated sample-transfer step causing inaccurate pipetting or total pipetting failure.

147 ith the spatial resolution controlled by the pipette orifice radius and a few nanometers film thickne

148 y the nanoparticle translocation through the pipette orifice.

149 Patches creep up the pipette over time with voltage independent and voltage d

150 channels, local [Ca(2+)] imaging, and patch pipette perfusion of EGTA at the calyx of Held.

151 Cplx3 was introduced via a whole-cell patch pipette placed directly on the synaptic terminal, and ve

152 Adaptive pipette positioning provides a platform for future advan

153 Making the pipette potential more positive leads to an increase in

154 te depends on time in cell-attached mode, on pipette potential, and on pipette ionic composition.

155 reversed the direction of creep at positive pipette potentials.

156 itutively active PKC delivered via the patch pipette potentiated NMDA (but not AMPA) whole-cell curre

157 eurons performed with high- and low-chloride pipettes (predicted GABA(A) receptor reversal potentials

158 uter segment drawn part way into the suction pipette, presentation of flashes to the base of the oute

159 The axial deformation of a pipette-pressurized fluid membrane bag produces minuscul

160 clusion of GDP-betaS (1 mM) in the recording pipette prevented all of the U6 effects, suggesting that

161 itor peptide (PKCepsilon-I) in the recording pipette prevented the reduction of peak Na+ current by O

162 carboxylicacid hexyl ester] in the recording pipettes prevented modulation.

163 evice without loss of tumor cells during the pipetting process.

164 reshly prepared reagents within the transfer pipette produced similar results, suggesting that long-t

165 ultaneous whole-cell patch-clamp and suction pipette recording revealed that both the fast and slow c

166 eflects rapid exit through membrane near the pipette recording site.

167 ersed rabbit portal vein myocytes with patch pipette recording techniques.

168 Here we used suction-pipette recordings from isolated ORNs of OMP(-/-) mice t

169 ammalian cells compare well with traditional pipette recordings.

170 ell level as observed by single-cell suction pipette recordings.

171 eurons with natural Cl- concentration (patch pipette removed).

172 Media and drugs are supplied by a pipetting robot system.

173 To close this gap, we developed Lego-based pipetting robots that reliably handle liquid volumes fro

174 low-cost household consumables, programming pipetting routines, and modifying robot designs, we enab

175 -1-induced and direct InsP3 application (via pipette rupture) revealed that InsP3 diffusion into the

176 Low pH pipette saline also increased E(a) and reduced the seal

177 High ionic strength pipette saline increased E(a) and, surprisingly, increas

178 hannel blocker MK-801 was added to the patch-pipette saline, suggesting that postsynaptically express

179 herichia coli and Salmonella within a single-pipetted sample.

180 Great advancements in harvest (in vivo pipette), sample preparation, and sequencing (Illumina H

181 of voltage between pairs of perforated-patch pipettes sealed onto abluminal cells located on microvas

182 With manual pipetting, setting up a plate takes about 2 h, the initi

183 Imaging with fluorescent dyes in the pipette showed that the hydrophilic dye Alexa 488 is imp

184 ch as projected cell area, adhesion area, or pipette size, as well as dynamical parameters such as th

185 the range approximately 0.4-4 mN/m, and the pipette size, one can precisely calculate the patch tens

186 ent was eliminated by removal of K+ from the pipette solution and partially blocked by the TASK (tand

187 against intracellular epitopes, in the patch pipette solution blocked TRPC1/C5 channel currents but p

188 caffolding domain peptide (10 microM) in the pipette solution completely abrogated the effects of ISO

189 With a pipette solution containing gramicidin, which forms Cl--

190 with a steeper dependence on voltage if the pipette solution contains K(+) as the main cation than i

191 as high and elimination of it with K(+)-free pipette solution could not be reconciled with restricted

192 rotons by buffering HC cytosol with a pH 9.2 pipette solution eliminated feedback, whereas alkalinizi

193 Perfusion of ADP or control pipette solution had no effect, whereas perfusion of ATP

194 clamping cytosolic Ca(2+) concentration with pipette solution in which Ca(2+) was buffered to 1 micro

195 robustly inhibits ClC-1 when included in the pipette solution in whole cell patch clamp experiments a

196 Inclusion of 1 mum Ins(1,4,5)P3 in the patch pipette solution increased whole-cell currents evoked by

197 (2-thiodiphosphate)] (2 mM) was added to the pipette solution of whole-cell recordings to regulate G

198 Recordings made with an ATP- and GTP-free pipette solution produced large and robust TRPC5 current

199 T cells with 200 muM Navbeta4 peptide in the pipette solution to induce resurgent sodium currents.

200 rat beta-cells by varying [Cl-] in the patch pipette solution using the Cl--permeable antibiotic amph

201 Increasing pH buffering capacity in the pipette solution with 40 mm HEPES attenuated the effects

202 ump) (both at 10 and 100 mmol/L Na(+) in the pipette solution) and maximal NKA-mediated Na(+) extrusi

203 alpha5alphaP is normally applied through the pipette solution, addition of steroid to the bath soluti

204 When 2 mM GDP-beta-S was added in pipette solution, ethanol-induced potentiation of I(Gly)

205 um through inclusion of 750 nm Ca(2+) in the pipette solution, indicating that neither the calcium se

206 When EGTA was omitted from the pipette solution, the number of sparks triggered in KO a

207 ing the driving force by modifying the patch-pipette solution.

208 and eliminated in the absence of ATP in the pipette solution.

209 te, whereas others used an ATP- and GTP-free pipette solution.

210 ut not other phosphoinositides, to the patch pipette solution.

211 the use of amphotericin B with a high [Cl-] pipette solution.

212 trical activity irrespective of [Cl-] in the pipette solution.

213 e, or a calmodulin inhibitory peptide in the pipette solution.

214 ng guanosine-5'-O-(2-thiodiphosphate) in the pipette solution.

215 dye from the cytoplasm with a dye-free patch pipette solution.

216 ould be reduced by inclusion of NH4Cl in the pipette solution.

217 Patch pipette solutions buffered with 1-4 mm of either BAPTA o

218 stabilizes when cells are patch clamped with pipette solutions containing 10 mm BAPTA and free Ca2+ c

219 tivity could be evoked by 16 mM glucose with pipette solutions containing 80 or 150 mM Cl-.

220 hole-cell current densities are similar with pipette solutions containing cesium, potassium, or sodiu

221 of it when the Na(+) concentration in patch pipette solutions perfusing the intracellular compartmen

222 With intracellular pipette solutions that controlled free [Ca(2+) ]i , we f

223 e cell patch clamp recordings were made with pipette solutions that support activation of both Ca2+-

224 ent when we included glutaredoxin 1 in patch pipette solutions to reverse glutathionylation.

225 ith 140 mM KCl and 1 mM Mg2+ in the bath and pipette solutions, two main open levels with conductance

226 gating were observed using MgATP or K2ATP in pipette solutions, which increases or decreases [Mg(2+)]

227 can produce currents similar to those in the pipette-spanning dome.

228 highly sensitive, and requires only a single pipetting step.

229 erformed at the start of the assay (i.e., no pipetting steps are necessary during the assay).

230 ny recent techniques still involve laborious pipetting steps for spheroid manipulation such as collec

231 technique significantly reduces the numerous pipetting steps of spheroid manipulation to a single pip

232 voirs situated on the microchip in only five pipetting steps using an 8-channel pipettor.

233 less sample and an order of magnitude fewer pipetting steps.

234 application of AMPH, via a whole-cell patch pipette, stimulated the trafficking of Y335A-hDAT.

235 that TRPV4 can be activated by tens of mm Hg pipette suctions with open probability rising with sucti

236 barium in the bath and by GTP-gamma-S in the pipette, suggesting activation of a G-protein inward rec

237 e and GTPgammaS introduced through the patch pipette, suggesting that the two photoreceptor types emp

238 ied steroid compensates for the diffusion of pipette-supplied steroid out of the patch to the rest of

239 Pipette-supported single particle electrodes may also fi

240 ne allows the GPMV contents to passivate the pipette surface, thereby dynamically blocking membrane s

241 In the present work we used patch pipette techniques to study the properties of a novel Ca

242 graphy scan was modified into a larger patch pipette that could be positioned with nanoscale precisio

243 rons, by using ionic conditions in the patch pipette that mimic those produced by AMPH stimulation, w

244 We describe quantum dot-coated glass pipettes that provide strong two-photon contrast at deep

245 With 5 mm Na+ in the pipette the reduction in release flux was greater (34 +/

246 roduce a Hewlett-Packard prototype picoliter pipette, the "thermal inkjet picofluidic system" (TIPS),

247 nce seal between a lipid bilayer and a glass pipette, the so-called "giga-seal", channel activity can

248 g steps of spheroid manipulation to a single pipetting; therefore, the errors from those steps are el

249 pepsin at known constant concentration were pipetted through an array of miniaturized chromatographi

250 d resilient; they self-heal when liquids are pipetted through them.

251 integrated the chemoenzymatic technique in a pipette tip (AutoTip) that was operated by an automated

252 omeric avidin-coated microbeads trapped in a pipette tip and has been used for genotyping single nucl

253 t attainable separation distance between the pipette tip and substrate surface ( approximately 1 nm)

254 rom mdx mice are strongly curved towards the pipette tip by actin pulling normal to the membrane.

255 ification that allows controlled increase of pipette tip diameter.

256 Based on these results, separations in micro-pipette tip format with these three types of stationary

257 f reverse-phase resin packed at the end of a pipette tip to quickly separate unreacted AdoMet from ra

258 Both solid-contact and "pipette tip"-based sensors were successfully applied to

259 suspended by an atomic-scale meniscus at the pipette tip, and to image their phase transformations wi

260 e actuator can rapidly shift the theta-glass pipette tip, thus exposing the target receptors to alter

261 We develop a micro-pipette tip-based nucleic acid test (MTNT) for high-thro

262 size approximately given by the size of the pipette tip.

263 MTNT consists of micro-pipette tips and embedded solid phase nucleic acid extra

264 nzyme activity was moderate; and (iii) C(18) pipette tips and solvent extraction, a method that enric

265 ak hydrostatic pressures, with liquid-filled pipette tips as fluid columns at the inlets, to introduc

266 StageTips are ordinary pipette tips containing very small disks made of beads w

267 tential is hampered by poor visualization of pipette tips in deep brain tissue.

268 By immersing pipette tips into Alconox, a commercially-available dete

269 hase polyamide resin (DPA-6S) in custom-made pipette tips.

270 with 1 mM fluo-3 and 3 mM EGTA via the patch pipette to buffer diadic cleft Ca2+, the number of spark

271 omised when BAPTA was added in the recording pipette to buffer intracellular Ca2+ and block the relea

272 light cures the thin adhesive layers linking pipette to collar to head.

273 Here, we used a focal pipette to deliver minimal stimulus shocks near second-o

274 ed in the internal solution of the recording pipette to reduce possible effects of network inhibition

275 For this purpose, calcium was pipetted to graphite furnace together with samples.

276 Using patch pipettes to record whole-cell currents under voltage cla

277 We used patch pipettes to record whole-cell currents under voltage-cla

278 The method involves using hand pipetting to create an array of cell-laden nanoliter-siz

279 We demonstrated the pipettes' utility in targeted patch-clamp recording expe

280 of 14 pA in 74% of macropatches attached to pipettes (-V(p) = -60 mV) containing a low-Na(+), nomina

281 esults (positive/negative) for WB sampled by pipette versus swab were concordant for 78/79 (98.7%) WB

282 ate on Xpert to compare WB sampling methods (pipette versus swab); 43/71 BS specimens had been previo

283 d the lowest tension is encountered near the pipette wall.

284 High-resolution TEM of carbon pipettes was used to attain better understanding of the

285 were displaced from the captured NA without pipetting wash buffers or use of external force and equi

286 rrents is fast and uniform before whole-cell pipette washout, suggesting little voltage attenuation a

287 muM calcium into the calyx via a whole-cell pipette, we found that slow endocytosis was inhibited by

288 tive or negative pressure from the recording pipette were considered to be transfected cells.

289 y stimulating the SNR close to the recording pipette, were inhibited to a smaller extent compared to

290 crease in [Na](i) at sites downstream of the pipette (where Na enters the myocyte and Na/K pumps are

291 ts have included ATP and/or GTP in the patch pipette, whereas others used an ATP- and GTP-free pipett

292 h ions were loaded through the somatic patch pipette, which also recorded electrical responses.

293 Here, we demonstrate the operation of a pipette, which, observed by transmission electron micros

294 We used dual pipette whole cell patch clamp recording to measure the

295 A1 pyramidal neurons were loaded via a patch pipette with a Ca(2+)-sensitive indicator (fura-6F) and

296 forces arising from a flow through a conical pipette with a tip radius of 1-1.5 mum, placed approxima

297 a servonull micropressure system, with glass pipettes with 2- to 3-microm tips used to cannulate epis

298 possibility of using nanometer-sized quartz pipettes with a layer of carbon deposited on the inner w

299 different, and, therefore, carefully chosen pipettes with well-characterized geometry were necessary

300 able, because variation in the amount of DNA pipetted would no longer contribute to variation in T/S,