Abstract

Single-cell measurements allow a unique glimpse into cell-to-cell heterogeneity; even small changes in selected cells can have a profound impact on an organism's physiology. Here an integrated approach to single-cell chemical sampling and assay are described. Capillary electrophoresis (CE) with laser-induced native fluorescence (LINF) has the sensitivity to characterize natively fluorescent indoles and catechols within individual cells. While the separation and detection approaches are well established, the sampling and injection of individually selected cells requires new approaches. We describe an optimized system that interfaces a single-beam optical trap with CE and multichannel LINF detection. A cell is localized within the trap and then the capillary inlet is positioned near the cell using a computer-controlled micromanipulator. Hydrodynamic injection allows cell lysis to occur within the capillary inlet, followed by the CE separation and LINF detection. The use of multiple emission wavelengths allows improved analyte identification based on differences in analyte fluorescence emission profiles and migration time. The system enables injections of individual rat pinealocytes and quantification of their endogenous indoles, including serotonin, N-acetyl-serotonin, 5-hydroxyindole-3-acetic acid, tryptophol and others. The amounts detected in individual cells incubated in 5-hydroxytryptophan ranged from 10 -14 mol to 10 -16 mol, an order of magnitude higher than observed in untreated pinealocytes.

Original languageEnglish (US)
Pages (from-to)2965-2972
Number of pages8
JournalAnalyst
Volume137
Issue number13
DOIs
StatePublished - Jul 7 2012

Fingerprint

Optical Tweezers
Single-Cell Analysis
Capillary electrophoresis
Capillary Electrophoresis
chemical analysis
electrokinesis
Lasers
fluorescence
Fluorescence
laser
Indoles
Chemical analysis
Serotonin
Catechols
Micromanipulators
Sampling
5-Hydroxytryptophan
lysis
Physiology
sampling

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Cite this

A hyphenated optical trap capillary electrophoresis laser induced native fluorescence system for single-cell chemical analysis. / Cecala, Christine; Rubakhin, Stanislav S.; Mitchell, Jennifer W.; Gillette, Martha U.; Sweedler, Jonathan V.

In: Analyst, Vol. 137, No. 13, 07.07.2012, p. 2965-2972.

Research output: Contribution to journalReview article

@article{ebdd6ed80dff42eeb914274ee4a176b8,
title = "A hyphenated optical trap capillary electrophoresis laser induced native fluorescence system for single-cell chemical analysis",
abstract = "Single-cell measurements allow a unique glimpse into cell-to-cell heterogeneity; even small changes in selected cells can have a profound impact on an organism's physiology. Here an integrated approach to single-cell chemical sampling and assay are described. Capillary electrophoresis (CE) with laser-induced native fluorescence (LINF) has the sensitivity to characterize natively fluorescent indoles and catechols within individual cells. While the separation and detection approaches are well established, the sampling and injection of individually selected cells requires new approaches. We describe an optimized system that interfaces a single-beam optical trap with CE and multichannel LINF detection. A cell is localized within the trap and then the capillary inlet is positioned near the cell using a computer-controlled micromanipulator. Hydrodynamic injection allows cell lysis to occur within the capillary inlet, followed by the CE separation and LINF detection. The use of multiple emission wavelengths allows improved analyte identification based on differences in analyte fluorescence emission profiles and migration time. The system enables injections of individual rat pinealocytes and quantification of their endogenous indoles, including serotonin, N-acetyl-serotonin, 5-hydroxyindole-3-acetic acid, tryptophol and others. The amounts detected in individual cells incubated in 5-hydroxytryptophan ranged from 10 -14 mol to 10 -16 mol, an order of magnitude higher than observed in untreated pinealocytes.",
author = "Christine Cecala and Rubakhin, {Stanislav S.} and Mitchell, {Jennifer W.} and Gillette, {Martha U.} and Sweedler, {Jonathan V.}",
year = "2012",
month = "7",
day = "7",
doi = "10.1039/c2an35198f",
language = "English (US)",
volume = "137",
pages = "2965--2972",
journal = "Analyst",
issn = "0003-2654",
publisher = "Royal Society of Chemistry",
number = "13",

}

TY - JOUR

T1 - A hyphenated optical trap capillary electrophoresis laser induced native fluorescence system for single-cell chemical analysis

AU - Cecala, Christine

AU - Rubakhin, Stanislav S.

AU - Mitchell, Jennifer W.

AU - Gillette, Martha U.

AU - Sweedler, Jonathan V.

PY - 2012/7/7

Y1 - 2012/7/7

N2 - Single-cell measurements allow a unique glimpse into cell-to-cell heterogeneity; even small changes in selected cells can have a profound impact on an organism's physiology. Here an integrated approach to single-cell chemical sampling and assay are described. Capillary electrophoresis (CE) with laser-induced native fluorescence (LINF) has the sensitivity to characterize natively fluorescent indoles and catechols within individual cells. While the separation and detection approaches are well established, the sampling and injection of individually selected cells requires new approaches. We describe an optimized system that interfaces a single-beam optical trap with CE and multichannel LINF detection. A cell is localized within the trap and then the capillary inlet is positioned near the cell using a computer-controlled micromanipulator. Hydrodynamic injection allows cell lysis to occur within the capillary inlet, followed by the CE separation and LINF detection. The use of multiple emission wavelengths allows improved analyte identification based on differences in analyte fluorescence emission profiles and migration time. The system enables injections of individual rat pinealocytes and quantification of their endogenous indoles, including serotonin, N-acetyl-serotonin, 5-hydroxyindole-3-acetic acid, tryptophol and others. The amounts detected in individual cells incubated in 5-hydroxytryptophan ranged from 10 -14 mol to 10 -16 mol, an order of magnitude higher than observed in untreated pinealocytes.

AB - Single-cell measurements allow a unique glimpse into cell-to-cell heterogeneity; even small changes in selected cells can have a profound impact on an organism's physiology. Here an integrated approach to single-cell chemical sampling and assay are described. Capillary electrophoresis (CE) with laser-induced native fluorescence (LINF) has the sensitivity to characterize natively fluorescent indoles and catechols within individual cells. While the separation and detection approaches are well established, the sampling and injection of individually selected cells requires new approaches. We describe an optimized system that interfaces a single-beam optical trap with CE and multichannel LINF detection. A cell is localized within the trap and then the capillary inlet is positioned near the cell using a computer-controlled micromanipulator. Hydrodynamic injection allows cell lysis to occur within the capillary inlet, followed by the CE separation and LINF detection. The use of multiple emission wavelengths allows improved analyte identification based on differences in analyte fluorescence emission profiles and migration time. The system enables injections of individual rat pinealocytes and quantification of their endogenous indoles, including serotonin, N-acetyl-serotonin, 5-hydroxyindole-3-acetic acid, tryptophol and others. The amounts detected in individual cells incubated in 5-hydroxytryptophan ranged from 10 -14 mol to 10 -16 mol, an order of magnitude higher than observed in untreated pinealocytes.

UR - http://www.scopus.com/inward/record.url?scp=84862173703&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84862173703&partnerID=8YFLogxK

U2 - 10.1039/c2an35198f

DO - 10.1039/c2an35198f

M3 - Review article

C2 - 22543409

AN - SCOPUS:84862173703

VL - 137

SP - 2965

EP - 2972

JO - Analyst

JF - Analyst

SN - 0003-2654

IS - 13

ER -