New Technique for High Resolution DNA Sizing in Epi-Illumination

J.M. Schins; A. Agronskaia; B.G. de Grooth; J. Greve

doi:10.1002/(SICI)1097-0320(19980601)32:2<132::AID-CYTO8>3.0.CO;2-N

New Technique for High Resolution DNA Sizing in Epi-Illumination

J.M. Schins, A. Agronskaia, B.G. de Grooth, J. Greve

Research output: Contribution to journal › Article › Academic › peer-review

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Abstract

We present a high-resolution DNA-sizing technique based on the principles of flow cytometry, using a high numerical aperture objective and epi-illumination. The new technique, designed for small fluorescing samples/particles (sub-micron diameter) suspended in a weakly fluorescent medium, makes use of an additional focus for high-precision particle localisation. This way, only those particles are considered that flow exactly through a well-defined volume. Results are presented for fluorescent beads, as well as for YOYO-stained plasmids containing 5,500 basepairs. The latter were measured with 6.2% resolution, setting a new limit to flow-based sizing of DNA.

Original language	English
Pages (from-to)	132-136
Number of pages	5
Journal	Cytometry
Volume	32
Issue number	2
DOIs	https://doi.org/10.1002/(SICI)1097-0320(19980601)32:2<132::AID-CYTO8>3.0.CO;2-N
Publication status	Published - 1998

Keywords

Particle trajectory
High numerical aperture
DNA-sizing
Additional focus

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10.1002/(SICI)1097-0320(19980601)32:2<132::AID-CYTO8>3.0.CO;2-N

Cite this

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title = "New Technique for High Resolution DNA Sizing in Epi-Illumination",

abstract = "We present a high-resolution DNA-sizing technique based on the principles of flow cytometry, using a high numerical aperture objective and epi-illumination. The new technique, designed for small fluorescing samples/particles (sub-micron diameter) suspended in a weakly fluorescent medium, makes use of an additional focus for high-precision particle localisation. This way, only those particles are considered that flow exactly through a well-defined volume. Results are presented for fluorescent beads, as well as for YOYO-stained plasmids containing 5,500 basepairs. The latter were measured with 6.2% resolution, setting a new limit to flow-based sizing of DNA.",

keywords = "Particle trajectory, High numerical aperture, DNA-sizing, Additional focus",

author = "J.M. Schins and A. Agronskaia and {de Grooth}, B.G. and J. Greve",

year = "1998",

doi = "10.1002/(SICI)1097-0320(19980601)32:2<132::AID-CYTO8>3.0.CO;2-N",

language = "English",

volume = "32",

pages = "132--136",

journal = "Cytometry",

issn = "0196-4763",

publisher = "Wiley",

number = "2",

}

TY - JOUR

T1 - New Technique for High Resolution DNA Sizing in Epi-Illumination

AU - Schins, J.M.

AU - Agronskaia, A.

AU - de Grooth, B.G.

AU - Greve, J.

PY - 1998

Y1 - 1998

N2 - We present a high-resolution DNA-sizing technique based on the principles of flow cytometry, using a high numerical aperture objective and epi-illumination. The new technique, designed for small fluorescing samples/particles (sub-micron diameter) suspended in a weakly fluorescent medium, makes use of an additional focus for high-precision particle localisation. This way, only those particles are considered that flow exactly through a well-defined volume. Results are presented for fluorescent beads, as well as for YOYO-stained plasmids containing 5,500 basepairs. The latter were measured with 6.2% resolution, setting a new limit to flow-based sizing of DNA.

AB - We present a high-resolution DNA-sizing technique based on the principles of flow cytometry, using a high numerical aperture objective and epi-illumination. The new technique, designed for small fluorescing samples/particles (sub-micron diameter) suspended in a weakly fluorescent medium, makes use of an additional focus for high-precision particle localisation. This way, only those particles are considered that flow exactly through a well-defined volume. Results are presented for fluorescent beads, as well as for YOYO-stained plasmids containing 5,500 basepairs. The latter were measured with 6.2% resolution, setting a new limit to flow-based sizing of DNA.

KW - Particle trajectory

KW - High numerical aperture

KW - DNA-sizing

KW - Additional focus

U2 - 10.1002/(SICI)1097-0320(19980601)32:2<132::AID-CYTO8>3.0.CO;2-N

DO - 10.1002/(SICI)1097-0320(19980601)32:2<132::AID-CYTO8>3.0.CO;2-N

M3 - Article

SN - 0196-4763

VL - 32

SP - 132

EP - 136

JO - Cytometry

JF - Cytometry

IS - 2

ER -

New Technique for High Resolution DNA Sizing in Epi-Illumination

Abstract

Keywords

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