Scanning cytometry with a LEAP

Laser-enabled analysis and processing of live cells in situ

Peter Szaniszlo, William A. Rose, Nan Wang, Lisa M. Reece, Tamara V. Tsulaia, Elie G. Hanania, Cornelis Elferink, James F. Leary

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: Scanning cytometry now has many of the features (and power) of multiparameter flow cytometry while keeping its own advantages as an imaging technology. Modern instruments combine capabilities of scanning cytometry with the ability to manipulate cells. A new technology, called LEAP™ (laser-enabled analysis and processing), offers a unique combination of capabilities in cell purification and selective macromolecule delivery (optoinjection). Methods: LEAP-mediated cell purification and optoinjection effects were assessed in model experiments using adherent and suspension cell types and cell mixtures plated and processed at different densities. Optoinjection effects were visualized by delivering fluorescent dextrans into cells. Results were analyzed using the LEAP instrument's own imaging system as well as by fluorescence and confocal microscopy. Results: Live cell samples (adherent and suspension) could be purified to 90-100% purity with 50-90% yield, causing minimal cell damage depending on the cell type and plating density. Nearly one hundred percent of the targeted cells of all cell types examined could be successfully optoinjected with dextrans of 3-70 kDa, causing no visual damage to the cells. Indirect optoinjection effects were observed on untargeted cells within 5-60 μm to targeted areas under conditions used here. Conclusions: LEAP provides solutions in cell purification and targeted macromolecule delivery for traditional and challenging applications where other methods fall short.

Original languageEnglish (US)
Pages (from-to)641-651
Number of pages11
JournalCytometry Part A
Volume69
Issue number7
DOIs
StatePublished - Jul 2006

Fingerprint

Lasers
Dextrans
Suspensions
Technology
Fluorescence Microscopy
Confocal Microscopy
Flow Cytometry
Cell Count

Keywords

  • Cell purification
  • Cytometry
  • Fluorescence microscopy
  • Laser-optoinjection
  • Scanning cytometry
  • Selective ablation
  • Selective delivery
  • Single cell analysis
  • Tissue surgery

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Szaniszlo, P., Rose, W. A., Wang, N., Reece, L. M., Tsulaia, T. V., Hanania, E. G., ... Leary, J. F. (2006). Scanning cytometry with a LEAP: Laser-enabled analysis and processing of live cells in situ. Cytometry Part A, 69(7), 641-651. https://doi.org/10.1002/cyto.a.20291

Scanning cytometry with a LEAP : Laser-enabled analysis and processing of live cells in situ. / Szaniszlo, Peter; Rose, William A.; Wang, Nan; Reece, Lisa M.; Tsulaia, Tamara V.; Hanania, Elie G.; Elferink, Cornelis; Leary, James F.

In: Cytometry Part A, Vol. 69, No. 7, 07.2006, p. 641-651.

Research output: Contribution to journalArticle

Szaniszlo, P, Rose, WA, Wang, N, Reece, LM, Tsulaia, TV, Hanania, EG, Elferink, C & Leary, JF 2006, 'Scanning cytometry with a LEAP: Laser-enabled analysis and processing of live cells in situ', Cytometry Part A, vol. 69, no. 7, pp. 641-651. https://doi.org/10.1002/cyto.a.20291
Szaniszlo P, Rose WA, Wang N, Reece LM, Tsulaia TV, Hanania EG et al. Scanning cytometry with a LEAP: Laser-enabled analysis and processing of live cells in situ. Cytometry Part A. 2006 Jul;69(7):641-651. https://doi.org/10.1002/cyto.a.20291
Szaniszlo, Peter ; Rose, William A. ; Wang, Nan ; Reece, Lisa M. ; Tsulaia, Tamara V. ; Hanania, Elie G. ; Elferink, Cornelis ; Leary, James F. / Scanning cytometry with a LEAP : Laser-enabled analysis and processing of live cells in situ. In: Cytometry Part A. 2006 ; Vol. 69, No. 7. pp. 641-651.
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abstract = "Background: Scanning cytometry now has many of the features (and power) of multiparameter flow cytometry while keeping its own advantages as an imaging technology. Modern instruments combine capabilities of scanning cytometry with the ability to manipulate cells. A new technology, called LEAP™ (laser-enabled analysis and processing), offers a unique combination of capabilities in cell purification and selective macromolecule delivery (optoinjection). Methods: LEAP-mediated cell purification and optoinjection effects were assessed in model experiments using adherent and suspension cell types and cell mixtures plated and processed at different densities. Optoinjection effects were visualized by delivering fluorescent dextrans into cells. Results were analyzed using the LEAP instrument's own imaging system as well as by fluorescence and confocal microscopy. Results: Live cell samples (adherent and suspension) could be purified to 90-100{\%} purity with 50-90{\%} yield, causing minimal cell damage depending on the cell type and plating density. Nearly one hundred percent of the targeted cells of all cell types examined could be successfully optoinjected with dextrans of 3-70 kDa, causing no visual damage to the cells. Indirect optoinjection effects were observed on untargeted cells within 5-60 μm to targeted areas under conditions used here. Conclusions: LEAP provides solutions in cell purification and targeted macromolecule delivery for traditional and challenging applications where other methods fall short.",
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