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A comparison of techniques for introducing macromolecules into living cells
Journal article   Open access

A comparison of techniques for introducing macromolecules into living cells

Grace Lee, Thomas M. Delohery, Zeev Ronai, Paul W. Brandt‐Rauf, Matthew R. Pincus, Randall B. Murphy and I. Bernard Weinstein
Cytometry (New York, N.Y.), v 14(3)
1993
PMID: 7682492
url
https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/cyto.990140305View
Published, Version of Record (VoR) Open
url
https://doi.org/10.1002/cyto.990140305View
Published, Version of Record (VoR) Open

Abstract

Flow cytometry osmotic lysis of pinosomes red blood cell fusion scrape loading
Flow cytometry has been used to evaluate several techniques for introducing macromolecules into large numbers of living cells. One technique is cell fusion with red blood cell ghosts loaded with a fluorescent reporter molecule (RBCF). The second technique, termed osmotic lysis of pinosomes (OLP), involves a brief exposure of cells to a hypertonic solution containing the reporter molecule; subsequently, a hypotonic media is added which lyses the pinosomes formed during the hypertonic treatment. A third technique, scrape loading (SL), involves the creation of transient holes in the cell membrane through the application of mechanical forces, which allows for the passage of reporter molecules into cells. A comparison of these techniques is presented here. OLP appears to offer several advantages: It is a simple procedure, virtually all cells are fluorescently labelled, and it is capable of loading larger amounts of material more uniformly into cells while maintaining excellent viability. © 1993 Wiley‐Liss, Inc.

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Collaboration types
Domestic collaboration
Web of Science research areas
Biochemical Research Methods
Cell Biology
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