Isolation of Pneumocystis carinii cysts by flow cytometry

Jamie A. De Stefano; Richard G. Sleight; George F. Babcock; R. Michael Sramkoski; Peter D. Walzer

doi:10.1007/BF00931724

Isolation of Pneumocystis carinii cysts by flow cytometry

Jamie A. De Stefano, Richard G. Sleight, George F. Babcock, R. Michael Sramkoski, Peter D. Walzer

Periodontics

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2 Scopus citations

Abstract

Pneumocystis carinii cysts were separated as enriched populations from suspensions of lung homogenate obtained from infected rats containing all developmental stages of this organism. Isolation of cyst populations was achieved by incubating filtered lung homogenate with the fluorescent phospholipid analog 1-palmitoyl-2-C₆-NBD-phosphatidylcholine. Whereas cysts did not fluorescence as a result of outer-wall restraint of lipid integration, trophozoites and young intermediate stages readily incorporated the fluorescently labeled lipid analog into their outer membrane. The two distinct labeling patterns displayed by cysts and other developmental phases of P. carinii constitute a novel, easy, and reproducible means of isolating cysts from infected lung homogenate by flow cytometry.

Original language	English (US)
Pages (from-to)	179-182
Number of pages	4
Journal	Parasitology Research
Volume	78
Issue number	3
DOIs	https://doi.org/10.1007/BF00931724
State	Published - Mar 1992

ASJC Scopus subject areas

Parasitology
veterinary(all)
Insect Science
Infectious Diseases

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10.1007/BF00931724

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title = "Isolation of Pneumocystis carinii cysts by flow cytometry",

abstract = "Pneumocystis carinii cysts were separated as enriched populations from suspensions of lung homogenate obtained from infected rats containing all developmental stages of this organism. Isolation of cyst populations was achieved by incubating filtered lung homogenate with the fluorescent phospholipid analog 1-palmitoyl-2-C6-NBD-phosphatidylcholine. Whereas cysts did not fluorescence as a result of outer-wall restraint of lipid integration, trophozoites and young intermediate stages readily incorporated the fluorescently labeled lipid analog into their outer membrane. The two distinct labeling patterns displayed by cysts and other developmental phases of P. carinii constitute a novel, easy, and reproducible means of isolating cysts from infected lung homogenate by flow cytometry.",

author = "{De Stefano}, {Jamie A.} and Sleight, {Richard G.} and Babcock, {George F.} and {Michael Sramkoski}, R. and Walzer, {Peter D.}",

year = "1992",

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T1 - Isolation of Pneumocystis carinii cysts by flow cytometry

AU - De Stefano, Jamie A.

AU - Sleight, Richard G.

AU - Babcock, George F.

AU - Michael Sramkoski, R.

AU - Walzer, Peter D.

PY - 1992/3

Y1 - 1992/3

N2 - Pneumocystis carinii cysts were separated as enriched populations from suspensions of lung homogenate obtained from infected rats containing all developmental stages of this organism. Isolation of cyst populations was achieved by incubating filtered lung homogenate with the fluorescent phospholipid analog 1-palmitoyl-2-C6-NBD-phosphatidylcholine. Whereas cysts did not fluorescence as a result of outer-wall restraint of lipid integration, trophozoites and young intermediate stages readily incorporated the fluorescently labeled lipid analog into their outer membrane. The two distinct labeling patterns displayed by cysts and other developmental phases of P. carinii constitute a novel, easy, and reproducible means of isolating cysts from infected lung homogenate by flow cytometry.

AB - Pneumocystis carinii cysts were separated as enriched populations from suspensions of lung homogenate obtained from infected rats containing all developmental stages of this organism. Isolation of cyst populations was achieved by incubating filtered lung homogenate with the fluorescent phospholipid analog 1-palmitoyl-2-C6-NBD-phosphatidylcholine. Whereas cysts did not fluorescence as a result of outer-wall restraint of lipid integration, trophozoites and young intermediate stages readily incorporated the fluorescently labeled lipid analog into their outer membrane. The two distinct labeling patterns displayed by cysts and other developmental phases of P. carinii constitute a novel, easy, and reproducible means of isolating cysts from infected lung homogenate by flow cytometry.

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Isolation of Pneumocystis carinii cysts by flow cytometry

Abstract

ASJC Scopus subject areas

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