New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept
In parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventi...
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doaj-0a113a6804c94726946cdae3a89c6e212020-11-25T02:13:59ZengHindawi LimitedJournal of Biomedicine and Biotechnology1110-72431110-72512011-01-01201110.1155/2011/257060257060New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of ConceptKaren Nava-Castro0Romel Hernández-Bello1Saé Muñiz-Hernández2Galileo Escobedo3Jorge Morales-Montor4Departamento de Infectología e Inmunología Perinatal, Instituto Nacional de Perinatología, 11000 México DF, MexicoDepartamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP 70228, 04510 México DF, MexicoSubdirección de Investigación Básica, Instituto Nacional de Cancerología, Secretaría de Salud (SSA), 47000 México DF, MexicoDepartamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP 70228, 04510 México DF, MexicoDepartamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP 70228, 04510 México DF, MexicoIn parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventing evaluation of individual cells or specific cell types in a given parasite tissue or organ. Also, the extremely high interaction between helminthes and host cells (particularly immune cells) is an important point to be considered. It is really hard to obtain fresh parasites without host cell contamination. Then, it becomes crucial to determine that the analyzed proteins are exclusively from parasitic origin, and not a consequence of host cell contamination. Flow cytometry is a fluorescence-based technique used to evaluate the expression of extra-and intracellular proteins in different type cells, including protozoan parasites. It also allows the isolation and recovery of single-cell populations. Here, we describe a method to isolate and obtain purified helminthes cells.http://dx.doi.org/10.1155/2011/257060 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Karen Nava-Castro Romel Hernández-Bello Saé Muñiz-Hernández Galileo Escobedo Jorge Morales-Montor |
spellingShingle |
Karen Nava-Castro Romel Hernández-Bello Saé Muñiz-Hernández Galileo Escobedo Jorge Morales-Montor New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept Journal of Biomedicine and Biotechnology |
author_facet |
Karen Nava-Castro Romel Hernández-Bello Saé Muñiz-Hernández Galileo Escobedo Jorge Morales-Montor |
author_sort |
Karen Nava-Castro |
title |
New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept |
title_short |
New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept |
title_full |
New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept |
title_fullStr |
New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept |
title_full_unstemmed |
New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept |
title_sort |
new method to disaggregate and analyze single isolated helminthes cells using flow cytometry: proof of concept |
publisher |
Hindawi Limited |
series |
Journal of Biomedicine and Biotechnology |
issn |
1110-7243 1110-7251 |
publishDate |
2011-01-01 |
description |
In parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventing evaluation of individual cells or specific cell types in a given parasite tissue or organ. Also, the extremely high interaction between helminthes and host cells (particularly immune cells) is an important point to be considered. It is really hard to obtain fresh parasites without host cell contamination. Then, it becomes crucial to determine that the analyzed proteins are exclusively from parasitic origin, and not a consequence of host cell contamination. Flow cytometry is a fluorescence-based technique used to evaluate the expression of extra-and intracellular proteins in different type cells, including protozoan parasites. It also allows the isolation and recovery of single-cell populations. Here, we describe a method to isolate and obtain purified helminthes cells. |
url |
http://dx.doi.org/10.1155/2011/257060 |
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