1887

Using Flow Cytometry to Measure Phagocytic Uptake in Earthworms

    Author: Sheryl L. Fuller-Espie1
    VIEW AFFILIATIONS HIDE AFFILIATIONS
    Affiliations: 1: Science Department, Cabrini College, Radnor, PA 19087
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 20 December 2010
    • Supplemental material available at http://jmbe.asm.org
    • *Corresponding author. Mailing address: Science Department, Cabrini College, 610 King of Prussia Road, Radnor, PA 19087-3698. Phone: (610) 902-8369. Fax: (610) 902-8285. E-mail: sfuller-espie@cabrini.edu.
    • Copyright © 2010 American Society for Microbiology
    Source: J. Microbiol. Biol. Educ. December 2010 vol. 11 no. 2 144-151. doi:10.1128/jmbe.v11i2.135
MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.
  • XML
  • HTML
    55.87 Kb
  • PDF
    237.22 Kb

    Abstract:

    This laboratory module familiarizes students with flow cytometry while acquiring quantitative reasoning skills during data analysis. Leukocytes, also known as coelomocytes (including hyaline and granular amoebocytes, and chloragocytes), from (earthworms) are isolated from the coelomic cavity and used for phagocytosis of fluorescent . Students learn how to set up cellular assays and become familiar with theoretical principles of flow cytometry. Histograms based on fluorescence and scatter properties combined with gating options permit students to restrict their analyses to particular subsets of coelomocytes when measuring phagocytosis, a fundamentally important innate immune mechanism used in earthworms. Statistical analysis of data is included in laboratory reports which serve as the primary assessment instrument.

Key Concept Ranking

Innate Immune System
0.471044
Polycyclic Aromatic Hydrocarbons
0.44493854
Chemicals
0.44211417
Reactive Oxygen Species
0.427141
Immune Response
0.42332727
0.471044

References & Citations

1. Axline SG, Reaven EP1974Inhibition of phagocytosis and plasma membrane mobility of the cultivated macrophage by Cytochalasin B: role of subplasmalemmal microfilamentsJ Cell Biol6264765910.1083/jcb.62.3.6474368822 http://dx.doi.org/10.1083/jcb.62.3.647
2. Cooper EL, Kauschke E, Cossarizza A2002Digging for innate immunity since Darwin and MetchnikoffBioEssays2431933310.1002/bies.1007711948618 http://dx.doi.org/10.1002/bies.10077
3. Engelmann P, Molnár L, Pálinkás L, Cooper EL, Németh P2004Earthworm leukocyte populations specifically harbor lysosomal enzymes that may respond to bacterial challengeCell and Tissue Res31639140110.1007/s00441-004-0874-x http://dx.doi.org/10.1007/s00441-004-0874-x
4. Forman HJ, Torres M2002Reactive oxygen species and cell signaling: respiratory burst in macrophage signalingAm J Respir Crit Care Med166S4S810.1164/rccm.220600712471082 http://dx.doi.org/10.1164/rccm.2206007
5. Fuller-Espie SL, Goodfield L, Hill K, Grant K, DeRogatis N2008Conservation of cytokine-mediated responses in innate immunity: a flow cytometric study investigating the effects of human proinflammatory cytokines on phagocytosis in the earthworm Eisenia hortensisInvertebrate Survival Journal5124134[Online] http://www.isj.unimore.it/
6. Givan AL2001Flow cytometry, first principles2nd ed1557John Wiley & Sons, Inc.New York10.1002/0471223948.ch3 http://dx.doi.org/10.1002/0471223948.ch3
7. Jarosz J, Glinski Z1997Earthworm immune responsesFolia Biologica4519
8. Malawista SE, Gee JB, Bensch KG1971Cytochalasin B reversibly inhibits phagocytosis: functional, metabolic, and ultrastructural effects in human blood leukocytes and rabbit alveolar macrophagesYale J Biol Med442863005132788
9. Segal AW2005How neutrophils kill microbesAnnu Rev Immunol2319722310.1146/annurev.immunol.23.021704.11565315771570 http://dx.doi.org/10.1146/annurev.immunol.23.021704.115653
10. Underhill DM, Ozinsky A2002Phagocytosis of microbes: complexity in actionAnnu Rev Immunol2082585210.1146/annurev.immunol.20.103001.11474411861619 http://dx.doi.org/10.1146/annurev.immunol.20.103001.114744
jmbe.v11i2.135.citations
jmbe/11/2
content/journal/jmbe/10.1128/jmbe.v11i2.135
Loading

Citations loading...

Supplemental Material

Loading

Article metrics loading...

/content/journal/jmbe/10.1128/jmbe.v11i2.135
2010-12-20
2017-09-23

Abstract:

This laboratory module familiarizes students with flow cytometry while acquiring quantitative reasoning skills during data analysis. Leukocytes, also known as coelomocytes (including hyaline and granular amoebocytes, and chloragocytes), from (earthworms) are isolated from the coelomic cavity and used for phagocytosis of fluorescent . Students learn how to set up cellular assays and become familiar with theoretical principles of flow cytometry. Histograms based on fluorescence and scatter properties combined with gating options permit students to restrict their analyses to particular subsets of coelomocytes when measuring phagocytosis, a fundamentally important innate immune mechanism used in earthworms. Statistical analysis of data is included in laboratory reports which serve as the primary assessment instrument.

Highlighted Text: Show | Hide
Loading full text...

Full text loading...

/deliver/fulltext/jmbe/11/2/jmbe-11-2-144.xml.a.html?itemId=/content/journal/jmbe/10.1128/jmbe.v11i2.135&mimeType=html&fmt=ahah

Figures

Image of FIGURE 1

Click to view

FIGURE 1

Shown are coelomocytes from after extrusion from the coelomic cavity. The large cells resembling fried eggs are the hyaline amoebocytes (large coelomocytes). The smallest cells with the white halo around their perimeter are the granular amoebocytes (small coelomocytes). The cells containing multiple small vesicular inclusions are the chlorogocytes (eleocytes). (Phase contrast microscopy, 400×)

Source: J. Microbiol. Biol. Educ. December 2010 vol. 11 no. 2 144-151. doi:10.1128/jmbe.v11i2.135
Download as Powerpoint
Image of FIGURE 2

Click to view

FIGURE 2

A forward (FSC) versus side (SSC) scatter profile of coelomocytes from . Note the rings [called regions (R)] drawn around the three major subpopulations: R1 = hyaline amoebocytes; R2 = granular amoebocytes; and R3 = chloragocytes. A region is a boundary drawn around a subpopulation for analysis. Once the regions have been established, analysis (e.g., FSC versus fluorescence detected by the FL-1 photomultiplier tube) can be carried out specifying which particular regions to include, a process known as gating. A gate may include one region or more than one region.

Source: J. Microbiol. Biol. Educ. December 2010 vol. 11 no. 2 144-151. doi:10.1128/jmbe.v11i2.135
Download as Powerpoint
Image of FIGURE 3

Click to view

FIGURE 3

Results of pre-test and post-test are illustrated. Left to right: pre-test for all students in class; post-test for all students in class; pre-test for students in class excluding students enrolled in BIO 312 (Theory & Practice in Biotechnology); post-test for students in class excluding students enrolled in BIO 312.

Source: J. Microbiol. Biol. Educ. December 2010 vol. 11 no. 2 144-151. doi:10.1128/jmbe.v11i2.135
Download as Powerpoint

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error