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Teaching Phagocytosis Using Flow Cytometry

    Authors: JOHN T. BOOTHBY1,*, RUTHANN KIBLER1, SABINE RECH1, ROBERT HICKS2
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    Affiliations: 1: Departments of Biological Sciences and; 2: Psychology, San Jose State University, One Washington Square, San Jose, California 95192-0100
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • *Corresponding author. Mailing address: Department of Biological Sciences, San Jose State University, One Washington Square, San Jose, CA 95192-0100. Phone: (408) 924-4850. Fax: (408) 924-4840. E-mail: jboothby@email.sjsu.edu.
    • Copyright © 2004, American Society for Microbiology
    Source: J. Microbiol. Biol. Educ. May 2004 vol. 5 no. 1 36-41. doi:10.1128/jmbe.v5.76
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    Abstract:

    Investigative microbiology on protists in a basic teaching laboratory environment is limited by student skill level, ease of microbial culture and manipulation, instrumentation, and time. The flow cytometer is gaining use as a mainstream instrument in research and clinical laboratories, but has had minimal application in teaching laboratories. Although the cost of a flow cytometer is currently prohibitive for many microbiology teaching environments and the number of trained instructors and teaching materials is limited, in many ways the flow cytometer is an ideal instrument for teaching basic microbiology. We report here on a laboratory module to study phagocytosis in sp. using flow cytometry in a basic microbiology teaching laboratory. Students and instructors found the flow cytometry data analysis program, Paint-AGate, to be very intuitive and easy to learn within a short period of time. Assessment of student learning about sp., phagocytosis, flow cytometry, and investigative microbiology using an inquiry-based format demonstrated an overall positive response from students.

Key Concept Ranking

Tetrahymena
0.44211397
Fluorescence Microscopy
0.4397621
Fluorescence Microscope
0.42695346
Light Microscopy
0.42350885
0.44211397

References & Citations

1. Bazzone DM1998An investigative approach to the study of phagocytosis inTetrahymena, p. 347–350 Karcher SJTested studies for laboratory teachingProceedings of the 19th Workshop/Conference of the Association for Biology Laboratory Education (ABLE) University of CalgaryAlberta, Canada
2. Colosi JC, Zales CR1998Jigsaw cooperative learning improves biology lab coursesBioScience4811812410.2307/1313137 http://dx.doi.org/10.2307/1313137
3. Davey HM, Kell DB1996Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analysesMicrobiol Rev606416968987359
4. Davis BB1993Tools for teachingJossey-Bass Publishers IncSan Francisco, Calif.
5. Gamson ZF1991A brief history of the seven principles for good practice in undergraduate education, p. 5–12 Chickering AW, Gamson ZFApplying the seven principles for good practice in undergraduate educationJossey-Bass Publishers, Inc.San Francisco, Calif.
6. Goodwin L, Miller JE, Cheetham RD1991Teaching freshmen to think—does active learning work?BioScience4171972210.2307/1311767 http://dx.doi.org/10.2307/1311767
7. Hatzis C, Srienc F, Fredrickson AG1994Feeding heterogeneity in ciliate populations: effects of culture age and nutritional stateBiotechnol Bioeng4337138010.1002/bit.26043050518615720 http://dx.doi.org/10.1002/bit.260430505
8. Hauer B, Eipel H1997Flow cytometry: useful tool for analyzing bacterial populations cell by cell Shapiro DBacteria as multicellular organismsOxford University PressNew York, N.Y.
9. Lavin DP, Fredrickson AG, Srienc F1990Flow cytometric measurement of rates of particle uptake from dilute suspensions by a ciliated protozoanCytometry1187588210.1002/cyto.9901108042125553 http://dx.doi.org/10.1002/cyto.990110804
10. Leonard WH2000How do college students best learn science?J Coll Sci Teaching24385388
11. Millis BJ, Cottell PGJr1998Cooperative learning for higher education facultyAmerican Council on Education/Oryx PressPhoenix, Ariz.
12. Talaro KP, Talaro A2002Foundations in microbiology4th edMcGraw-Hill, Inc.New York, N.Y.
13. Tortora GJ, Funk BR, Case CL2001Microbiology: an introduction7th edBenjamin Cummings, Inc.San Francisco, Calif.
14. Zhong Z, Pirofski LA1998Antifungal activity of a human antiglucuronoxylomannan antibodyClin Diagn Lab Immunol558649455881
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/content/journal/jmbe/10.1128/jmbe.v5.76
2004-05-01
2017-07-23

Abstract:

Investigative microbiology on protists in a basic teaching laboratory environment is limited by student skill level, ease of microbial culture and manipulation, instrumentation, and time. The flow cytometer is gaining use as a mainstream instrument in research and clinical laboratories, but has had minimal application in teaching laboratories. Although the cost of a flow cytometer is currently prohibitive for many microbiology teaching environments and the number of trained instructors and teaching materials is limited, in many ways the flow cytometer is an ideal instrument for teaching basic microbiology. We report here on a laboratory module to study phagocytosis in sp. using flow cytometry in a basic microbiology teaching laboratory. Students and instructors found the flow cytometry data analysis program, Paint-AGate, to be very intuitive and easy to learn within a short period of time. Assessment of student learning about sp., phagocytosis, flow cytometry, and investigative microbiology using an inquiry-based format demonstrated an overall positive response from students.

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Figures

Image of FIG. 1.

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FIG. 1.

Example of Worksheet 3 experimental question, design, and protocol.

Source: J. Microbiol. Biol. Educ. May 2004 vol. 5 no. 1 36-41. doi:10.1128/jmbe.v5.76
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FIG. 2.

Students observed a mixture of fluorescent-labeled and sp. under phase-contrast (A) and fluorescence (B) microscopy displayed on a 15-inch monitor via a video camera. Students identified the cells present and characterized them in terms of relative size (FSC), complexity (SSC), and fluorescence. Students were asked to predict the placement of each cell type (event) on dot plots of FSC versus SSC and fluorescence versus SSC. Example of a student plot where T = sp. and Y = yeast, fluorescent-labeled (C).

Source: J. Microbiol. Biol. Educ. May 2004 vol. 5 no. 1 36-41. doi:10.1128/jmbe.v5.76
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Image of FIG. 3.

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FIG. 3.

Students determined the percentage of sp. with ingested yeast over time. Students chose a sample time for analysis, opened an archived datafile for their time point in Paint-A-Gate, made two dot plots (FSC versus SSC and FL1-FITC yeast versus SSC), and painted and gated the sp. population (red events) (A). Students then painted the high FL1 events, representing the sp. cells with ingested yeast, on the right plot (SSC versus FL1-FITC yeast) green (B). Students compiled their results to determine the time it takes for sp. to ingest fluorescent-labeled yeast (C).

Source: J. Microbiol. Biol. Educ. May 2004 vol. 5 no. 1 36-41. doi:10.1128/jmbe.v5.76
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Image of FIG. 4.

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FIG. 4.

An example of student experimental results from investigating prey size preference (2-μm red fluorescent beads and 6-μm green fluorescent beads). sp. were fed fluorescent beads (inset) (A). Dot plots (side scatter versus forward scatter and green versus red fluorescence) show the distribution of sp. population (B). A graphical summary of student data is shown using the means of five to six replicas of each condition (C).

Source: J. Microbiol. Biol. Educ. May 2004 vol. 5 no. 1 36-41. doi:10.1128/jmbe.v5.76
Download as Powerpoint

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