Related Experiment Video
Updated: Apr 12, 2026
Protocol for Human Blastoids Modeling Blastocyst Development and Implantation
Published on: August 10, 2022
Features of Blastocystis spp. in xenic culture revealed by deconvolutional microscopy
Robyn Nagel1, Christian Gray, Helle Bielefeldt-Ohmann
1School of Veterinary Science, University of Queensland, Gatton, Queensland, Australia, robyn@tgclinic.com.au.
Abstract:
Blastocystis spp. are common human enteric parasites with complex morphology and have been reported to cause irritable bowel syndrome (IBS). Deconvolutional microscopy with time-lapse imaging and fluorescent spectroscopy of xenic cultures of Blastocystis spp. from stool samples of IBS patients and from asymptomatic, healthy pigs allowed observations of living organisms in their natural microbial environment. Blastocystis organisms of the vacuolated, granular, amoebic and cystic forms were observed to autofluorescence in the 557/576 emission spectra. Autofluorescence could be distinguished from fluorescein-conjugated Blastocystis-specific antibody labelling in vacuolated and granular forms. This antibody labelled Blastocystis subtypes 1, 3 and 4 but not 5. Surface pores of 1 μm in diameter were observed cyclically opening and closing over 24 h. Vacuolated forms extruded a viscous material from a single surface point with coincident deflation that may demonstrate osmoregulation. Tear-shaped granules were observed exiting from the surface of an amoebic form, but their origin and identity remain unknown.
Insights
Blastocystis parasites, linked to irritable bowel syndrome (IBS), show unique autofluorescence and surface pore activity. Researchers observed vacuolated forms extruding material, potentially for osmoregulation.
Area of Science:
- Microbiology
- Parasitology
- Cell Biology
Background:
- Blastocystis spp. are prevalent enteric parasites implicated in irritable bowel syndrome (IBS).
- Understanding Blastocystis morphology and behavior is crucial for diagnosing and treating associated conditions.
- Limited in-vivo observations hinder a complete understanding of Blastocystis life cycle and function.
Purpose of the Study:
- To investigate the morphology and behavior of living Blastocystis spp. in their natural microbial environment.
- To characterize the autofluorescence properties of Blastocystis organisms.
- To explore potential osmoregulatory mechanisms and other cellular processes in Blastocystis.
Main Methods:
- Utilized deconvolutional microscopy with time-lapse imaging for observing live Blastocystis cultures.
- Employed fluorescent spectroscopy to analyze autofluorescence emission spectra.
- Used Blastocystis-specific antibodies for immunofluorescence labeling of different subtypes.
Main Results:
- Observed autofluorescence in vacuolated, granular, amoebic, and cystic forms of Blastocystis (557/576 emission).
- Distinguished autofluorescence from antibody labeling; identified subtypes 1, 3, and 4 but not 5.
- Documented cyclic opening/closing of 1 μm surface pores over 24 hours and extrusion of material from vacuolated forms.
Conclusions:
- Autofluorescence serves as a distinguishing characteristic of Blastocystis organisms.
- Cyclic pore activity and material extrusion suggest active cellular processes like osmoregulation.
- Further research is needed to determine the origin and function of extruded granules from amoebic forms.
More Related Videos
08:28Morphometric Protocol for the Objective Assessment of Blastocyst Behavior During Vitrification and Warming Steps
Published on: February 28, 2019
12:11A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.
Published on: July 9, 2012