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Cell Biology JoVE (Open Access) Citable · DOI

Establishment of a Clonal Culture of Unicellular Conjugating Algae

DOI: 10.3791/57761-v
What you'll learn
  • ✓ Collect and identify unicellular conjugating algae from environmental water samples
  • ✓ Prepare glass capillary pipettes for single-cell micromanipulation
  • ✓ Isolate individual algal cells to establish clonal cultures
Protocol

Biopharma Insights In this article, we demonstrate the establishment of clonal cultures of unicellular conjugating algal species collected from a natural field site.

Difficulty
intermediate
Total time
~2–3 hours per isolation (including confirmation and microscopy)
Model organism
Unicellular conjugating algae (species from natural field collection)
Biosafety
BSL-1

Steps

1
Collect water sample containing algae

Obtain water samples from a natural field site containing unicellular conjugating algae. Process and prepare the sample for microscopic examination and isolation.

▶ 01:37
2
Confirm presence of target alga species

Examine the water sample under a microscope to identify and verify the presence of the unicellular conjugating algae of interest before proceeding to isolation.

▶ 02:56
3
Prepare glass capillary pipette from Pasteur pipette

Draw out a Pasteur pipette using heat to create a fine glass capillary with a narrow opening suitable for single-cell manipulation and isolation.

▶ 03:46
4
Isolate single algal cell using capillary pipette

Use the prepared glass capillary pipette under microscopic guidance to aspirate and transfer a single algal cell into fresh growth medium, establishing a clonal culture.

▶ 06:29
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