Jun 10, 2024
Staining Spores for Conidia Counting
- IAC 1,
- Sara de Oliveira Olimpio1,
- Rebecca Ogioni1
- 1Instituto Federal do Espírito Santos - Campus Alegre
Protocol Citation: IAC , Sara de Oliveira Olimpio, Rebecca Ogioni 2024. Staining Spores for Conidia Counting. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr8462lmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 14, 2024
Last Modified: June 10, 2024
Protocol Integer ID: 99758
Keywords: spores for conidia counting methylene blue, conidia counting methylene blue, conidia with the specific dye, staining spore, morphological identification of the microorganism, identification of microorganism, microorganism, conidia, specific dye, dye, morphological identification, bacteria, visualization of dead tissue
Abstract
Methylene blue has different functions depending on its concentration. Its use allows the visualization of dead tissues and bacteria, for example. As a solution, this dye allows the identification of microorganisms in general. For practical use in this matter, the morphological identification of the microorganism is obtained from the visualization of its conidia with the specific dye.
Procedure
Add sample in 1,5 mL tube;
Add 100 uL of methylene blue dye;
Homogenize by vortexing for 10 seconds and wait 2 minutes;
Centrifuge for 1 minute at 16.000g;
Pipette 50 uL into the Neubauer chamber or microscope slide and cover with a cover slip;
Evaluate under an optical microscope;
Store at 4ºC overnight before to use.
Materials
Tween® 80;
Methylene blue;
Distilled water.
Soluction Preparation (100 mL)
In a becker add 80 mL of distilled water;
Weigh 0.7g of methylene blue;
Pipette out 20 uL of Tween® 80;
Fill volume to 100 mL with distilled water.
References
Pham, Tuan Anh et al. “Production of Blastospore of Entomopathogenic Beauveria bassiana in a Submerged Batch Culture.” Mycobiology vol. 37,3 (2009): 218-24. doi:10.4489/MYCO.2009.37.3.218
Oktari, A., et al. "The bacterial endospore stain on Schaeffer Fulton using variation of methylene blue solution." Journal of Physics: Conference Series. Vol. 812. No. 1. IOP Publishing, 2017.