I am following recipe by Samson et al. 2014. They used agar and Tween 80 for spore suspension solution that was stored at 4 degree C. I do not understand how will I use agar containing solution in suspending spores. Kindly give insight. If possible, advice protocol of making spore suspension.
Conidia were harvested after cultivation for 7 days at 28 ± 0.5 ºC stirring the culture with a sterile 0.85% NaCl (Sigma–Aldrich St. Louis, MO, USA) solution containing 0.05% Tween 80 (Sigma–Aldrich St. Louis, MO, USA) for 5 min. The conidia suspension was gently probed with a pipette tip and filtered to separate conidia from hyphal fragments.
Barbara: Thanks for reply. I just wanted to ask that did you autoclave the NaCl+Tween 80 solution or did you filter sterilize it?
You can put a little of sterile Tween 20 in sterile water (a half or one drop of Tween 20 in a tube with 5ml of water or salina, for example). Add this solution on your Aspergillus culture, homogenize gently and return it again to tube. You may wait some minutes (~15 min) for the sedimentation of hyphae. You can confirm on microscopy exam. Take the suspension carefully, and add in another tube, for prepare your inocule test.
I agree with all above and you can see this link
http://www.microbiologyresearch.org/docserver/fulltext/micro/85/1/mic-85-1-173.pdf?expires=1481221451&id=id&accname=guest&checksum=A973A435AB50830F1A92524B83A2062D
Dear Rajshree Singh
Use a diluted aqueous solution of Tween 20® and place Aspergillus cultures. Stir well using a vortex. Microscopy and spectrophotometric studies will help you to determine the qualitative and features of your suspension.
Follow the RG link for relevant answers
https://www.researchgate.net/post/How_to_prepare_a_fungal_spore_suspension_from_a_plate_or_tube_for_later_inoculation_into_liquid_media
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