TC contamination

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avadakedavra

 
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TC contamination

by avadakedavra » Tue May 23, 2017 11:35 pm

Hi, l have a question about Tissue culture of Cephalotus follicularis from its leaf.

I've started TC on 19th MAY and now I found some contamination in it.
Something white semitransparent looks like water mold are around some pieces of leaves.
It's not a solid colony of usual mold contamination.

Blank TC mediums are clear and no contamination, so I guess I failed to sterilize the samples of Cephalotus follicularis leaves.

The leaves are still fresh green. They are not dead yet.

May I try re-sterilize and set it again ?
or Keep watching a few days more ?

I have some ethanol, Hypochlorous acid solution and Sodium hypochlorite (as kitchen bleach)











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avadakedavra

 
Posts: 90
Joined: Thu May 18, 2017 3:06 pm
Thanked: 7 times in 7 posts

Re: TC contamination

by avadakedavra » Wed May 24, 2017 2:45 pm

avadakedavra wrote:Hi, l have a question about Tissue culture of Cephalotus follicularis from its leaf.

I've started TC on 19th MAY and now I found some contamination in it.
Something white semitransparent looks like water mold are around some pieces of leaves.
It's not a solid colony of usual mold contamination.

Blank TC mediums are clear and no contamination, so I guess I failed to sterilize the samples of Cephalotus follicularis leaves.

The leaves are still fresh green. They are not dead yet.

May I try re-sterilize and set it again ?
or Keep watching a few days more ?

I have some ethanol, Hypochlorous acid solution and Sodium hypochlorite (as kitchen bleach)




Sent from my iPhone using Tapatalk

Image

The contamination spreads in this half day.
So I decided to sterilize it again before give up.


Sent from my iPhone using Tapatalk

User Avatar
avadakedavra

 
Posts: 90
Joined: Thu May 18, 2017 3:06 pm
Thanked: 7 times in 7 posts

Re: TC contamination

by avadakedavra » Wed May 24, 2017 3:03 pm

avadakedavra wrote:
avadakedavra wrote:Hi, l have a question about Tissue culture of Cephalotus follicularis from its leaf.

I've started TC on 19th MAY and now I found some contamination in it.
Something white semitransparent looks like water mold are around some pieces of leaves.
It's not a solid colony of usual mold contamination.

Blank TC mediums are clear and no contamination, so I guess I failed to sterilize the samples of Cephalotus follicularis leaves.

The leaves are still fresh green. They are not dead yet.

May I try re-sterilize and set it again ?
or Keep watching a few days more ?

I have some ethanol, Hypochlorous acid solution and Sodium hypochlorite (as kitchen bleach)




Sent from my iPhone using Tapatalk

Image

The contamination spreads in this half day.
So I decided to sterilize it again before give up.


Sent from my iPhone using Tapatalk


Image
Image

I washed these pieces with running water well and put them into 100ppm Hypochlorous acid solution.
and soaked in 80% ethanol about 20 seconds.
Then put them into Hypochlorous acid solution again soaked about 10 minutes.

This sterilization may kill them. or not to enough for once contaminated.

I will write the result if it works well or not.

Comments and suggestions are always welcome.


Sent from my iPhone using Tapatalk

User Avatar
avadakedavra

 
Posts: 90
Joined: Thu May 18, 2017 3:06 pm
Thanked: 7 times in 7 posts

Re: TC contamination

by avadakedavra » Sun May 28, 2017 8:06 am

avadakedavra wrote:
avadakedavra wrote:
avadakedavra wrote:Hi, l have a question about Tissue culture of Cephalotus follicularis from its leaf.

I've started TC on 19th MAY and now I found some contamination in it.
Something white semitransparent looks like water mold are around some pieces of leaves.
It's not a solid colony of usual mold contamination.

Blank TC mediums are clear and no contamination, so I guess I failed to sterilize the samples of Cephalotus follicularis leaves.

The leaves are still fresh green. They are not dead yet.

May I try re-sterilize and set it again ?
or Keep watching a few days more ?

I have some ethanol, Hypochlorous acid solution and Sodium hypochlorite (as kitchen bleach)




Sent from my iPhone using Tapatalk

Image

The contamination spreads in this half day.
So I decided to sterilize it again before give up.


Sent from my iPhone using Tapatalk


Image
Image

I washed these pieces with running water well and put them into 100ppm Hypochlorous acid solution.
and soaked in 80% ethanol about 20 seconds.
Then put them into Hypochlorous acid solution again soaked about 10 minutes.

This sterilization may kill them. or not to enough for once contaminated.

I will write the result if it works well or not.

Comments and suggestions are always welcome.


Sent from my iPhone using Tapatalk

I've tried re-sterilization but it didn't work.
They all contaminated.

I set new TC of Cephalotus again.
When I have success I will write a new topic.


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