Hello again, I found some information you may find useful in regard to clearing dark chitinous specimens. The idea is to use a 10% solution of potassium hydroxide in a small mason jar or suitable heatproof glass container. Pour in just enough for your specimen and then put this jar into the centre of a small pan on the stove or hotplate and pour in some water around the jar to bring the level up to the same height as the KOH. In essence a double boiler. Turn on the heat and bring the water to a boil for 10 minutes. This will safely bring the temperature of the KOH to the same level as the water. Depending on the size of the specimen you could leave it a little longer. After the time is up take your specimen out of the KOH and transfer it to another vessel containing room temperature hydrogen peroxide (3-5%) for 20 to 30 minutes. Pour some 70% alcohol into another pan, bring to a boil and transfer the specimen from the hydrogen peroxide into it. There will be lots of bubbles escaping from the specimen which is what we want and this also neutralizes the KOH and H2O2 solutions. Again depending on specimen size this can take 5 - 10 mins or less. Once the bubbles stop, turn off the heat and let cool. Take out the specimen and prepare the same way as in these video's, ie: place on the slide with your shims, arrange and proceed from there. This method avoids the possibility of the specimen falling apart due to long soak times in the potassium solution. Here is a link that goes into a bit more detail. Take a look at 'table 1' further down the page: www.scielo.br/j/rbent/a/d5FjVV5Z6T8HTJbLkrcZ7Rw/?lang=en#
@tubamaxima187 Thank you so much for sharing this method. I have adapted your method for my Masters Capstone Project and would like to reference you properly and thank you! Is there a way to reach out to you?
To Cristianawaters3107: Thank you for your kind comment. I am glad my method is working for you. Yes, you can reach out to me: email: Ascott4277@gmail.com I would be interested in knowing what you do.
ceratin still is al hydophilic protein, so it is not necessary to use toxic xylol or pure ethanol. You can stay in hydrophilic mediums such as glyceringelatine. With KOH you make keratin more hydrohilic so its nonsense to pass specimens to hydrophobic mediums
@@tubamaxima187 The main effect of inhaling xylene vapor is depression of the central nervous system, with symptoms such as headache, dizziness, nausea and vomiting. At an exposure of 100 ppm, one may experience nausea or a headache. At an exposure between 200 and 500 ppm, symptoms can include feeling "high", dizziness, weakness, irritability, vomiting, and slowed reaction time. A condition called chronic solvent-induced encephalopathy, commonly known as "organic-solvent syndrome" has been associated with xylene exposure.
I appreciate your observation and comment but that has not been my experience. I would appreciate your examples for us to see so that we can make our own decisions.
@@lotharmayring6063 I agree, and stated, that Xylene is toxic, that is why I emphasized precautions. I only transfer slides from one container to the next container when I am outside in a well ventilated area. I use an activated charcoal face mask filter specifically designed for toxic vapors, acrylo-nitriile gloves to prevent skin contact and a powerful fan to blow away vapors. It's all in the video. Many things are forbidden because people do not use common sense. Is smoking cigarettes still allowed in Germany ?
Thanks ever so much for the educational video
Hello again, I found some information you may find useful in regard to clearing dark chitinous specimens. The idea is to use a 10% solution of potassium hydroxide in a small mason jar or suitable heatproof glass container. Pour in just enough for your specimen and then put this jar into the centre of a small pan on the stove or hotplate and pour in some water around the jar to bring the level up to the same height as the KOH. In essence a double boiler.
Turn on the heat and bring the water to a boil for 10 minutes. This will safely bring the temperature of the KOH to the same level as the water. Depending on the size of the specimen you could leave it a little longer.
After the time is up take your specimen out of the KOH and transfer it to another vessel containing room temperature hydrogen peroxide (3-5%) for 20 to 30 minutes.
Pour some 70% alcohol into another pan, bring to a boil and transfer the specimen from the hydrogen peroxide into it. There will be lots of bubbles escaping from the specimen which is what we want and this also neutralizes the KOH and H2O2 solutions. Again depending on specimen size this can take 5 - 10 mins or less. Once the bubbles stop, turn off the heat and let cool.
Take out the specimen and prepare the same way as in these video's, ie: place on the slide with your shims, arrange and proceed from there.
This method avoids the possibility of the specimen falling apart due to long soak times in the potassium solution.
Here is a link that goes into a bit more detail. Take a look at 'table 1' further down the page: www.scielo.br/j/rbent/a/d5FjVV5Z6T8HTJbLkrcZ7Rw/?lang=en#
Hi there, are you using pure xylene, or would xylene suffice? pure xylene seems hard to come by. Great videos!
I use xylene that I get at Home-Depot. It's real xylene, not sure how pure it is.
@tubamaxima187 Thank you so much for sharing this method. I have adapted your method for my Masters Capstone Project and would like to reference you properly and thank you! Is there a way to reach out to you?
To Cristianawaters3107:
Thank you for your kind comment. I am glad my method is working for you. Yes, you can reach out to me: email: Ascott4277@gmail.com I would be interested in knowing what you do.
Hello Sir... Good morning... Sir i want to prepare only antenna or legs....so this permanent sliding method also similar for preparing antenna or legs
Yes, the technique is the same.
Yes, the method is the same.
Thank you
That trump accent, love it!!!
ceratin still is al hydophilic protein, so it is not necessary to use toxic xylol or pure ethanol. You can stay in hydrophilic mediums such as glyceringelatine. With KOH you make keratin more hydrohilic so its nonsense to pass specimens to hydrophobic mediums
Sorry to disagree. I have tried many methods. My method works that is why I shared it on RUclips.
@@tubamaxima187 The main effect of inhaling xylene vapor is depression of the central nervous system, with symptoms such as headache, dizziness, nausea and vomiting. At an exposure of 100 ppm, one may experience nausea or a headache. At an exposure between 200 and 500 ppm, symptoms can include feeling "high", dizziness, weakness, irritability, vomiting, and slowed reaction time. A condition called chronic solvent-induced encephalopathy, commonly known as "organic-solvent syndrome" has been associated with xylene exposure.
I appreciate your observation and comment but that has not been my experience. I would appreciate your examples for us to see so that we can make our own decisions.
working with Xylols without exhauster ist suicide. And it can alo be absorbed from skin. In germany it is forbidden
@@lotharmayring6063 I agree, and stated, that Xylene is toxic, that is why I emphasized precautions. I only transfer slides from one container to the next container when I am outside in a well ventilated area. I use an activated charcoal face mask filter specifically designed for toxic vapors, acrylo-nitriile gloves to prevent skin contact and a powerful fan to blow away vapors. It's all in the video. Many things are forbidden because people do not use common sense. Is smoking cigarettes still allowed in Germany ?