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Laser Ablation of Lipofuscin

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#181 NasonSchooler

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Posted 22 December 2010 - 12:13 AM

I have put together a more formal proposal that is more specific as to where the money will go and what will be done on a week-by-week basis during the 4 month duration of the project.

Here is the proposal on Google Docs.


Share and enjoy!


Nason

#182 brokenportal

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Posted 27 December 2010 - 07:01 PM

Rather, I have pointed out that I am not in the area, and feel that any "oversight" worthy of the name is not possible remotely.

I have offered to make a statement that in my judgement there is no risk of unethical on incompetent behavior on Nason's part. I hereby make that statement.

This should not be construed as agreement to oversee the project in any way other than making the above statement.
However, if anyone has specific requests for me about what else I could do, I will be glad to hear them out.


Can we count you in to review Nasons bookwork once a month? I can see what you mean in that remote oversight might not be oversight at all. I agree with your statement that there is little to no risk of unethical or incompetent behavior. Do or dont call it oversight though, part of the idea would be to get more eyes on the data for potential (probable) added perspective.

#183 John Schloendorn

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Posted 28 December 2010 - 03:52 AM

You are certainly welcome to email and ask me stuff. I usually like to give technical comments on this or that. But I haven't received specific enough requests to make any specific promises.

#184 NasonSchooler

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Posted 16 January 2011 - 10:48 PM

I now have a check for supplies in hand - so I will start ordering stuff. First off are going to be plates and reagents to make NGM for the worms. Then I'll order the worms from CGC.


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#185 NasonSchooler

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Posted 26 January 2011 - 08:43 PM

Got plates and test tubes so far. Waiting for worms - should be here any day now...
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#186 NasonSchooler

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Posted 11 February 2011 - 03:53 PM

For some reason, my worm shipment from the CGC was returned undeliverable. I resent them my address and have my fingers crossed. If it doesn't work this time I'm driving up there to get the worms. Don't worry - the 4 month project does not officially start until I actually begin my first worm experiment. :)


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#187 AgeVivo

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Posted 13 February 2011 - 11:16 AM

just to give a feedback from a forum reader: not worried at all. in the contrary the devil is often in the details and it is nice that things advance. wishing all the best for this project
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#188 NasonSchooler

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Posted 16 March 2011 - 01:55 PM

Exp 1 should begin next week - I'm in the middle of building up worm stocks now.


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#189 NasonSchooler

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Posted 21 March 2011 - 07:13 PM

Started Experiment 1 today. My lab notes can be found here:
Lab Notes on Google Docs

3-21-2011, 2:00 PM
Started age-synchronized plates:
2 plates of N2, 40+ eggs each
2 plates of DA1116, 40+ eggs each
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#190 Mind

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Posted 28 March 2011 - 09:20 PM

I stopped by Nason's lab the other day to see how things are going and here are a few pictures of some of the equipment. This is the worm "hotel" - a temperature controlled refrigerator where the worms are grown and stored. Nason also has a heater in the research room to help keep the ambient temperature stable

Attached Files


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#191 Mind

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Posted 28 March 2011 - 09:26 PM

Here is Nason unpacking the laser, capable of producing 2 different wavelengths of light to test on the worms. Once things get rolling, he will have a jig or vice set-up to hold the laser at a standard distance from the worms to be treated. The laser is the same type of laser that would be used to remove tattoos or other cosmetic procedures on human skin.

Attached Files


Edited by Mind, 28 March 2011 - 09:35 PM.

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#192 Mind

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Posted 28 March 2011 - 09:30 PM

This is the bench where the treatments and analysis will be done. Also a picture of some of the first worms Nason has grown. I can't believe my wife got such a clear a picture of one of the round worms through the microscope lens with a point and shoot camera.

Attached Files


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#193 NasonSchooler

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Posted 30 March 2011 - 06:01 PM

Great photos, Justin. :)

Here's the latest update on what's going on in the lab:

Most of the eggs on the DA1116 plates were nonviable. I’ve been collecting all viable worms of this strain for breeding purposes. I went back and read up on the genetics a little bit, and the first few generations will have large numbers of nonviable offspring, since they are outcrossed with wild type and still have lots of homozygocity in many deleterious, recessive alleles. The outcrossing is designed to provide genetic diversity while keeping the desired traits. In a nutshell that means it will take awhile longer to get usable numbers of DA1116 worms. I will probably begin an N2-only experiment in the next few days, since I have the extra time.
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#194 AgeVivo

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Posted 04 April 2011 - 10:25 PM

great to see this progressive progress. we are all with you!
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#195 Elus

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Posted 05 April 2011 - 01:08 AM

We will finally get to see if the laser idea actually works. Can't believe it, fantastic work and keep it up!
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#196 NasonSchooler

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Posted 06 April 2011 - 01:37 AM

It's still taking awhile to breed more DA1116 worms. I can see how this is going to go - things are going to stretch out a bit, partly due to my schedule and partly due to using long-lived worms, and the nature of lifespan experiments in general. Therefore I propose using experiments as milestones instead of sticking to a fixed weekly or monthly schedule. Thus the project will span at least 8 complete lifespan experiments, regardless of how long it takes to complete them. The remaining 'monthly' salary and expense checks could be sent at the start of experiments 3, 5 and 7 - which will doubtless end up being more than one month apart. This definitely seems more appropriate to me - that way all of our gracious donors get the same amount of science for their money, regardless of how long it takes.

The fluorescence scope may or may not be purchased for this project, depending on how our financial situation pans out on this end. It may end up being budgeted as part of a future project proposal instead; but we can cross that bridge when we get there.

I'll let everyone know as soon as the worm zapping begins.


Nason
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#197 Mind

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Posted 11 April 2011 - 10:10 PM

Here is one of the short videos I shot will down in Iowa visiting Nason and the new lab. In it, Nason answers a question about the different types of lipofuscin that are found in tissue from different species and how the differences might affect the research.

Edited by Mind, 11 April 2011 - 10:11 PM.

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#198 Mind

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Posted 22 April 2011 - 06:53 AM

Here is a longer "basic" introduction/explanation to the experiment and the equipment that will be used (for those who are new to the proposal)

Edited by Mind, 22 April 2011 - 06:54 AM.

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#199 NasonSchooler

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Posted 26 April 2011 - 07:08 PM

Just wanted to let everyone know where I'm at - things got kind of busy on the home front, so I've had to postpone starting N2-only experiments. The mutants I've got are turning out to be too much hassle for doing every experiment with them. I think I'll wait until we've got something that looks promising as a treatment for the wild type, then include the caloric restriction mutants to see if the results are similar or different (either way). I plan to start up N2 experiments with at least 2 different treatment conditions per experiment (aside from controls) in a week or so - will let everyone know as soon as I get that up and running again.


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#200 treonsverdery

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Posted 26 April 2011 - 10:17 PM

New scientist has an article right now about chiral lasers are highly specific to particular molecules

It is possible that well tissue has chiral molecular tendencies whereas unwell tissue might have a different profile That strengthens the idea that laser modification of molecules could benefit tissue longevity

Reading Wikipedia suggests lipofuscin is a group of molecules rather than a specific I think many of these are chirally or possible circular polarizationally addressable

A super twisty beam of light has been created that can distinguish between left and right-handed molecules with unprecedented precision.
http://www.newscient...from-right.html

#201 NasonSchooler

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Posted 11 May 2011 - 01:00 AM

Cool!

#202 NasonSchooler

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Posted 11 May 2011 - 01:00 AM

I chunked worms today - I plan to get Exp 1 restarted this week...
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#203 brokenportal

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Posted 21 June 2011 - 07:49 PM

I chunked worms today - I plan to get Exp 1 restarted this week...


So how are things going now? I cant wait to find out how each stage turns out.

#204 NasonSchooler

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Posted 10 August 2011 - 09:14 PM

OK - the worms kept getting contaminated in my basement lab. We have spent the last month or so moving out of the bedroom and setting it up as a dedicated lab space that we can sterilize and keep that way. I will let everyone know when experiments are underway. We've also had trouble with time scheduling issues. I'm going to be babysitting during the day and doing experiments at night after the kids go to bed. It's the only way to give me sufficient time to dedicate to experiments. Sorry for all the delays, but I want to do this right.


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#205 NasonSchooler

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Posted 01 September 2011 - 04:16 PM

At Mind's suggestion, I have started a new forum topic for my current research involving sound/music vibration therapy.

#206 AgeVivo

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Posted 02 September 2011 - 09:34 AM

Do you need specific contacts for people who know specific techniques that you are a bit discovering?

In case you still try from time to time: I have worked with C elegans a little, I'm really not a pro but here are some tips against contamination: in a worm lab usually the microscope is on a glass surface that can be cleaned with alcohol very regularly, and on that glass surface, 5 to 10 cm away from the microscope, one uses a candle (oil-based candle generally, as it is rechargeable) that is lighted during worm manipulation (= opening the plates, sterilizing the metal stick in the flamme, transfering the worms from an old plate to a new plate, closing the new plate).
Have you used similar techniques? Briefly, how have you prepared the plates? (agar, feeding bacteria)

In case you don't try from time to time: are you going to send your equipment to a worm lab that might try the laser ablation of lipofuscin? Could it be a good idea to send it to someone from LongeCity willing to try his hands on C elegans, at home? If so, how much would it cost per month for that person to try to set up some lifespan experiment? Have you been in contact with people willing to try at home?
From far your idea of laser pulse to potentially increase C elegans lifespan still seems very interesting, mechanistically speaking at least.

Edited by AgeVivo, 02 September 2011 - 09:36 AM.


#207 seivtcho

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Posted 29 October 2011 - 10:57 AM

Please, write some more information about this project:

What laser will be used - wavelength, power, jauls per squaire centimeters
Over what cells / tissues / organs / or the whole body
Will it be over people or over animals, and which exactly animals
Do You have a methodics for the laser procedures or this has to be determined
Where will You get lipofuscine

I am asking that, because I have an access to two surgical lasers and two low power lasers for low energy treatment, and since I work with human individuals, I may eventually be of service and thus to save You some dollars by doing some of the needed things cheeper. Also, I know of a relatively cheap animal experimentation center, that will allow me to work with the animals and thus saving some money if the research must be done on animals. I am certified from the American laser association of laser dentistry and from the World Federation of Laser Dentistry, that allows me to use lasers for medical uses. I don't mind not to take money for my work and only for consumatives and for the tax for the usage of the lasers and taxes for other needs of the research.

#208 Mind

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Posted 29 October 2011 - 12:46 PM

Well, Nason stopped working on laser ablation to do other things. It was not a desirable outcome, considering we raised a lot of money for it. I take a lot of the blame for not better evaluating Nason's capabilities and desire to complete the research. When I visited and made the videos, everything seemed ok. My wife was more skeptical, and I should've listened to her. Venting here.

S123 talked to a professor at his University in Belgium and the professor said laser ablation would never work because there are cells in the gut that will absorb the same laser light that is absorbed by lipofuscin, and the gut cells will be destroyed (this is the condensed cliif-notes version) and the worms will die. Considering that when Nason actually did some minor work on this project when he was at the SENS RC, and the worms did not immediately die, I have not yet written laser ablation off 100%.

Seivtcho, you should talk to S123 to get the full explanation why his professor thinks the project would not work. Also, thanks so much for offering your expertise. If something does go forward with this, make sure to stay in touch with AgeVivo for tips on raising c.elegans. In fact, we have several members who have done research with worms that would be able to help you get set up.

Edited by Mind, 29 October 2011 - 12:49 PM.


#209 seivtcho

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Posted 29 October 2011 - 12:55 PM

I think that I understand what the professor at the Belgium university means.

It is really possible the power of the laser needed to destroy the lipofuscin to be
powerful enough to destroy the cell itself.

I'll try to do some prelimenary research (without being sponsored) to see if with
the two high power and the two low power lasers actually are cappable to destroy
lipofuscin and if the power neede is close to the ablation of the whole cell, then this
idea has a really big problem.

#210 s123

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Posted 29 October 2011 - 03:37 PM

S123 talked to a professor at his University in Belgium and the professor said laser ablation would never work because there are cells in the gut that will absorb the same laser light that is absorbed by lipofuscin, and the gut cells will be destroyed (this is the condensed cliif-notes version) and the worms will die. Considering that when Nason actually did some minor work on this project when he was at the SENS RC, and the worms did not immediately die, I have not yet written laser ablation off 100%.


His main point was that these fluorescent granules are NOT lipofuscin. He recently tested them (publication is still in progress) and found that they were NOT composed of lipofuscin.





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