Fluorescent Microscope Incubator DIY
Koz
I have an issue in need of some collective help.
I am starting an experiment for grad thesis that will involve
protein(GFP, RFP) expression during specific cell cycles.
I would observe this and track expression with some blob/color
detection algorithms I'm assembling in OpenFrameworks. Assumption is
that this will all work and have access to a fluorescent microscope.
On to the question:
Timelapse will last about 60 hours, and the cells(murine fibroblasts)
need to be in incubation + CO2, so placing them on the stage for the
duration is not going to work.
I do have a proper incubator that meets those requirements but I dont
think putting a microscope in a humid warm incubator is good for the
objectives.
Outside of specialized microscopy incubator, has anyone tackled or
have any advice in maintaing incubation, condition and avoiding
condensation?
Looking for a hack/creative solution, or alternatively, around NYC, if
anyone is willing to provide access.
I can provide access to lasercutter in exchange :)
I found a pretty cool solution but may be a bit too technical for the
amount of time I have.
http://www.popsci.com/diy/article/2011-10/video-smart-petri-dish-images-cells-using-smartphone-camera-and-legos
Thanks so much,
akoz
Nathan McCorkle
the incubator with the plate on top... a scan would be not be a single
moment snapshot, but its probably think enough to fit in the incubator
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Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics
naiverahim
John Griessen
> Timelapse will last about 60 hours, and the cells(murine fibroblasts)
> need to be in incubation + CO2, so placing them on the stage for the
> duration is not going to work.
How about an incubator "slab" with support gear outside connected by tubes
to keep it warm/humid, and a second chamber around the main one to make
a double pane window to lessen chances of condensation?
See sketch: http://ecosensory.com/diybio/stage_incubator-3.jpg
http://ecosensory.com/diybio/stage_incubator-3.gif
Simon Quellen Field
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Koz
thinking.
My thought was to use the regulation from the main incubator and pipe
the environment in.
However I was worried that if there are any cold spots, it would
result in condensation and a ruined image.
I see the circular flow is meant to avoid this.
Thanks, I'll start prototyping some things and see how it goes.
Koz
Jordan Miller
you can just get tanks of premixed 5% CO2 air that works great for
incubation microscopy on the cheap. co2 independent media usually has
negative effects which can include cell death.
jordan
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Koz
John Griessen
> Something like this?
> " http://www.warneronline.com/product_info.cfm?id=1362&name=CO2%20Microscope%20Stage%20Incubator%20(Okolab%20Electric)
Wow! Yes, that's it, and it seems to have an array of culture dishes to use at one go in the incubator.
So, concept is proven, simplify and cost reduce and it could be a diybio lab instrument...
Chris Templeman
John Griessen
> I see the circular flow is meant to avoid this.
even temps. Try to find out as much about the dimensions of the
Warner instruments stage so you can benefit from their
past developments. Search for what they have patented and how
long ago. Avoid using their current patented concepts.
On 03/06/2012 11:23 AM, Jordan Miller wrote:
> co2 controllers to mix the gas properly are many thousands of dollars.
But, you could make a CO2-stat fairly quickly and use a one time measurement
with a CO2 meter lab instrument to calibrate it every so often. The open loop
flow of CO2 is going to have problems of how do you get the humidity
just right as well as the CO2 balance, so it's not a cure all.
Simon Quellen Field
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Simon Quellen Field
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Chris Templeman
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John Griessen
> as your solution uses cheap microscopes slides why bother.
>
:-)
Talk is cheap.
;-)
Chris Templeman
AKS
I think what you are trying to do is to culture some cells and I assume
you are working on mammalian cells as you need CO2 buffer. I think if
you have access to syringe pump with programmable syringe then you
can get rid of CO2 buffer as long as you can warm up the cell culture
medium in syringe pump to be at 37.5 Degree Celcius. CO2 buffer is
for pH control and if you can set up a continuous cell culture medium
renewal, that buffer may be redundant. If you have access to microfluidics
then it is easy to fabricate PDMS (Silicone polymer) to develop micro-cell
culture/perfusion chips and they are very cheap to make. I hope this helps.
On 3/6/12, diy...@googlegroups.com <diy...@googlegroups.com> wrote:
> =============================================================================
> Today's Topic Summary
> =============================================================================
>
> Group: diy...@googlegroups.com
> Url: http://groups.google.com/group/diybio/topics
>
> - Fluorescent Microscope Incubator DIY [15 Updates]
> http://groups.google.com/group/diybio/t/99c67b6c7b1a0aa8
> - DIYBIO in the news..."Amateurs Are New Fear in Creating Mutant Virus" [5
> Updates]
> http://groups.google.com/group/diybio/t/b38161b7a6225115
> - ultracentrifuge [1 Update]
> http://groups.google.com/group/diybio/t/b9f495154f37a0c
> - is it possible to isolate a unknown compound through HPLC [1 Update]
> http://groups.google.com/group/diybio/t/7746ae58df706f14
> - Microbially Accelerated Carbon Sequestration [2 Updates]
> http://groups.google.com/group/diybio/t/93a74d0adb5543ae
> - EWAste [1 Update]
> http://groups.google.com/group/diybio/t/e9f593397c9a2cff
>
>
> =============================================================================
> Topic: Fluorescent Microscope Incubator DIY
> Url: http://groups.google.com/group/diybio/t/99c67b6c7b1a0aa8
> =============================================================================
>
> ---------- 1 of 15 ----------
> From: Koz <kozov...@gmail.com>
> Date: Mar 05 11:16PM -0800
> Url: http://groups.google.com/group/diybio/msg/e73af425bf92372f
>
> HI Everyone,
>
> I have an issue in need of some collective help.
> I am starting an experiment for grad thesis that will involve
> protein(GFP, RFP) expression during specific cell cycles.
> I would observe this and track expression with some blob/color
> detection algorithms I'm assembling in OpenFrameworks. Assumption is
> that this will all work and have access to a fluorescent microscope.
> On to the question:
> Timelapse will last about 60 hours, and the cells(murine fibroblasts)
> need to be in incubation + CO2, so placing them on the stage for the
> duration is not going to work.
> I do have a proper incubator that meets those requirements but I dont
> think putting a microscope in a humid warm incubator is good for the
> objectives.
> Outside of specialized microscopy incubator, has anyone tackled or
> have any advice in maintaing incubation, condition and avoiding
> condensation?
> Looking for a hack/creative solution, or alternatively, around NYC, if
> anyone is willing to provide access.
> I can provide access to lasercutter in exchange :)
>
> I found a pretty cool solution but may be a bit too technical for the
> amount of time I have.
> http://www.popsci.com/diy/article/2011-10/video-smart-petri-dish-images-cells-using-smartphone-camera-and-legos
>
> Thanks so much,
>
> akoz
>
>
> ---------- 2 of 15 ----------
> From: Nathan McCorkle <nmz...@gmail.com>
> Date: Mar 06 02:50AM -0500
> Url: http://groups.google.com/group/diybio/msg/d25266f0f36ec7c3
>
> Could you put filter over a scanner CCD, then just put the scanner in
> the incubator with the plate on top... a scan would be not be a single
> moment snapshot, but its probably think enough to fit in the incubator
>
>
> --
> Nathan McCorkle
> Rochester Institute of Technology
> College of Science, Biotechnology/Bioinformatics
>
>
> ---------- 3 of 15 ----------
> From: naiverahim <rahim...@gmail.com>
> Date: Mar 06 06:39AM -0800
> Url: http://groups.google.com/group/diybio/msg/274f598719c25256
>
> it seems like the lego is there to secure the sample. why not just get a
> block of Styrofoam and cut a whole in it to place a small petri dish w/
> cover in it. then just secure a smartphone over the petri dish, connect the
> phone to computer and do the same thing.
>
> rahim
>
> On Tuesday, March 6, 2012 2:16:54 AM UTC-5, Koz wrote:
>
>
> ---------- 4 of 15 ----------
> From: John Griessen <jo...@industromatic.com>
> Date: Mar 06 09:39AM -0600
> Url: http://groups.google.com/group/diybio/msg/98016614d42efbb6
>
> On 03/06/2012 01:16 AM, Koz wrote:
>> Timelapse will last about 60 hours, and the cells(murine fibroblasts)
>> need to be in incubation + CO2, so placing them on the stage for the
>> duration is not going to work.
>
> How about an incubator "slab" with support gear outside connected by tubes
> to keep it warm/humid, and a second chamber around the main one to make
> a double pane window to lessen chances of condensation?
>
> See sketch: http://ecosensory.com/diybio/stage_incubator-3.jpg
> http://ecosensory.com/diybio/stage_incubator-3.gif
>
>
> ---------- 5 of 15 ----------
> From: Simon Quellen Field <sfi...@scitoys.com>
> Date: Mar 06 08:20AM -0800
> Url: http://groups.google.com/group/diybio/msg/8a35c1ee32620145
>
> Something like this?
> "
> http://www.warneronline.com/product_info.cfm?id=1362&name=CO2%20Microscope%20Stage%20Incubator%20(Okolab%20Electric)
> "
>
> It appears you can buy the parts separately, and build what you need:
> "
> http://www.warneronline.com/product_info.cfm?name=Heated%20Top%20Coverslip%20for%20DH35i%20and%20DH-40i%20Chambers%20(HCS-1)&id=1213
> "
>
> "
> http://www.warneronline.com/product_info.cfm?id=1165&name=Micro-Incubation%20System%20(DH-40iL)
> "
>
> "
> http://www.warneronline.com/product_info.cfm?id=959&name=Culture%20Dish%20Incubator%20(DH-35iL)
> "
>
> The temperature controller and gas supplies can be DIY without much trouble.
>
> -----
> Get a free science project every week! "http://scitoys.com/newsletter.html"
>
>
>
>
>
>
> ---------- 6 of 15 ----------
> From: Koz <kozov...@gmail.com>
> Date: Mar 06 09:14AM -0800
> Url: http://groups.google.com/group/diybio/msg/f7df47d144500a4a
>
> That is a very interesting sketch and very similar to what I was
> thinking.
> My thought was to use the regulation from the main incubator and pipe
> the environment in.
> However I was worried that if there are any cold spots, it would
> result in condensation and a ruined image.
> I see the circular flow is meant to avoid this.
> Thanks, I'll start prototyping some things and see how it goes.
>
>
>
>
>
> ---------- 7 of 15 ----------
> From: Koz <kozov...@gmail.com>
> Date: Mar 06 09:22AM -0800
> Url: http://groups.google.com/group/diybio/msg/328088b40dbf8f39
>
> Well the phone is there only as a source of light/ shifting.
> There is a sensor at bottom to scan the culture and that needs to be
> networked in.
> Also the process involves burning off a layer from the ccd and afixing it
> in silicon and other such processes that I wouldn't have access to do
> reliably.
>
>
> On Tuesday, March 6, 2012 9:39:51 AM UTC-5, naiverahim wrote:
>
>
> ---------- 8 of 15 ----------
> From: Jordan Miller <jrd...@gmail.com>
> Date: Mar 06 12:23PM -0500
> Url: http://groups.google.com/group/diybio/msg/4d3e27dc60bd180c
>
> co2 controllers to mix the gas properly are many thousands of dollars.
> you can just get tanks of premixed 5% CO2 air that works great for
> incubation microscopy on the cheap. co2 independent media usually has
> negative effects which can include cell death.
>
> jordan
>
>
>
>
>
> ---------- 9 of 15 ----------
> From: Koz <kozov...@gmail.com>
> Date: Mar 06 09:23AM -0800
> Url: http://groups.google.com/group/diybio/msg/41fe3c15f567183a
>
> That is interesting.
> Thanks for the link!
>
> On Tuesday, March 6, 2012 11:20:44 AM UTC-5, Simon Field wrote:
>
>
> ---------- 10 of 15 ----------
> From: John Griessen <jo...@industromatic.com>
> Date: Mar 06 12:47PM -0600
> Url: http://groups.google.com/group/diybio/msg/2518601bcbc25dff
> ---------- 11 of 15 ----------
> From: Chris Templeman <christe...@gmail.com>
> Date: Mar 06 10:53AM -0800
> Url: http://groups.google.com/group/diybio/msg/d35fbbe75840e8ba
>
> Since you mentioned you had a laser cutter check out the
> openspectrometer.com guys and in particular the third video on their
> site. They show a pretty clever way to make 'cuvettes' or simple holders
> for the spectrometer, but the design would lend itself well to being scaled
> up a little bit to make a cell culture holder that you would mate to a
> microscope since it has two 'optical surfaces'. Further the C02 could be
> pipe in via larger holes then what they show in the movie.
>
> Chris
>
> On Tuesday, March 6, 2012 12:14:24 PM UTC-5, Koz wrote:
>
>
> ---------- 12 of 15 ----------
> From: John Griessen <jo...@industromatic.com>
> Date: Mar 06 12:57PM -0600
> Url: http://groups.google.com/group/diybio/msg/c03f1518fd42be1a
>
> On 03/06/2012 11:14 AM, Koz wrote:
>> I see the circular flow is meant to avoid this.
> Sure. Just flow enough to get turbulent stirring inside and
> even temps. Try to find out as much about the dimensions of the
> Warner instruments stage so you can benefit from their
> past developments. Search for what they have patented and how
> long ago. Avoid using their current patented concepts.
>
> On 03/06/2012 11:23 AM, Jordan Miller wrote:
> > co2 controllers to mix the gas properly are many thousands of dollars.
>
> But, you could make a CO2-stat fairly quickly and use a one time measurement
> with a CO2 meter lab instrument to calibrate it every so often. The open
> loop
> flow of CO2 is going to have problems of how do you get the humidity
> just right as well as the CO2 balance, so it's not a cure all.
>
>
> ---------- 13 of 15 ----------
> From: Simon Quellen Field <sfi...@scitoys.com>
> Date: Mar 06 11:17AM -0800
> Url: http://groups.google.com/group/diybio/msg/3412042949dc92da
>
> The top window should be glass, and should be 0.17 mm thick or less.
> Most microscope objectives are designed to work with glass coverslips
> of this thickness. See
> "
> http://www.microbehunter.com/2010/06/12/cover-glass-thickness-and-resolution/
> "
>
> An acrylic window 1/8 inch thick won't allow the objective to get close
> enough
> to even focus on the cells beneath at any decent magnification.
>
> The bottom window should be a normal microscope slide, especially if an oil
> immersion condenser is used.
>
> The trick of using double-stick tape is still quite useful -- but you don't
> need
> a laser cutter. Just cut the tape to the shape you want, stick it onto a
> slide,
> and stick a cover slip on top. The gas escape holes are simply cut into the
> tape, instead of into the top or bottom window. Scissors or an Xacto knife
> will do for cutting the tape.
>
> -----
> Get a free science project every week! "http://scitoys.com/newsletter.html"
>
>
>
>
> On Tue, Mar 6, 2012 at 10:53 AM, Chris Templeman
>
>
> ---------- 14 of 15 ----------
> From: Simon Quellen Field <sfi...@scitoys.com>
> Date: Mar 06 11:25AM -0800
> Url: http://groups.google.com/group/diybio/msg/581d4d032693d272
>
> The humidity may be easily finessed if the cells grow well at 100% humidity.
>
> Warm the air/CO2 to the desired temperature, bubble it through
>
> a warm water bath, and send it into the incubator cell. The temperature
> drop between the source and the incubator will keep the humidity at
> 100%.
>
> -----
> Get a free science project every week! "http://scitoys.com/newsletter.html"
>
>
>
>
>
>
> ---------- 15 of 15 ----------
> From: Chris Templeman <christe...@gmail.com>
> Date: Mar 06 11:53AM -0800
> Url: http://groups.google.com/group/diybio/msg/aff063b0fab577ea
>
> @Simon thanks for pointing out a cheaper way to achieve the same
> results...obviously you could use thinner acrylic...but as your solution
> uses cheap microscopes slides why bother.
>
> -Chris
>
> On Tuesday, March 6, 2012 2:17:53 PM UTC-5, Simon Field wrote:
>
>
>
> =============================================================================
> Topic: DIYBIO in the news..."Amateurs Are New Fear in Creating Mutant Virus"
> Url: http://groups.google.com/group/diybio/t/b38161b7a6225115
> =============================================================================
>
> ---------- 1 of 5 ----------
> From: Richard Proctor <richard...@gmail.com>
> Date: Mar 06 03:40AM -0800
> Url: http://groups.google.com/group/diybio/msg/a9db19a59451dd13
>
> #fearmongering
>
>
>
> ---------- 2 of 5 ----------
> From: Chris Templeman <christe...@gmail.com>
> Date: Mar 06 10:38AM -0800
> Url: http://groups.google.com/group/diybio/msg/ed909d362936356b
>
> @Richard I do agree that the article lends itself to fearmongering....
>
> A broader discussion that I think the article should have focused on is not
> whether DIY Biologists could make a deadly flu today or in the near future
> but whether Biologists of all types (amateur to professional) in general
> are coming to terms and taking a full account of the potential power they
> will have in shaping this world. It is somewhat of a subtle point I want
> to get across and forgive for choosing examples that are rife with baggage,
> but as someone with a physics background and not a bio background I can
> think of two examples to make my point. TNT and the Atomic Bomb. In both
> cases scientists came up with discoveries that have both well known
> positive and negative effects. Many lives were saved. Many lives were
> lost. What interests me is how the pioneers of these technologies dealt
> with their discoveries. Nobel, who invented TNT realizes that negative
> power he brought on this earth and starts the Nobel Prize to acknowledge
> and reward positive scientific progress. Einstein, a key person for atomic
> energy leading to the bomb, at first urges the US to make the bomb and once
> it is unleashed on the world spends the rest of his life actively working
> to free the world from Nukes. The technology was produced and after great
> negative incidents each man spent time and effort to attempt to rid the
> world of the negative aspects.
>
> We have not had the equivalent of TNT nor the Atomic Bomb with respect to
> synbio or other emerging biotech...should we start a discussion now or wait
> till it happens?
>
> What I am getting at and what I want to know is:
> 1. Are biologists aware of the great power they / may have in the near
> future? Are the pioneers of the new field of synbio really taking stock of
> what great power they hold?
> 2. If biologists are aware of the potential power who is making a point of
> saying "yes I know we will have great power. Some my use this power
> negatively, but here is why progress needs to continue and most importantly
> here is how we can overcome the negative aspects?
>
> I really want to know where this discussion is happening. Please point me
> to these scientists, leaders, etc... or maybe my understanding of biology
> is all wrong and large scale disasters are not possible...
>
> I think the article in the NYTimes is a continued back of forth of
> scientists debating the possibility of a particular person with a
> particular skill set doing something wrong, but what we really we should be
> seeing is a discussion and acknowledgement that 1) in the future biologists
> will wield an incredible amount of power (good and bad) and 2) how are
> these pioneers responding to that responsibility in the new world they are
> shaping?
>
> -Chris
>
>
>
> On Tuesday, March 6, 2012 6:40:03 AM UTC-5, Richard Proctor wrote:
>
>
> ---------- 3 of 5 ----------
> From: Bryan Bishop <kan...@gmail.com>
> Date: Mar 06 12:51PM -0600
> Url: http://groups.google.com/group/diybio/msg/8e6bbfee9e9a7b4a
>
> On Tue, Mar 6, 2012 at 12:38 PM, Chris Templeman
>
>> I really want to know where this discussion is happening. Please point me
>> to these scientists, leaders, etc... or maybe my understanding of biology
>> is all wrong and large scale disasters are not possible...
>
>
> In the archives. It's painful to keep rehashing these things. Also, I'm
> really disappointed by Jason in the article making it sound like everyone
> subscribed on this list has agreed to work under Jason's rules/oversight.
> Not cool... but the journalist probably just caught him off guard or
> something.
>
> - Bryan
> http://heybryan.org/
> 1 512 203 0507
>
>
> ---------- 4 of 5 ----------
> From: Chris Templeman <christe...@gmail.com>
> Date: Mar 06 11:13AM -0800
> Url: http://groups.google.com/group/diybio/msg/bc965d0b442c9a4d
>
> @Bryan, sorry it's painful. I guess I am a noob around these parts and I
> will check out the archives.
>
> However, in particular three quotes got really got to me (2 from prominent
> members of the DIYBio community) that I felt the discussion is warranted as
> those who are the 'public face' of DIY Bio aren't necessarily doing a great
> job of raising the conversation to where it should be, IMHO.
>
> 1. ...D.I.Y. biologists sometimes laugh at the sinister powers people
> think they have. “People overestimate our technological abilities and
> underestimate our ethics,” said Jason Bobe, a founder of
> DIYbio.org<http://diybio.org/>
> .
>
> 2. ...Todd Kuiken, a senior research associate at the Woodrow Wilson
> Center in Washington who specializes in the movement, points out that
> typical D.I.Y. projects are relatively simple, like inserting a gene into
> bacteria to make them glow. Producing viruses involves much more expensive
> equipment to do things like rearing host cells. “It’s not going to happen
> in someone’s basement,” he said.
>
> 3. ...Nor do these amateurs have the years of training it takes to grow
> viruses successfully. “It’s like I say, ‘I want to be a four-star chef,’ ”
> said Dr. Jorgensen, the president of Genspace, who worked with viruses for
> her Ph.D. “You can read about it, but unless someone teaches you side by
> side, I don’t think you’re going to get far.”
>
> -Chris
>
> On Tuesday, March 6, 2012 1:51:41 PM UTC-5, Bryan Bishop wrote:
>
>
> ---------- 5 of 5 ----------
> From: Cory Geesaman <co...@geesaman.com>
> Date: Mar 06 11:35AM -0800
> Url: http://groups.google.com/group/diybio/msg/b380bcac9c452aab
>
> For the second attempt (the new Google groups seems to be a bit glitchy
> when trying to post):
> From what I've heard of the mutH5N1 research it sounds like something that
> could feasibly be created with a medieval grasp of alchemy and an animal
> abuse case - and it probably applies to many more instances than just that
> particular virus since selective breeding is the tried and true method of
> engineering organisms for thousands of years. So yes, there seems to be an
> inherent risk involved - but it is damn foolish to even try it, from
> an amateur sense: I want to synthesize a Vaccine for Agamid Adenovirus -
> but am waiting until such time as I have a good lab to prevent the release
> of any form of the virus and one that will pass an EPA inspection. I might
> have tried with less precautionary measures in place if not posting about
> it in the DIYbio community and learning more about the subject from the
> discussion that came about - but that certainly isn't an indictment of the
> DIYbio group and it seems unfair to suggest that DIYbio could have any hand
> in this fear-mongering nonsense. The media has an agenda, it's best to
> just avoid them and ignore them until they go bankrupt.
>
> On Tuesday, March 6, 2012 1:38:26 PM UTC-5, Chris Templeman wrote:
>
>
>
> =============================================================================
> Topic: ultracentrifuge
> Url: http://groups.google.com/group/diybio/t/b9f495154f37a0c
> =============================================================================
>
> ---------- 1 of 1 ----------
> From: mad_casual <ademl...@gmail.com>
> Date: Mar 06 10:30AM -0800
> Url: http://groups.google.com/group/diybio/msg/329e8e3d8fb91472
>
> +1 for sanity. There have got to be dozens of YouTube videos of people
> using a dremel or other motor to explode CDs with ZERO PPE or other safety
> equipment. IMO, a bullet is the wrong analogy to draw to start with,
> bullets aren't lethal because of the kinetic energy they impart, cars and
> C4 tend to be more lethal strictly from KE. IIRC, TNT is ~4 kJ / g, so
> using your calculation, if the rotor ran for 30 s and released ALL of that
> energy simultaneously, you'd get about 1.5 g of TNT. For the right
> incentive, I'd detonate 1.5 g of TNT in my bare hand and I'd stand next to
> a styrofoam box while 1.5 g of TNT detonated inside with little to no
> incentive. Just like a grenade, the danger doesn't come from the transfer
> of kinetic energy, the danger comes from the shrapnel's ability to
> penetrate clothing/skin. A razor blade with paltry amounts of energy can
> kill. A razor sharp piece of plastic would require phenomenal amounts of
> both energy and design to penetrate even modest metal plating.
>
> On Sunday, March 4, 2012 8:39:10 PM UTC-6, Simon Field wrote:
>> speed of the very outside edge of the rotor. An inch in from the edge, you
>>
>> only get 200
>> meters per second.
>
> On Sunday, March 4, 2012 8:39:10 PM UTC-6, Simon Field wrote:
>
>
>
> =============================================================================
> Topic: is it possible to isolate a unknown compound through HPLC
> Url: http://groups.google.com/group/diybio/t/7746ae58df706f14
> =============================================================================
>
> ---------- 1 of 1 ----------
> From: mad_casual <ademl...@gmail.com>
> Date: Mar 06 09:49AM -0800
> Url: http://groups.google.com/group/diybio/msg/d3b86bfe23ac429d
>
> On Monday, March 5, 2012 5:46:12 AM UTC-6, debadutta bhoi wrote:
>> compounds are there, is it possible to isolate each compounds from the
>> crude extract through HPLC ?
>> please help
>
>
> More specifics would help. When you say 'crude extract' do you mean
> 'crushed or lysed plant cells in water' or do you mean 'salt precipitation
> and ethanol extraction of plant material that's been pushed through a .22
> um filter'? Crude extract meaning the extraction was poor/simple or meaning
> that it was a good/complex extraction just done by hand. Also, when you say
> 'each compounds' do you have something specific you're looking for or are
> you expecting an HPLC-detector system to point you in a direction. Because,
> depending on the detector system, you're going to get pointed in about 100
> different directions. As for actually doing the deed, you can easily
> overload an HPLC column, you can equally easily overload a detector. Given
> that you haven't overloaded the HPLC or the detector, then the answers spit
> out are always relative, especially for unknown complex mixtures. As a rule
> of thumb, 1000:1 relative concentrations are 'saturated'. Using a mass
> detector with a 1mL sample with 100 mg/mL of unkown solute, a competently
> designed modern system should be able to detect e.g. 1 mg of protein in 99
> mg of "stuff", etc. (~99:1). The well designed systems should be able to
> see 100-10 ug in a 100 mg sample (~1000:1). It's not too difficult to see
> low nano- to high pico- levels of compounds in a background of low
> micro-level contaminants. There are rather easy and even simple techniques
> for getting well beyond the 1000:1 level. Most LC-NMR and LC/LC-MS/MS/MS
> systems can be tailored to do this, bead based affinity systems can be done
> DIY. However, without knowing exactly what you're looking for, what Nathan
> says about getting into $100K+ instruments and Ph.D.-candidate level work
> is fairly cogent.
>
>
>
> =============================================================================
> Topic: Microbially Accelerated Carbon Sequestration
> Url: http://groups.google.com/group/diybio/t/93a74d0adb5543ae
> =============================================================================
>
> ---------- 1 of 2 ----------
> From: Nathan McCorkle <nmz...@gmail.com>
> Date: Mar 06 03:47AM -0500
> Url: http://groups.google.com/group/diybio/msg/90fa2093f931c683
>
> algae engineered to pump fatty acids out of their cells seems like it
> would be a winner, skim off the 'sequestrant', and throw it in the
> diesel car's fuel tank
>
>
> --
> Nathan McCorkle
> Rochester Institute of Technology
> College of Science, Biotechnology/Bioinformatics
>
>
> ---------- 2 of 2 ----------
> From: Mega <masters...@gmail.com>
> Date: Mar 06 08:22AM -0800
> Url: http://groups.google.com/group/diybio/msg/232676988b5a0449
>
> Nitrogen fixation.
>
> Nitrogen alone is no greenhouse gas. But if the bacteria bind nitrous
> oxides (which are 19-times more effecting in heating the Earth than
> crabon dioxide!) this would help against global warming.
>
>
>
> Anyway, in the more distant future we can't fix climate change only by
> removing carbon from the air. Plants need a specific level of carbon
> dioxide.
> Better remove more effective greenhouse gasses, like methane, water
> vapour, nitrous oxides, fluorcarbons (e.g. octafluorpropane), ....
>
>
>
> =============================================================================
> Topic: EWAste
> Url: http://groups.google.com/group/diybio/t/e9f593397c9a2cff
> =============================================================================
>
> ---------- 1 of 1 ----------
> From: Zebedee Boy <zebed...@hotmail.com>
> Date: Mar 06 11:38AM
> Url: http://groups.google.com/group/diybio/msg/c4cb40bafbe617
>
> It's an EU thing
> http://en.wikipedia.org/wiki/Waste_Electrical_and_Electronic_Equipment_Directive
> Dead electronics have to be recycled rather than landfilled as of 2007. Zeb
> From: jad...@azcobiotech.com
> To: diy...@googlegroups.com
> Subject: [DIYbio] EWAste
> Date: Mon, 5 Mar 2012 18:30:22 -0800
>
>
>
> Hi Ryan; Who is the eWaste company. I’ve never even heard of one. Thanks;
> J J AdamsAzco Biotech, Inc.3626 Ocean Ranch Blvd.Oceanside, CA 92056e.
> jad...@azcobiotech.comt. 858-259-9528m. 858-525-2770 From:
> diy...@googlegroups.com [mailto:diy...@googlegroups.com] On Behalf Of ryan
> martin
> Sent: Sunday, March 04, 2012 8:05 PM
> To: diy...@googlegroups.com
> Subject: [DIYbio] Equipment discussion I've been looking for second-hand lab
> hardware and have finally found an eWaste company that I can do business
> with. They have tons of equipment and I'm looking to start going over it and
> testing hardware to see what works and what doesn't. Centrifuges,
> microscopes, and the like I get; but when I get into analyzers and
> spectrometers I'll be having trouble figuring out testing protocols to
> ensure the hardware still functions (reliably). I'm looking for a good
> discussion list specifically for lab equipment testing, and maybe getting
> some help sorting out what might and might not be useful for a small scale
> diy bio lab. Thanks--
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>
>
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CodonAUG
Simon Quellen Field
Nathan McCorkle
> PDMS is widely used for microfluidics, but this particular application is
> not all
> that micro, and I really liked the double-stick tape method, as it is quick
> and
> very DIY accessible. Stick the tape on the slide, and use an Xacto knife to
> cut
> the channels and chambers, then apply the cover slip. Was that Nathan's
> idea,
> for the spectrometer cuvettes? I love it.
I think the sticky-tape cutout method was published previously, but my
roommate worked on a project last summer and he introduced me to it.
Zebedeeboy
Sent from Samsung Mobile
-------- Original message --------
Subject: Re: [DIYbio] Fluorescent Microscope Incubator DIY
From: Koz <kozov...@gmail.com>
To: diy...@googlegroups.com
CC:
That is interesting.
On Tuesday, March 6, 2012 11:20:44 AM UTC-5, Simon Field wrote:
Something like this?It appears you can buy the parts separately, and build what you need:The temperature controller and gas supplies can be DIY without much trouble.
On Tue, Mar 6, 2012 at 7:39 AM, John Griessen <jo...@industromatic.com> wrote:
On 03/06/2012 01:16 AM, Koz wrote:How about an incubator "slab" with support gear outside connected by tubes
Timelapse will last about 60 hours, and the cells(murine fibroblasts)
need to be in incubation + CO2, so placing them on the stage for the
duration is not going to work.
to keep it warm/humid, and a second chamber around the main one to make
a double pane window to lessen chances of condensation?
See sketch: http://ecosensory.com/diybio/stage_incubator-3.jpg
http://ecosensory.com/diybio/stage_incubator-3.gif
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