Question for Pipette users

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75RR
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Question for Pipette users

#1 Post by 75RR » Thu Oct 04, 2018 8:51 am

What would be the most useful volume sizes (µL) to use to transfer animalcules to slides?

Closing the barn door after the horse has already bolted of course ...

Just bought 3 lightly used variable volume Gilson P200, P1000 and P5000 which should arrive next week.

Range is:
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Re: Question for Pipette users

#2 Post by Hobbyst46 » Thu Oct 04, 2018 9:54 am

Hi 75RR,
Never done it, but intuitively, I would choose a 50ul tip, since an ordinary water drop is 0.05ml = 50ul volume.
But perhaps the inflow/outflow diameter also counts, the creature must pass through without crashing or clogging...
I can try and check with colleagues the exact inner diameter of the tip's end, but this will take a few days.
BTW - the Gilson Pipetman are excellent tools.
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Re: Question for Pipette users

#3 Post by 75RR » Thu Oct 04, 2018 10:07 am

Thanks Hobbyst46

was trying to visualize the quantities, good to know how much is in a drop of water ... looks like the P200 will be the most useful
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Re: Question for Pipette users

#4 Post by Hobbyst46 » Thu Oct 04, 2018 10:46 am

Another small point is that the typical volume of organic liquid drops is much smaller than that of water - drops of alcohols etc are about 20ul. This happens because of their lower surface tension.
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Re: Question for Pipette users

#5 Post by zzffnn » Thu Oct 04, 2018 12:50 pm

75RR,

When I worked in a microbiology lab, most of our modern P200 pipettes go down to 20 ul. That 20-200 up range is most useful for protist microscopy. Traditionally, P200 uses yellow tips for 20-200 ul, though some newer tips come in clear color now.

Out of your 3, I will suggest P200 for sure.

Most bio labs do not use P5000.

P20 can be useful too. Buy used or new/cheap ones from China. As hobbyists, we don't need volume precision. As long as it works well.

Edit: the only private pipette that I have at home is a P200 that goes down to 20 ul. It has a broken tip ejection system, so I bought it for $25 from eBay.

Look at the very tip of pipette tips under microscope and see what sizes work best. P20 with that traditional clear tip (for P10 and P20 that goes down to 2ul precisely) may precisely pick up a ciliate under stereo scope, if you are patient and lucky. For picking, those traditional yellow P200 tips may not be precise enough, but will still work.

Out of topic, when I was doing my internship for college about 17 years ago, I had a not-so-nice lab manager telling me "be careful with those pipettes, each of those may cost more than your TV". That may be true for very precise volume transfers, but we hobbyists don't need that kind of precision.

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Re: Question for Pipette users

#6 Post by 75RR » Thu Oct 04, 2018 1:29 pm

Thanks for the tips zzffnn
"be careful with those pipettes, each of those may cost more than your TV"
:)
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Re: Question for Pipette users

#7 Post by mnmyco » Fri Oct 05, 2018 10:16 pm

Gilson makes great pippetters. Pick whatever volume looks best abd cut the tips if you need a wider opening. Wouldn’t want to squish things through a tiny hole.

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Re: Question for Pipette users

#8 Post by charlie g » Fri Oct 05, 2018 10:35 pm

Hi 75RR..for collection and transfer of animicules...I find the protocol trumps pipet volume/pipet orifice size. I have a thread in here somewhere complete with images on this task.

Have three slides, each placed on a plastic condiment cup. These elevated slides permit a pen-light oblique illumination..in a dim or dark microscopy bench area..often your larger quarry (the meiofauna) can be naked eye observed with the pen light oblique illumination.


For the protists/protozoa...essential that your first sample from a culture or fresh collected sample be an elongated strip of fluid running 4/5ths the slide length. This crucial survey slide you scan at 4X, 10X ( often 20X objective possible with no coverslip...go for it!). When you observe target specimen you wish to transfer...simply alter your substage condenser to a tight light circle visible to you looking down at that elongated water drop (dim ight/ dark bench area always good for microscopy IMHO). Peek at your target specimen to insure it's 'in the spot-light'..then delftly pounce and slup that area of fluid up...place on the next slide..but as a more humble and symmetrical droplet.

Observe that droplet to verify you have your target organism...with practice you get it on 'the first slurp'...even if a below 40 micron target.

Go back to the initial scan slide...pursue more targets of interest, pounce and slurp...transfer as a humble symmetrical droplet adjacent but not confluent with that neighbor droplet holding a desired specimen.

Repeat this process till you have 'corralled a menagerie of target organisms' you dain worthy of the commitment of: dropping a coverslip upon for dedicated observations...first consider an agent to slow down motile organisms before you plop that cover slip ( You know 75RR I advocate use of viscosity tool methylcellulose 50/50 withyour droplet volume!).

If culture of corralled organisms (plant or animal as they say) is your objective..you still can benefit from this protocol. All microscopist of pond water/marine water live specimen observations should have a cheapo pen light at the bench, and try dark or at least dim light at bench to view organisms with oblique lillumination in their culture jars..with your pen light.

75RR..now that you made your purchase ( 'out of the barn already...ehh!?)..I insist you get back online an buy 'cheap!' : long neck and short neck borosilicate glass pastuer pipettes..with good 'rubber bulb reactive action'...these are all you need..it's the protocol...not the pipette..but borosilicate pastuer pipetts are essential at your bench..under your kitchen gas range stove...these pipettes easily draw out to micropipettes...but consider the 'harvest protocol first'. thanks for your microscopy, Charlie guevara

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Re: Question for Pipette users

#9 Post by 75RR » Sat Oct 06, 2018 3:33 am

75RR..now that you made your purchase ( 'out of the barn already...ehh!?)..I insist you get back online an buy 'cheap!' : long neck and short neck borosilicate glass pastuer pipettes..with good 'rubber bulb reactive action'...these are all you need..it's the protocol...not the pipette..but borosilicate pastuer pipetts are essential at your bench..under your kitchen gas range stove...these pipettes easily draw out to micropipettes...but consider the 'harvest protocol first'.
Thanks for the protocol charlie g - very useful
So, turn regular pipettes (with bulbs) into micro pipettes? That had not occurred to me. Can see why you mentioned cheap, suspect it will take a little practice. ;)

Looks like I will be selling the Gilson micropipettes on ...
( 'out of the barn already...ehh!?)..
Conditioned reflex ... made offer first then wondered how exactly I was going to use them!
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Re: Question for Pipette users

#10 Post by Hobbyst46 » Sat Oct 06, 2018 7:12 am

75RR wrote:Looks like I will be selling the Gilson micropipettes on ...
I think you will enjoy the Gilson dispensers. According to my experience, they are much more convenient than both the classical glass Pasteur pipettes and plastic disposable Pasteur pipettes for all dilutions, liquid exchange, addition of chemicals (to control pH, etc). Their high accuracy in this respect is much less important than the ergonomic advantage. In labs, the tips are packaged as arrays in special plastic boxes. One pushes the Pipetteman onto the tip, then proceeds to withdrawal and dispensing, then either disposal or cleaning and re-use. I prefer them to glass blowing any time - just my opinion. i do not know whether the Gilsons are also efficient for critter withdrawals.
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Re: Question for Pipette users

#11 Post by 75RR » Sat Oct 06, 2018 10:11 am

I prefer them to glass blowing any time - just my opinion.
Will play with them and see how it goes. Thanks
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Re: Question for Pipette users

#12 Post by 75RR » Thu Oct 11, 2018 6:28 pm

Well here they are. Look new compared to some used ones I have seen on ebay.
Surprisingly big as well. Largest is 24 cm not counting the plunger. Must say that they look and feel a little unwieldy.

Suppose I should give them a try. Will need to buy some tips, will these do for the P200?

https://www.ebay.co.uk/itm/Pipette-tips ... rk:19:pf:0
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Re: Question for Pipette users

#13 Post by Hobbyst46 » Thu Oct 11, 2018 7:32 pm

Shame on me for forgetting to verify!! Hopefully on Sunday I will contact some colleagues and inspect tip diameters.

Yet, I think that these 10ul tips will not fit on the P200 Pipeteman dispenser, since they are too small.

Also, the tips in the photos are packed "randomly". There are "racked tips", namely, packed tip arrays in special plastic dispensing boxes, in which the tips are parallel to each other and spaced, and are more convenient for routine use. With those, you just stick the Pipeteman into the tip, forcefully, and it is ready for use. They are probably more expensive, though.
There was a Gilson P20 Pipeteman dispenser as well. The 10ul tips would have fit on that one.
I should also add that tips are often incompatible with dispensers from a different manufacturers, because the taper angle is different or the shape is different, even when the dispensed liquid volume is the same.
Last edited by Hobbyst46 on Thu Oct 11, 2018 8:06 pm, edited 1 time in total.
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Re: Question for Pipette users

#14 Post by 75RR » Thu Oct 11, 2018 7:54 pm

Thanks Hobbyst46,
... you can put on a 200ul tip and adjust the volume to any level you like between 0 and 200ul.
Good to know. So the tip size for slurping up animalcules, as long as it is big enough for them to pass through, is not critical?

Then again a tip that is very large will probably siphon too much water along with the animalcule, defeating the purpose which is to isolate it.


Did not want to spend much money on the tips in case I decided that micropipettes were not for me.

Do not think I really need boxed tips either as a single tip will no doubt do for a whole stalking animalcules session.

Can see how having boxed tips is very time efficient in a commercial lab setting though. Less chance of cross contamination as well I imagine.
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Re: Question for Pipette users

#15 Post by Hobbyst46 » Thu Oct 11, 2018 8:09 pm

I edited my response somewhat, after some more thought.
I will try to check if other brand name tips will fit.
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Re: Question for Pipette users

#16 Post by 75RR » Thu Oct 11, 2018 8:21 pm

Thanks
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Re: Question for Pipette users

#17 Post by Hobbyst46 » Thu Oct 11, 2018 8:29 pm

Some links, two very inexpensive from China, appear to me OK but am not 100% sure that they fit the Gilson

1)
https://www.ebay.com/itm/1000-Pieces-Cl ... rk:14:pf:0

2)
https://www.ebay.com/itm/New-200pcs-200 ... rk:17:pf:0


And one more expensive, from USA, chance that the seller will know if the tip fits the Gilson P200
3)
https://www.ebay.com/itm/Extragene-Univ ... rk:19:pf:0
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Re: Question for Pipette users

#18 Post by 75RR » Thu Oct 11, 2018 8:44 pm

Thanks for that. Will ponder on it.
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Re: Question for Pipette users

#19 Post by 75RR » Thu Oct 11, 2018 8:52 pm

Found this from the same seller I referenced before. Ad states they will work on Eppendorf, Gilson, Socorex ...
5-200ul bag 1000 tips Universal(Yellow)
https://www.ebay.co.uk/itm/pipette-tips ... :rk:3:pf:0
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Re: Question for Pipette users

#20 Post by Hobbyst46 » Thu Oct 11, 2018 9:41 pm

Yes, I believe these should fit.
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Re: Question for Pipette users

#21 Post by mnmyco » Fri Oct 12, 2018 6:38 pm

They aren't unwieldy at all. If you find that you like a volume but not the tip opening, simply cut the tip and make the opening larger. Used to do it for certain protocols in a mycology lab. You should look up the info on servicing them, it is pretty easy. There are generally parts that are simply not servicable though and you need to know to not mess with those. Gilson are my one of my favorites really.

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Re: Question for Pipette users

#22 Post by 75RR » Fri Oct 12, 2018 8:54 pm

Thanks Hobbyst46 and mnmyco
If you find that you like a volume but not the tip opening, simply cut the tip and make the opening larger.
Good idea. That will make the 5-200µl tips more versatile.
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Re: Question for Pipette users

#23 Post by Hobbyst46 » Wed Oct 17, 2018 12:40 pm

75RR wrote:Thanks Hobbyst46 and mnmyco
If you find that you like a volume but not the tip opening, simply cut the tip and make the opening larger.
Good idea. That will make the 5-200µl tips more versatile.
Sorry for the delay. I have verified that two styles of tips fit the P-200 exactly. They are shown below. The one marked MT (in red) is from Mettler Toledo. The one marked G is original Gilson. Although the shapes are somewhat different, they both fit well. The photo was taken after I cut off the narrow tip, to verify the inner diameter. In the new, uncut tip, the entrance inner diameter is 600um (o.6mm) in both tips. On the original Gilson style, the taper is much more gradual along the whole length than in the MT style.
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Re: Question for Pipette users

#24 Post by 75RR » Wed Oct 17, 2018 1:09 pm

Thanks for that. Very useful info.
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Re: Question for Pipette users

#25 Post by Wes » Thu Mar 28, 2019 6:06 pm

I use Gilson pipettes on a daily basis, its like an extension of my hand now. The smallest volume you can pick up with the P200 is 20 ul as you know, however you can dispense as little as 1-2 ul simply by rotating the volumetric dial in small increments.

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Re: Question for Pipette users

#26 Post by charlie g » Thu Mar 28, 2019 7:37 pm

Hi, 75RR, you had a nice thread on this subject a few months ago. The process and protocol is the key issue all are silent on in this thread. You ask here:'the best volume pipet for transfer of animacules'.

So we are not concerned with solutions of bacteria mixed into a suspension, we are not concerned with precise volumes/ amounts of chemicals...I assume you are thinking of transfer of an individual target animalcule ? Perhaps I misread your meaning of the task of 'transfer of animacules'?

Do you mean: I observe a rotifer, a waterflea, a ciliate. diatom or desmid which I want to transfer for better control of a wetmount slide I intend to then image-capture as I observe the animalcules I fancy for that session of observation?

Or do you mean a pipet with orifice and volume suitable to 'slup up' animalcules whatever that pipet tip is encountering?

Different horses for different courses..pipets for serial high repetition accurate tasks do not lend themselves to handheld use at a wetdroplet slide on a microscope stage while observing a target animalcule. I'm not clear on your intent here with: 'transfer of animacules'.

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Re: Question for Pipette users

#27 Post by 75RR » Thu Mar 28, 2019 9:00 pm

Hi Wes and charlie g

Decided a while back that the Gilson pipettes weren't what I needed, they felt too bulky.

Sold them on and am now looking for some 5.25" Borosilicate Glass Pasteur Pipettes, but so far have not managed to locate any in Europe.

Came across some 9" but they seem much too long.

It was charlie g and his doggie that sold me on them. They are indispensable bear hunting tools. :)

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Re: Question for Pipette users

#28 Post by charlie g » Thu Mar 28, 2019 9:51 pm

Ooops..this the same thread started a while back?!! My bad. If all you find are the long pastuer pipets...if they are cheap/low cost..(they are cheap here in con.US..and lots available from China)...well buy these long ones and snap off the length you don't want...then 'fire polish at you kitchen gas-range stove.

A fast action rubber bulb plain eye dropper+ plus a pastuer pipet with a good seal/good action rubber bulb permits isolations, transfers, manipulations of animalcules..under 4X, even 10X objectives.

But all who enjoy live protist and meiofauna microscopy should aquire a 2X objective...manipulations easier with this near to macro-magnification objective.

happy springtime'19, Charlie guevara

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Re: Question for Pipette users

#29 Post by gastrotrichman » Fri Mar 29, 2019 3:20 am

I use and recommend Charlie's protocol, except that I use a stereo scope to make the captures from plastic Petri dishes and to set up the final slide for use with the compound scope. I heat and draw out the transfer pipette tip to increase water velocity (at least that's what I think happens) to more successfully suck up gastrotrichs, which typically attach tenaciously to the substrate upon the first hint of water movement. I use a rubber bulb with the pipette, but some people use flexible tubing to the user's mouth for suction.
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