is i have a grid that 0.1mm and i count 30 yeast cells in a space of ten of them. the examined slurry was dilluted with 1/1 water...
how many cells are in a 70g slurry? i get 4.2 billion?
how many cells are in a 70g slurry? i get 4.2 billion?
quick question...about cell counting.
- Thread starter bracconiere
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Attached is an Excel spreadsheet I made. Apparently, just for you bracc. See attached.
DuncB
Well-Known Member
That is very neat, I have downloaded it, not sure I'll use it yet, wish there was a hobby microbiologist in the house.
The good thing about microbiologists is they are like biologists but a lot smaller.
The good thing about microbiologists is they are like biologists but a lot smaller.
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but you say to count only the blue cells! but there's no entry for the ones that stain blue with methylene blue, and thus viability! 
(and it gives me 10.5billion cells in my slurry....but it is only 70gs...and about about only 1/4" thick in the container....
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wait, that's me!!??
and this USB scope was only $15....it came with a little slide with a 10mm grid with 0.1mm deviations too, aparently for reasons that escape me the people that actually do this for reasons beyond a giggle, spend upwards of $150 on a hemocytometer? and my smear i have no idea how deep it was, which was something i was told not to touch in that spread sheet....(got one on the way though for $10
does have the 0.1mm well)
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Anyone who uses that spreadsheet (or similar) needs to pay close attention to all the parameters: the measuring area, the dilution factor (if used, but usually necessary), the various volumes that get you a good cells/volume number.
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i did! that why when i saw well depth of 0.1mm, i realized that me using my finger to smear it on this little sheet was probably going to be an underestimate....
DuncB
Well-Known Member
Well I was thinking need a microscope and realised that I had this on the shelf, I don't think that it's been used or the box opened in 40 years. Says 50, 100 and 200x.
40 years ago I was a small biologist I suppose.
Thought I was going to avoid cell counting, now I've got a microscope I just need the grid, running out of excuses fast. Ahhgh.
Although given the write up on the box I might be too busy to count yeast cells with 138 experiments to do.
40 years ago I was a small biologist I suppose.
Thought I was going to avoid cell counting, now I've got a microscope I just need the grid, running out of excuses fast. Ahhgh.
Although given the write up on the box I might be too busy to count yeast cells with 138 experiments to do.
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i don't think 200x is going to let you see yeast cells..you need like 1000x i believe?
(and a word of caution about these cheapie 1600x ones, it came with some shady software i didn't trust installing wanting me to grant it all kinds of rights to my system to work...could be an equivelent of a modern phising email, with more effort)
400x is the minimum IMO. Also, microscopy is ALL about the light. That scope above is not going to satisfy. You want a light source below the sample and a normal microscope condenser to focus the light on the stage.
Amscope has lots of scopes that are affordable. Clarity depends on the objective lenses, and that is a WHOLE other subject. Also, a X/Y stage is absolutely necessary to work at these magnifications. I'm sure Pastuer did not have them though, but he also did not have toilet paper. I think we have risen above that, n'est-ce pas?
Amscope has lots of scopes that are affordable. Clarity depends on the objective lenses, and that is a WHOLE other subject. Also, a X/Y stage is absolutely necessary to work at these magnifications. I'm sure Pastuer did not have them though, but he also did not have toilet paper. I think we have risen above that, n'est-ce pas?
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my cheapo has a light above the viewing area, but i've noticed it doesn't work that good. would turning the light off and maybe using it on a piece of white paper with light under it be better? or black paper, i've had the best luck so far with light brown table behind?
Well yes, to do any sort of yeast counting (and especially viability study) you must have light passing through the cells. I have zero experience with cheap scopes, so I can't recommend a workaround for you.
All light sources in the world are divergent. That is, they leave a point(ish) source and expand. But scopes with integrated light sources below the stage take that light and condense it with optics - the light's not diverging, it's converging on the tiny spot you're looking at. That's what you want. A condenser.
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so now i want to try it again with electrical tape around the clear part....maybe they just made it clear so people would be excited it lights up....
DuncB
Well-Known Member
@passedpawn
That little microscope does have a light below the table, can either angle the mirror to reflect sunlight in or angle it towards the unit to reflect the light from the lightbulb up.
I'm relieved that microscopes not up to the job, wonder if I can tempt anyone to " quickly count my yeast cells in the lab at the hospital ".
In fact just remembered there is a microscope at work in the MOHS lab only used 2 days a month, it's very fancy stereo etc. Now I'm going to have to look in yeast and see what I'm meant to be doing. I just thought the book was a virtue brewing book, I'd read Water but that was a bit dry, so couldn't face yeast.
Painting is all about the light as well I've heard. Never had a problem with a roller and a large brush though.
That little microscope does have a light below the table, can either angle the mirror to reflect sunlight in or angle it towards the unit to reflect the light from the lightbulb up.
I'm relieved that microscopes not up to the job, wonder if I can tempt anyone to " quickly count my yeast cells in the lab at the hospital ".
In fact just remembered there is a microscope at work in the MOHS lab only used 2 days a month, it's very fancy stereo etc. Now I'm going to have to look in yeast and see what I'm meant to be doing. I just thought the book was a virtue brewing book, I'd read Water but that was a bit dry, so couldn't face yeast.
OP
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@passedpawn what magnification would you say that is... it's the best the $15 scope will do, it won't focus at it's claim max of 1600x...those are 0.1mm squares.....and do i interpret that as right with the dye not staining the cells, they're alive?
or hell, they look kinda big... for something that should only be 10um....is that what i should be seeing?
No idea.
If those are yeast cells*, looks like about 200x. (* and that's a big IF because there's lots of other flotsam and jetsam in a beer sample). Yeast cells are typically about 5um (.005 mm). That means, if your squares were 0.1mm on a side, the cell should be 1/20th of that cell. So, either your understanding of your markings is wrong, or those aren't yeast. What are you using on your scope there, with the lines?
Here's what yeast cells look like under my scope. I took this image while drinking the beer it came from, Bells Oberon haha. BTW, the green arrow points at the only living cell in there (if you believe the stain).
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damn i appreciate the help, it's just the cheap little plastic "calibration" film that came with this thing.....
i put the plastic strip that says 0.1mm squares ontop of an upside down petri dish, and got this snap...looks way better! can't wait for the real hemocytometer to get here....
there for a bit i could even these the raises in the black grid paint...then it kinda got more blurry.....
well at any rate i'll see what i can do, but i did smear some yeast slurry underneath this...so what you're saying is those white bits at the top of that pic grouped together are the actual yeast?
I cannot confidently say anything about your images.
With a haemocytometer, you'll get something like this. Then we can talk
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i like the thin lines...but i want to point out those aren't 1/20th of the grid squares! (and this thing only captures in 640x480....those will just be white dots!)
looks more like 1/5th?
Look at the double-lines, compare to image I posted above in post #2. That's what I have there.
If you want to measure the cell count, you must be able to know the volume of your counting area. This means, there are lines, and you know the dimensions of the lines. It also means you have a known DEPTH - that is, you've filled an area that has a cover slip over it, and that area has a known depth. This is what a hemocytometer is good for.
If you want to measure the cell count, you must be able to know the volume of your counting area. This means, there are lines, and you know the dimensions of the lines. It also means you have a known DEPTH - that is, you've filled an area that has a cover slip over it, and that area has a known depth. This is what a hemocytometer is good for.
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should have it wed, or thur....type at ya again then!
Hemocytometers usually come with special thick cover slips that are specifically made to create the proper volume between the grid and the glass. My understanding is that normal coverslips are not created with this in mind.
We always had to use the special hemocytometer cover slips when counting cells in the lab for work. My use was always the large grids in the 4 corners because we were using human cells, which are far larger than yeast.
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well i got this glass 'thing'....finaly figered out i need to smush it with something.....i can kinda see the lines, and the cells...but with the light coming from above it's a white wash, hard to make out...anyone got an idea for like a little 4" LED flat light to light from underneath it? i looked but most are in the feet range....
(preferably cheap!)
(preferably cheap!
)
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https://www.ebay.com/itm/1337423886...B3k3aKAaDsGUFPYKZuNIBkl3iMul|tkp:BFBMoOPj7qlg
think i'll give it a go....
think i'll give it a go....
DuncB
Well-Known Member
@bracconiere
Having seen the above product, I wonder whether your phone with a white screen such as notes page or a picture of something really white might do it. You can adjust your screen brightness as well.
Having seen the above product, I wonder whether your phone with a white screen such as notes page or a picture of something really white might do it. You can adjust your screen brightness as well.
OP
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hate to say it, but now you make me feel old.. i've never had a cell phone...but yeah, that probably would work, if i had one.
DuncB
Well-Known Member
Got a laptop ? Notepad in windows is a nice even white background.
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no, but this fancy new monitor has 165hz refresh rate? i don't want to flip over though.....wait, but the reason i had to buy this, was because my old TV was dying....JUST maybe! thanks for the idea!
edit: nope, all i get is the RGB lines....
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You're still in the toy department.
The science dept is in the cellar... Apparently there's an entry fee.
DuncB
Well-Known Member
I did get the idea of using monitor with notepad on screen as the background to view xray films if an xray light box wasn't available published in a medical journal.
Those were the days using films, nowadays its all digital.
Those were the days using films, nowadays its all digital.
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DuncB
Well-Known Member
Not sure whether the haemocytometer would work vertical? There can't be much volume
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i layed my spare tv on it's back and used the scope on it....all i saw were the RGB lines from the screen....but it was promising, think the white light tracing thing will work way better then the light coming from the top....and YES @IslandLizard i'm about up to the cost of a digital $200 scope at this point! but having more fun doing it!
DuncB
Well-Known Member
CRT monitor would have that problem, white perspex / polythene in between screen and your slide?
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it's a flat panel...but yeah, a piece of paper would be a good idea....and i don't sweating on it, or as you say persperex over it? LOL
but damn it, i SOOOO amm going to learn how to count! anyone know where to get hemocytometer covers for cheap? this one didn't come with any? or what they're called so i can search for them?
I don't think that would fit the purpose for you either. Chances are optical resolution is still poor, not getting better with typically (highly) inflated "electronic" magnification levels. Small crap in, bigger crap out.
IMO, the only solution is a truly optical microscope with an appropriate condensor light. I've seen them on auction sites dealing in surplus lab equipment, coming from labs and colleges. It may take a while to find the right piece for your needs, and possibly around the same $200, I dunno.
But always know what you're buying, and whether it suits your purpose while it fits your budget!
OP
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somebody asked how long yeast stays fresh in the fridge? i was like, hum, need to do a cell count, some way to figure out how many are alive or dead after a certain amount of time.... now i have a $15 digital scope, a vial of methylene blue...a hemocytometer..... but having a blast! they always get you with the cheapie at first!
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yeah i can get an optical for $150 off ebay, with digital camera $200.... but to me this is a gag to see how i'd do a cell count if alcohol was illegal here....and i cared enough not just to buy fruit juice...
This is where I bought mine (site used to be cynmar.com). I paid $1.35 for a box in 2012 (which will last you forever). They are now $1.68. Inflation!
https://yoursciencehub.com/microscope-cover-slips-square-22-x-22mm.html
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