Counting chambers

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Rossf
Posts: 363
Joined: Mon Jan 20, 2020 12:39 am
Location: Victoria Australia

Counting chambers

#1 Post by Rossf » Fri Mar 12, 2021 12:49 am

Hello everyone-below is a pic of what seem to be fairly common counting chambers on eBay-how are they used? Is it just the see through windows where the solution goes then covered with a cover slip? They seem pretty thick-I was hoping thick enough to use 33mm rms objectives on stands that just don’t quite reach enough to focus! also does the solution take longer to drying out? Been frustrated filming microbes dividing (about half to full hour) but the big bubble of doom starts appearing before the quartering has completed. Do they have etched counting lines on the bottom? Without would be better.
Regards ross
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Placozoa
Posts: 92
Joined: Tue Jan 05, 2021 10:41 am

Re: Counting chambers

#2 Post by Placozoa » Fri Mar 12, 2021 1:16 am

https://www.ebay.com/itm/Microscope-obj ... Sw3ydV5aei

Maybe one of those would help? I bought one from that seller, it was metal, arrived fast, worked perfect.

Tom Jones
Posts: 336
Joined: Wed Apr 13, 2016 3:47 pm

Re: Counting chambers

#3 Post by Tom Jones » Fri Mar 12, 2021 1:38 am

It's called a Hemocytometer. They provide a flat, gridded and highly accurate counting volume. Before automated RBC, WBC, and platelet cell counters these are what was used to perform the counts in clinical labs. Lots of sperm counts and body fluid (pleural, peritoneal and CSF) cell counts are still done on them. Lots of other fluid particle counts too. Pretty easy to use, and you don't need any special objectives as you're only looking through a cover slip. See the links for procedures. For accurate counts you should use the reusable cover slips sold for the purpose. They're a little thicker than standard and insure the fluid volume is what it should be. For playing around, regular cover slips are probably fine. If you are trying to distinguish cell types, phase contrast is quite useful. It takes a fair amount of practice to get good. Even then, the coefficient of variation is pretty high.

https://en.wikipedia.org/wiki/Hemocytometer

https://incelligence.de/en/cell-culture ... g-protocol

dtsh
Posts: 977
Joined: Wed May 01, 2019 6:06 pm
Location: Wisconsin

Re: Counting chambers

#4 Post by dtsh » Fri Mar 12, 2021 2:58 am

Older ones come up somewhat often on eBay for very little and frequently come with micro pipettes. The coverglasses are not the typical slide coverglass, they're larger and thicker at 0.4mm. I'm pretty sure you can use standard coverglasses as the volume under the coverglass would be the same.

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Rossf
Posts: 363
Joined: Mon Jan 20, 2020 12:39 am
Location: Victoria Australia

Re: Counting chambers

#5 Post by Rossf » Fri Mar 12, 2021 4:28 am

Thanks people-Placozoa, I should just get some rms extenders-been meaning to get som fir ages-does it change working distance?
Regards ross

Tom Jones
Posts: 336
Joined: Wed Apr 13, 2016 3:47 pm

Re: Counting chambers

#6 Post by Tom Jones » Fri Mar 12, 2021 5:55 am

It doesn't change anything. It's just a thick glass base designed to be washed and reused many, many times. For years if you're careful. The distance to the floor of the counting chamber from the top of the cover slip is well within the working distance of regular 40x objectives. That is what they were designed for. I've done many hundreds of counts using them over the years. At least no one mouth pipets blood anymore to use them. :shock: I think I have five of them stashed away, several including the original cover slips and dilution pipets. :roll:

Tom Jones
Posts: 336
Joined: Wed Apr 13, 2016 3:47 pm

Re: Counting chambers

#7 Post by Tom Jones » Fri Mar 12, 2021 5:57 am

And they certainly do have etched grids for counting. Counts wouldn't be reasonably possible otherwise.

Stomias
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Joined: Tue Jan 12, 2021 7:25 pm

Re: Counting chambers

#8 Post by Stomias » Fri Mar 12, 2021 5:54 pm

Long ago I did WBC counts with them

Placozoa
Posts: 92
Joined: Tue Jan 05, 2021 10:41 am

Re: Counting chambers

#9 Post by Placozoa » Fri Mar 12, 2021 7:27 pm

Sorry, slightly different topic he was asking about. No, it doesnt change working distance. Slightly changes magnification but its not noticable, other than that it doesnt change anything. (Parfocal extenders)

chrisimbee
Posts: 105
Joined: Fri Mar 19, 2021 5:34 pm

Re: Counting chambers

#10 Post by chrisimbee » Fri Apr 16, 2021 9:01 pm

OMG ; i learnt counting blood cells with these !! and i even still have the special pipette to dilute blood with the coloring solution for th white cells !

its a Malasez counting cell but you need the special cover glass that goes with it :)
microscope Olympus BH2-BHTU+epifluo RFC @ 470 nm
Zeiss neofluar x16Ph, x40Ph, x100 oilPh
LOMO Ph x10 X20 X40 X90oil
Olympus SPFl x2
Olympus SPx20, SPx40 SPx100
camera astro ZWO ASI 120MM (n&b) et ZWO ASI 120 MC (colour)
Nikon D3100

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