Best counting chamber, micropipette

[Microfan]

Hello. i've searched the forum for advice regarding an affordable, good quality counting chamber or hemocytometer.. i'm interested in sperm counts. The reviews online are highly variable: from one to five stars for the same product! I am not using it professionally, so would prefer something around $100 if it's of reasonably good quality. Also, what size micropipette would be appropriate to make loading the chamber easier? i need a micropipette for 50mcg for another application anyway but would hope to purchase only one. Thanks for any suggestions.

[dtsh]

Hello. i've searched the forum for advice regarding an affordable, good quality counting chamber or hemocytometer.. i'm interested in sperm counts. The reviews online are highly variable: from one to five stars for the same product! I am not using it professionally, so would prefer something around $100 if it's of reasonably good quality. Also, what size micropipette would be appropriate to make loading the chamber easier? i need a micropipette for 50mcg for another application anyway but would hope to purchase only one. Thanks for any suggestions.

They pop up on eBay fairly often, a lot of old Spencer Bright-Line, Adams, etc. I've bought a few over the years for $10-$20, usually include mouth pipettes and misc parts (can't recommend using the mouth pipettes, but can add a bulb or syringe). Make sure whatever you get includes a coverslip as they are not the typical coverslip, instead being not only larger but thicker too (0.4mm).
Also, make sure whatever objectives you intend to use have enough working distance to reach all the way through the 0.4mm coverslip and into the chamber.

No advice on the micropipette, though curious to hear the replies.

[photomicro]

As suggested, second-hand ones are cheap as chips, and will be perfectly fine.

A standard 'Pasteur' pipette, glass or plastic is fine, as this is just to load the sample, not measure it. You have to load an excess anyway.

Also as stated, the correct thickness of coverslip is crucial, as this is how a suitable volume is arrived at, the depth of the sample being the all important variable. The grid itself is a known size, but the depth depends on how well the cover is mounted.

Using a haemocytometer to get valid results for counting a number of things is often covered in text books.

There have been some sperm-specific counters produced, one being the Makler counting chamber I believe.

[Microfan]

I've read other comments on how the thickness of the coverslip is crucial, but I don't understand. isn't the volume of the sample under the coverslip already determined by the distance between the under surface of the coverslip and the depth of the chamber? If the coverslip rests on the posts of the counting chamber then how does the thickness of the coverslip have a bearing on the volume of liquid under it? is it that thinner coverslips are lighter and may float on the sample rather than sit upon it? Thanks for any additional info.
I looked up the Makler counting chamber and it is a bit over my price range of $100, in fact the ones I found were ~ $700-1000 Nice piece of equipment though!!

[dtsh]

I've read other comments on how the thickness of the coverslip is crucial, but I don't understand. isn't the volume of the sample under the coverslip already determined by the distance between the under surface of the coverslip and the depth of the chamber? If the coverslip rests on the posts of the counting chamber then how does the thickness of the coverslip have a bearing on the volume of liquid under it? is it that thinner coverslips are lighter and may float on the sample rather than sit upon it? Thanks for any additional info.
I looked up the Makler counting chamber and it is a bit over my price range of $100, in fact the ones I found were ~ $700-1000 Nice piece of equipment though!!

I agree with you that I don't think the coverslip thickness is a requirement as the volume under measure is below the coverslip, and therefore thickness of the coverslip doesn't matter. That said, I have heard it claimed that a #1.5 coverslip will flex, changing the volume, but I don't know that there's any validity to it and don't have the precision to measure such a difference.

The proper 0.4mm coverslips are nicer to work with and are more durable.

[Microfan]

I've read other comments on how the thickness of the coverslip is crucial, but I don't understand. isn't the volume of the sample under the coverslip already determined by the distance between the under surface of the coverslip and the depth of the chamber? If the coverslip rests on the posts of the counting chamber then how does the thickness of the coverslip have a bearing on the volume of liquid under it? is it that thinner coverslips are lighter and may float on the sample rather than sit upon it? Thanks for any additional info.
I looked up the Makler counting chamber and it is a bit over my price range of $100, in fact the ones I found were ~ $700-1000 Nice piece of equipment though!!

I agree with you that I don't think the coverslip thickness is a requirement as the volume under measure is below the coverslip, and therefore thickness of the coverslip doesn't matter. That said, I have heard it claimed that a #1.5 coverslip will flex, changing the volume, but I don't know that there's any validity to it and don't have the precision to measure such a difference.

The proper 0.4mm coverslips are nicer to work with and are more durable.

Thanks for your reply, I will look for the recommended 0.4mm coverslips.

[Tom Jones]

Pasteur pipettes are a little difficult to use with such small volumes. Capillary tubes are better for loading the chambers. Cheaper, too.

The thicker cover slips are heavier and less prone to float on top of the specimen if it's viscous. Otherwise, either one works fine. Unless you get one with the chamber, you may want to stick to the regular ones. Replacement coverslips for counting chambers are available, but cheap they're not:

https://www.sigmaaldrich.com/US/en/prod ... h/br723015
https://www.amazon.com/Hemacytometer-Co ... B008R42WV6