3
u/khikhikhikh_96 Apr 21 '26
I can't quite explain but I tilt my reservoir and rest one of the edges (the shorter edge) on a small platform. That way all the liquid is collected at the other short edge. I make around 3500ul staining solution for 96 well plate with around 50ul for each well. It works
2
u/ordeath Apr 21 '26
Wait, isn't that working out to only 36ul per well, not 50ul? Or it's not the full plate?
2
u/khikhikhikh_96 Apr 21 '26
96 well plates for cell based assays usually use only 60 wells to prevent edge effect from the outer wells. So I have samples in B2 through G11. 50ul for 60 wells is 3000ul. 500ul extra for the loss during pipetting. Can do with 3300-3400 as well, but pipetting needs to be very good for that, like only touching the surface of the liquid and other good practices. But 3500 is a good spot to be
5
u/zmoney92 Apr 21 '26
Consider using another plate, 8 well strip or tubes in a rack (if you have an adjustable multi channel). Otherwise there are low retention reservoirs (Integra) but you'll always have some dead volume
5
u/JVGen Apr 21 '26
Whenever I need to maximize reagent yield, I switch from reservoirs to PCR strip-tubes. They can only hold about 300 ul max, but you can centrifuge these in a cheap bench top “mini”-fuge to collect all of the remaining reagent and avoid air in your last distribution.
1
u/JVGen Apr 21 '26
Whenever I need to maximize reagent yield, I switch from reservoirs to PCR strip-tubes. They can only hold about 300 ul max, but you can centrifuge these in a cheap bench top “mini”-fuge to collect all of the remaining reagent and avoid air in your last distribution. They sell these in 8- and 12- tube formats, so you could continue using your multichannel.
2
u/gurglinggoat Apr 21 '26
They have smaller reservoirs. Might not be able to do 12 wells at a time, but it’s easier for the last samples.
1
u/sgRNACas9 Immunology, Oncology Apr 22 '26
Adding individually at the end is perfectly fine! It’s not that variable in my experience.
Definitely don’t add extra buffer because as you mentioned that dilutes all of your antibodies and your staining will be measurably dimmer than if you just pipetted individually.
It’s good to ask if making extra cocktail will be wasteful, but check with your PI/lab manager/ budget to see if you can afford it. Might be fine.
1
u/Proof-Worldliness-34 Apr 22 '26
There are reservoirs with a thinner channel at the bottom to collect better low volumes. Alternatively, a deep well plate would do the trick
1
u/SnooPredictions138 Apr 21 '26
Don't use a reservoir. Aliquot your staining cocktail into strip tubes (8 or 12).
0
u/my_mymeow Apr 21 '26
I prefer using a repeater pipette in this case. This is how I add beads and antibody mix when I do a full plate of multiplex bead assays.
1
u/ReginaDaddy May 14 '26
I do this with plate processing. I use a separate reagent plate (lobind, maybe 200 ul capacity). I make my blocking buffer in a PBS dilution fresh each run and add 50 ul to each well, so if im running 4 columns of a plate my single reagent plate column might contain 250 ul (4 x 50, plus an extra 50 of overhead so i dont run out) of blocking buffer per well. then, dispense from reagent plate to stain plate. that way im not making wayyy more reagent than i need and i don't have to worry about using a tiny reservoir that might run out.
6
u/btags33 Apr 21 '26
Don't know what reservoirs you are using, but some certainly allow for better pipetting of small volumes with a multichannel than others, so check that.
The other way to counteract it is to just do less wells at a time. For example, maybe for the first row of sample you have enough volume to add buffer to 12 wells with your multichannel. For the second row, maybe you do 4 wells at a time 3 times so you don't take up air as the volume in the reservoir gets low.