Mix thoroughly and then begin to do serial dilutions with saline until you get to a point that you can count cells. You may need to use a DNA stain and a hemocytometer to do this accurately. Replicate each sample a few time to ensure that the process is repeatable.

Mix thoroughly and then begin to do serial dilutions with saline until you get to a point that you can count cells. You may need to use a DNA stain and a hemocytometer to do this accurately. Replicate each sample a few time to ensure that the process is repeatable.

thanks Britt... it will be highly appreciable if you upload scientific literature or give link...

Here is link for one procedure: http://www.abcam.com/protocols/counting-cells-using-a-haemocytometer 

Here is another link and two downloaded files for instructions. I hope this will help you.

Raj,

question is more what cells are you interested in? The composition of colostrum is VERY different to normal milk, and remember, SSC counts just cells. Whats your actual aim?

hello Werling.. you are very right,, colostrum is different from milk. my interest is in all types of  somatic cells in colostrum. Britt... thanks for the link... but I think, counting cells in colostrum is somewhat different from that of in blood. have you got any experience or literature which indicated use of haemocytometer for colostrum

ATTACHED is a paper from 1963, which is the only one that I could find that is available online. You will need to dilute the colostrum, but this paper should provide methods for staining and observing cells. ALSO, you might try adding some EDTA to the diluted samples to prevent clumping and binding.

Britt.. thanks for your efforts... I am following similar procedure for colostrum... but facing problem as mentioned above...

Did you try to wash cells before counting them ?

For youre you have to dilute your colostrum first (5 fold?) Once you've diluted your colostrum you can gently centrifuge it  (let's say 400 to 700xg, 20 min, 4 to 20°C) to precipitate cells, discard the fat and protein rich supernatant and resuspend clean cells in an equal volume of buffered saline... that should get rid of most of the interferences for subsequent cell staining and counting... what do you think?

hello Bianchi,, i am not diluting but directly centrifuging the sample. let me try your suggestion.

I tried another method which gave good results. 1st prepare smear of colostrum (1x1 cm sq), dry it and dip in xylene for 2-3 minutes. then stain with newman-lampert stain and observe under oil immersion. you will able to count cells.

I am not sure fo the level of precision that you wish to obtain. For an estimate and an evaluation of the cow status (inflammation, no inflammation), the CMT (California Mastitis Test).is easy to use will give you a fair estimate. We often forget that CMT has a ph indicator. It will turn purple when ph is alcaline (yellow when acidic). Milk and colostrum will be alkaline in presence of inflammation, du to higher bicarbonate level in the milk. Density and appearance of the mixture will also change with higher concentration of cells.  You have to use it repeatedly to refine interpretation. Comparing with lab results as described previously will help. 

I would suggest to use a DCC counter from Delaval and then make serial dilutions using milk with low SCC. Saline buffer, water or whatever seems to interfere with the lecture of the DCC counter

@ Hey Bouchard,,,, no doutd cmt is good for evaluating udder health status,, but i want to estimate scc for my work.

@ Hello castellano... I tried using delaval scc counter by diluting colostrum but ends with clogging of tubes. i read one of your paper on goats, in which you estimated scc in colostrum of goat upto 10hr postpartum. have you tried this in cow colostrum. Also, can you send me the detailed step by step procedure for estimating scc in goat colostrum using delaval machine

What kind of diluent did you use? It should work if you make serial dilutions with your colostrum and low SCC cow milk. SCC are specially high in goats and goat colostrum is really dense. Therefore, if you measure your cow colostrum using this procedure, it should work

I used DW upto 50: 50 ratio. how much dilution did you made?