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Counting bacteria

Counting bacteria - Microbiology Forum

Counting bacteria - Discuss Microbiology Science and Protocols here. Post questions on the study of viruses, fungi, parasites and bacteria here. Microbiology Forum.


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  #1  
Old 10-18-2008, 03:54 PM
RUTH SIELE
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Default Counting bacteria



Hi all,

Thanks in advance for your help.

I have bacteria and a membrane

My objective is to analyze the deposition of the bacteria on the membrane with time.

To do this, - I have prepared 0.1 OD (600) bacterial solution
***************- I run the solution through the membrane in a flow cell under flourescent
**************** microscope
************** - I took pictures of the deposited bacteria in every 15 minutes using 40x
**************** objective lens.
*Now, i want to Aanlyze the number of bacteria deposited on the membrane with time.

* Thus, instead of counting all the deposited bacteria one by one, I am looking for any*
** othere means which can help me get the number of deposited bacteria per unit area of*
** the membrane? I was using the 40x lens.

Thanks so much again for your help,

looking forward to hearing from you
*
Sincerly yours,
*
Ruth

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  #2  
Old 01-20-2009, 05:45 AM
Pipette Filler
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Default Re: Counting bacteria

You want to use sampling. Counting every bacteria is a lot of work. Decide on how many digits of precision you require in your answer and start to plan a sampling stategy with that level of precision in mind.

The idea is to sample a fraction of the membrane. The precision is not related to the fraction of the area of the membrane you use in counting, but in the number of counts that you make. Think of it this way.

Suppose you wanted to know how many letter 'e's were in a book. There are lots of 'e's in many languages. You quickly get a large number using a few pages. Determining the number of 'x's is trickier. The letter is rarer. You look over many more pages to learn about the number of 'x's.

There is an unbiased counting method called the counting frame. This device and the sampling method called the fractionator can be combined to estimate the density of objects in an unbiased manner.

See the book by Howard and Reed for a description of the methods.
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