Counting Cells

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In this video we will show you how to count cells using a hemocytometer. This protocol begins after our cells have been trypsinized and collected. To learn more about this and tissue culture in general, please see our video on Getting Started with Tissue Culture.
We’ve broken this video up into two major components for ease of use; the laboratory work involving setting up the hemocytometer and counting the cells, followed by the calculations you’ll need to determine your cell density.
______________________________________________________________________________________________
0:00 Intro
0:41 Materials
0:58 Laboratory Protocol
1:41 Counting Cells
3:18 Calculating Viable Cells
3:56 Cleanup

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Опубликовано:

5 авг 2024

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Комментарии : 17

@addgene Год назад

Addgene does not regularly monitor comments posted here, so we may not see your question immediately. We’d be happy to answer any questions sent to help@addgene.org as soon as possible. Please include the name of the video along with any questions so our support team can help. Thanks!

@Koonnuusana 3 года назад

Thank you so much for this good video. It make me clearly understand about how to counting cell in cell culture. 😊❤🍀👩‍🔬

@exotoxinslab Год назад

I love the video. I would like to propose an improvement. For many people, this will be the first time in contact with a counting chamber. It will be useful to say what kind of chamber it is and what type of cells you are counting. The Neubauer chamber (shown here) has two different sections for counting: One for erythrocytes and platelets (the one shown in this demo , I think?) and another section for lymphocytes (which are bigger). To decide which one to use, you need to know what size of cells you are expecting. And depending on the section, you will have a different multiplication factor. This will be helpful for people doing research on other types of cells, like fungi or protozoa. The explanation in ru-vid.com/video/%D0%B2%D0%B8%D0%B4%D0%B5%D0%BE-rR1ov4VEJXQ.html allows to think about the reason for the calculations. 🙂

@nurulhusnahassan1223 3 года назад

how can we calculate the number of cell when using only normal microscope slide and coverslip (not hemocytometer) ? I am currently working on stomach analysis, in wish I observe the stomach content under compound microscope.

@batoulmed6449 3 года назад

this is very helpful! thank you

@binayakdas7361 4 года назад

Why is tryphan blue unable to penetrate the viable cells but nonviable ones?

@addgene 4 года назад

Thanks for the question. This is because trypan blue is not membrane permeable. Viable cells have an intact membrane, which is why trypan blue won’t cross the membrane to stain them.

@irishguy200007 3 года назад

Can software count cells without grid lines?

@addgene 3 года назад

Some software may be able to count the cells in a field of view. The gridlines are important for determining the boundaries of the chamber, helping you to calculate an accurate concentration.

@funny11744 2 года назад

Please mention how to count and distinguish ADSC ( Adipos derived stem cell) from other cells from SVF , using HEMOCYTOMETER and microscope. What îs the TOTAL magnification( eye piece and lens of microscope) îs required ? Thanks

@addgene 2 года назад

Thank you for your question. This video is meant to provide a general procedure for counting cells with a hemocytometer. Addgene does not perform experiments with the cell types you mentioned, so we can’t say for certain how to best distinguish between them or what the required magnification might be for your experimental context. In general, a hemocytometer is not intended to be used with a high magnification lens.

@islandsoul679 2 года назад

What microscope and screen is that??

@funny11744 3 года назад

Please answer to this question : why solution factor îs 2 ?

@maturtlelaza 3 года назад

Dilution Factor is just a number correcting for any added volume. Because equal parts of cell culture and trypan blue were combined, it means that the original cell culture will be TWICE as concentrated as the tested solution.

@addgene 3 года назад

The dilution factor in our video is 2, because we diluted our sample 1:1 in trypan blue. Your dilution factor may change depending upon how much trypan blue you add.

@irishguy200007 3 года назад

Why do I need a hemocytometer if I Am using software

@addgene 3 года назад

Software may be great for doing the actual counting and obtaining the number of cells in a field of view; but, a hemocytometer creates an environment of known size so that the cell count can be translated into a concentration.