In Part I, cell types for brightfield-based cell counter was introduced. To review, click https://www.blog-nanoentek.com/post/ch-5-1-types-of-cell-used-for-automated-cell-counter-part-i.
As the principle of cell counting is different for counters based on brightfield and fluorescence, optimal cell types for quantification also differ.
To compare with brightfield-based,
a fluorescence-based automated cell counter uses dye that only stains the nucleus of the cell, which means that only nucleus containing cells are stained for quantification. It is, therefore, considered more accurate because it excludes any unwanted substances other than cells from being counted.
[Fluorescence sample]
For fluorescence-based, most of cells with nucleus can be used for quantification. Not only cell lines but also primary cells can be used regardless of the existance of debris or waste.
So what are primary cells?
Primary cells are cells that have been obtained directly from living organism tissue. Two different types of primary cells - adherent and suspension - are present. While both are used for many research, adherent cells are more specifically used for cytology and suspension cells are used for bulk protein production.
Adherent cells | Suspension cells |
 Cells that grow attached to the ground |  Cells that are floating in the medium |
Most commonly used primary cells are as follows:
- Epithelial cells | - Endothelial cells |
- Fibroblasts | - Muscle cells |
- Keratinocytes | - Hematopoietic cells |
- Melanocytes | - Mesenchymal stem cells |
Using primary cells have advantages & disadvantages:
Advantages | Disadvantages |
- Similar to in vivo environment thus has high relevance - Allows to perform less animal testings | - Expensive - Limited proliferation - Take longer time to grow compare to cell lines |
To summarize, a brightfield-based counter quantifies cells by staining only the dead cells (cytoplasm area including nucleus) whereas a fluorescence-based counter quantifies cells by staining only the nucleus of cells. Depending on what you need for your research and cell sample, two different types of cell counter are your options.
Check out NanoEntek cell counters below.
NET products :
Fluorescence-based cell counters
ADAM MC2 & CellT
Analysis time | < 25 seconds |
Dye | Acridine orange (AO) / Propidium Iodide (PI) |
Cell type | Cell line, PBMCs, adipose stem cell, primary cells etc. |
Measuring range | 5x10E4 to 2x10E7 |
Brightfield-based cell counters
An automated cell counter
Analysis time | < 20 seconds |
Dye | Trypan blue |
Cell type | Cell lines |
Measuring range | 1x10E4 to 1x10E7 cells/mL |
The world's fastest automated cell counter
Analysis time | < 1 second (manual focus) < 10 seconds (auto focus) |
Dye | Trypan blue |
Cell type | Cell lines |
Measuring range | 1x10E4 to 2x10E7 cells/mL |
A high-throughput automated cell counter
Analysis time | 3 minutes / 48 tests |
Dye | Trypan blue |
Cell size range | Detectable : 1 - 85 μm Optimal : 5 - 80 μm |
Measuring range | Detectable : 1x10E4 to 2x10E7 cells/mL Optimal : 1x10E5 to 1x10E7 cells/mL |
References:
1. “Adherent Cell Culture vs. Suspension Cell Culture.” Thermo Fisher Scientific - US, https://www.thermofisher.com/kr/ko/home/references/gibco-cell-culture-basics/cell-lines/adherent-vs-suspension-culture.html.
2. Primary Cells | ATCC. https://www.atcc.org/cell-products/primary-cells.
3. Briggs, James. “Benefits of Using Cell Lines versus Primary Cells.” VH Bio, 15 Jan. 2021, https://www.vhbio.com/benefits-using-cell-lines-versus-primary-cells/.
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