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Chapter 3
– Protocol Setting
The LUNA-II
™ Automated Cell Counter provides a basic (DEFAULT) protocol which can be used for
most common cell lines. In addition, LUNA-
II™ also provides tools to setup individual protocols. All
the parameters in the protocol can be modified by users and saved as alternative protocol. Users
can save up to 300 protocols. These functions provide personalized protocol for optimal use.
3.1 Parameters in the Protocol
In the protocol menu, the LUNA-II
™ Automated Cell Counter provides following parameters
Parameter Range DEFAULT
Dilution Factor 1
‒ 100
2
Noise Reduction 0
‒ 10
5
Roundness 0
‒ 100%
60
Min. Cell Size 3
‒ 59 µm
3
Max. Cell Size
4 ‒ 60 µm
60
Declustering Level None, Low, Medium, High Medium
Dilution Factor: The value for dilution factor in the DEFAULT protocol is preset as 2 for With
Trypan Blue. However, users can modify this value according to the dilution of the original sample.
The Dilution Factor can be adjusted either by a scale of 1 or a scale of 10 between 2 to 10 and 10-
100, respectively. The Dilution Factor is used to automatically calculate the concentration of cells in
original sample from the cell counting result. Adjusting Dilution Factor will be helpful for users
handling high density cells such as fermented CHO cells. In such cases, serial dilutions and
repeated counting will be needed with appropriate Dilution Factor.
Noise Reduction: Noise Reduction means the decrease of the background for counting. With
higher Noise Reduction, the instrument dose not detect faint signals of weakly stained objects. With
lower Noise Reduction, the instrument can detect objects with faint signals. Since staining intensity
of cells with trypan blue may vary from cell to cell, adjusting Noise Reduction will be helpful for
optimal detection of specific type of cells.
Roundness: Roundness refers to the roundness of the objects in the image. Since the shape of
cells may vary and not complete sphere, adjusting Roundness enables optimal detection of cells.
The instrument with higher Roundness counts objects with more roundness as cells and excludes
objects with less roundness for counting as cells. LUNA-
II™ with lower Roundness will be suitable
to count cells with irregular shape since it detects objects with less roundness as cells.