Derived from the blood of an acute monocytic leukemia patient over 30 years ago, the human monocytic cell line THP-1 is widely used to study the function of monocytes and macrophages, as well as in leukemia research. Normally THP-1 cells grow in suspension, but they can be induced to differentiate into adherent macrophage-like cells, which express IL-8 and other markers, and can thus serve as a model system for differentiation.
Figure 5: IL-8 ELISA
For suspension cells, such as undifferentiated THP-1 cells, you can use a lower exposure time to minimize blurring caused by the floating motion of the cells. The SpectraMax MiniMax 300 Imaging Cytometer is able to switch wavelengths very rapidly, minimizing the amount of cell movement that can occur between images and leading to better overlay of images taken in different channels. The ‘CellsC’ predefined TL image analysis setting works well for suspension cells with rounded morphology, while ‘CellsA’ more accurately identifies the differentiated, more flattened cells.
THP-1 Cells Analysis Toolkit
- SpectraMax® i3 Multi-Mode Microplate Detection Platform
- SpectraMax® MiniMax™ 300 Imaging Cytometer
- SoftMax® Pro Software
Instrument Settings for imaging and cell counting
|Parameter||Undifferentiated THP-1||Differentiated THP-1|
|Optical configuration||SpectraMax MiniMax 300 Imaging Cytometer|
|Wavelength settings||Transmitted light, 713|
|Image acquisition settings||Exposure: 2 ms (TL), 17 ms (713)||Exposure: 6 ms (TL)|
|Image analysis settings||Analysis Type: Discrete Object Analysis
Wavelength for Finding Objects: TL or 713
|Find objects||Predefined setting: CellsC||Predefined setting: CellsA|
About StainFree Cell Detection Technology
Imaging cell-based assays typically requires the use of fluorescent probes that can be toxic to living cells or may only function in fixed cells. A label-free method for analyzing cell counts and cell confluence enables researchers to quantitatively monitor cell proliferation and health without time-consuming workflows that may disrupt cell viability.
The SpectraMax i3 Multi-Mode Microplate Platform with MiniMax 300 Imaging Cytometer uses unique, patent-pending StainFree Cell Detection Technology that allows you to perform cell proliferation, cytotoxicity, and other assays without nuclear stains like DAPI, which intercalates with DNA, or live cell dyes that are actually toxic to cells in the long term.