The establishment of rat basophilic leukemia (RBL) cell lines in the 1970s offered researchers a convenient, if imperfect, model system for the study of difficult-to-obtain mast cells. RBL-2H3 cells subcloned from one of the original RBL cell lines have been used to study the biochemical pathways for secretion in mast cells. Some RBL cell lines are grown as suspension cultures and others are adherent. Because the morphology of the cells under different growth conditions, or even within the same culture, can vary, the ability to create a customized StainFree analysis setting is extremely useful in ensuring accurate cell counts.!
Since there are often some adherent cells present in suspension cultures of RBL cells, a user-created analysis setting in SoftMax Pro Software can be very helpful in ‘training’ the software to identify only the cells with suspension-type morphology and exclude the attached cells. For adherent cultures, which may vary in their morphology, a user-created analysis setting maximizes identification of all cells regardless of shape.
RBL 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||Setting for cell counts|
|Optical configuration||SpectraMax MiniMax 300 Imaging Cytometer|
|Wavelength settings||Transmitted light|
|Image acquisition settings||Exposure: 7 ms
Focus adjustment: 20 µm
|Image analysis settings||Analysis type: Discrete Object Analysis
Wavelength for finding objects: TL
|Find objects||Settings: Create new setting|
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.