First cultured by Greene and Tischler in 1976, PC-12 cells originated from a pheochromocytoma (neuroendocrine tumor) of the rat adrenal medulla. It was developed as a model cell line and an alternative to adrenal chromaffin primary cell cultures. PC-12 cells are able to differentiate into neuron-like cells in the presence of nerve growth factor or dexamethasone. Due to their differentiation ability and ease of culture, PC-12 cells are used in a variety of research areas ranging from drug efficacy to neurosecretion.
To count PC-12 cells without staining, we recommend creating a new setting using the drawing tools in the software. Choose the Discrete Object Analysis and then use the drawing tools to define cells and non-cell areas in your max and min images. To get more accurate counts in dense clusters of cells, try drawing just within the borders of the cells. Conversely, if you find that the software is over counting the cells in your images, try drawing slightly outside the borders of the cells.
It’s easy to get growth curves with the Field Analysis feature in SoftMax Pro Software. Field Analysis calculates the percent area covered by cells (confluence) in your images. Measuring cell confluence can be a valuable tool for better assay development. For example, you may seed cells at two or more test densities, calculate the confluence of your cells just prior to assay, and upon obtaining assay results, determine what level of confluence yielded the best results.
PC-12 Cells Analysis Toolkit
- SpectraMax® i3 Multi-Mode Microplate Detection Platform
- SpectraMax® MiniMax™ 300 Imaging Cytometer
- SoftMax® Pro Software
|Parameter||Setting for cell counts||Setting for growth curves|
|Optical configuration||SpectraMax MiniMax 300 Imaging Cytometer|
|Wavelength settings||Transmitted light|
|Image acquisition settings||Exposure: 5 ms||Exposure: 7 ms|
|Image analysis settings||Analysis type: Discrete Object Analysis Wavelength for finding objects: TL||Analysis type: Field Analysis Wavelength for finding objects: TL|
|Find objects||Predefined setting: CellsA|
|For PC-12 cultures, a user-defined setting may give the best results. Image acquisition and analysis settings may vary depending on assay conditions, microplate used, and other factors.|
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.