Application Note

Improved cell counting method using transmitted light on the SpectraMax MiniMax cytometer

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Hematoxylin, a natural product extracted from the heartwood of the logwood tree (Haematoxylin campechianum), is the most commonly used dye for staining protocols in histology. Hematoxylin reacts like a basic dye with a purplish-blue color and stains acidic or basophilic structures, including the cell nucleus and organelles that contain RNA, such as ribosomes and the rough endoplasmic reticulum1.
The SpectraMax® i3x Multi-Mode Microplate Reader with the SpectraMax® MiniMax™ 300 Imaging Cytometer is equipped with transmitted light (TL), green fluorescence (Ex/Em: 460/541), and red (625/713) fluorescence channels. The MiniMax cytometer with SoftMax® Pro Software can be used to image and analyze a variety of cells. Here, we demonstrate how hematoxylin can be used to increase the contrast in transmitted light images to improve cell counting for cell lines with relatively flat morphology or indistinguishable intercellular space.

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