Practical advice and lab tips from Biotium scientists on using fluorescence in your research
Tech tip: Imaging Bacteria Using Agarose Pads
Bacteria can be more difficult to image than other cell types because they are small, usually non-adherent, and are often motile. Here we describe how to image live or fixed bacteria using agarose pads to immobilize the cells during the imaging process, allowing you to perform multi-color fluorescence microscopy experiments.
Tech Tip: Battling Tissue Autofluorescence
Autofluorescence in tissues from lipofuscin, red blood cells, and extracellular matrix can make it all but impossible to distinguish specific fluorescence staining from non-specific background. See our protocols for quenching autofluorescence with TrueBlack®.
Tech tip: Avoiding Artifacts from UV Photoconversion of DAPI and Hoechst
In multi-color imaging experiments, limiting cross-talk between detection channels is crucial for obtaining specific results. Many researchers are unaware that photoconversion of DAPI and Hoechst can cause nuclear stains to bleed into the FITC channel. Learn about the different ways to avoid it.
Tech Tip: Combined Direct and Indirect Immunofluorescence Using Primary Antibodies from the Same Host
Multicolor staining using fluorescent secondary antibodies provides signal amplification for greater sensitivity, but requires that primary antibodies be raised in different hosts. Directly labeled primaries from the same host can used simultaneously, but signal can suffer. A combination of indirect and direct methods takes advantage of the best of both for added flexibility in multicolor staining. See our protocol to learn more.