Protocols

With the right preparation and a clear plan of action, useful (and beautiful!) cell images can be yours. Find icon-based, step-by-step instructions for common imaging protocols, including tips for a successful experiment, advice on controls, and lists of needed supplies.

Fix, perm, & block

General instructions for fixation, permeabilization, and block to prepare your cells for immunolabeling. Fixing and permeabilizing cells generally locks them in place and makes it possible for larger molecules such as antibodies to access the interior of the cell.

Immunolabeling using secondary antibodies

Immunolabeling is a technique for fluorescently labeling a specific biological target within a sample using an antibody. This protocol provides general instructions for immunolabeling cells with a primary and secondary antibody.

Nuclear staining

For most nucleic acid stains the fluorescent signal is minimal before binding to nucleic acids, and there is a significant increase in fluorescence intensity after the dye has bound the nucleic acid. Find general instructions for labeling the nuclei of cells using permeant or non-permeant nucleic acid dyes.

F-actin staining

When phalloidin, a molecule that binds F-actin, is conjugated to a fluorophore, it becomes a very useful tool for cell biologists who are using fluorescence microscopy to study their cells. Find general instructions for labeling F-actin in fixed and permeabilized cells.

Functional mitochondrial staining

Tetramethylrhodamine, methyl ester (TMRM) is a cell-permeant dye that accumulates in active mitochondria with intact membrane potentials. This protocol provides basic instructions to perform a fluorescence-based assay to detect functional mitochondria.

Mounting coverslips

Formulations of mounting media that can add favorable properties such as optimizing the refractive index to match that of glass, preventing photobleaching, or preserving samples for long-term storage are widely available. Find general instructions for mounting media on coverslips.