Along with new cutting-edge methods, Cold Spring Harbor Protocols is home to an in-depth library of basic laboratory methods. The just-released April issue features articles that exemplify the attention to lab standard techniques.

Dye Loading with Patch Pipettes from Arthur Konnerth and colleagues from the Institut fur Physiologie der Ludwig-Maximilians-Universitat Munchen, describes the loading of individual cells with fluorescent probes via patch pipettes. This method allows for combined electrophysiological and optical measurements at a quantitative level. The patch-clamp methodology has been successful for single-cell dye labeling in cultured neurons, brain slices, and in vivo preparations. A wide range of dyes can be loaded using this method, including probes for morphological reconstruction, ion-sensitive indicator dyes for monitoring second-messenger cascades, and dye-labeled proteins for fluorescence resonance energy transfer (FRET), fluorescence correlation spectroscopy (FCS), and fluorescence recovery after photobleaching (FRAP) studies. The most widespread application of this technique has been for Ca2+ imaging.

Like all our featured articles, Dye Loading with Patch Pipettes is freely available to subscribers and non-subscribers alike.