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Using Multiphoton Microscopy to See Chromosomes in Action

ecimen, MPM delivers crisp, clear images, even in thicker tissue samples like Drosophila salivary glands.

The research was ultimately possible thanks to the unique composition of the fruit flies' so-called polytene cells, giant, multistranded chromosomes with hundreds of sets of the genome instead of the usual two sets in conventional cells. This enlarges the usual nuclear dimensions by about 10 times, making them large enough to image the detail.

The results were stunning. "Within two weeks we had spectacular pictures," said Lis. The images included pictures of the genes (hsp70 genes) that protect flies from the effects of extreme heat. By cranking up the heat, the researchers could activate these genes, and by using fruit flies specifically bred to carry fluorescent proteins on HSF, they could watch the transcription factors in action.

"This is the first time ever that anyone has been able to see in detail, at native genes in vivo, how a transcription factor is turned on, and how it then is activated," said Webb.

Using another method that Webb engineered at Cornell, called fluorescence recovery after photobleaching, the researchers also discovered that HSF activators bind to hsp70 genes much longer than previously thought before being replaced with new HSFs, which raises new questions about the mechanisms of gene transcription.

The technique also may offer a new tool for researchers across the biological sciences. Webb says it marks the success of an interdisciplinary trend that offers new potential for researchers in a variety of fields.

"Interaction between the physical sciences and the life sciences is very powerful," said Webb. "And it's becoming more powerful as a tool for advancing our understanding of the life sciences."

Better understanding transcription in lower organisms will help understand the processes in higher organisms, Yao added. "We hope to push the limits to huma
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