System for in situ reactivation of fluorescence marker

摘要

Vapor is provided locally at a sample surface to allow fluorescence of the fluorescent markers in a vacuum chamber. For example, a nanocapillary can dispense a liquid near a region of interest, the liquid evaporating to increase the vapor pressure near the fluorescent markers. The increase in vapor pressure at the fluorescent marker is preferably sufficiently great to prevent deactivation or to reactivate the fluorescent marker, while the overall pressure in the vacuum chamber is preferably sufficiently low to permit charged particle beam operation with little or no additional evacuation pumping.

主权项

1. A system for analyzing a specimen, comprising:
a vacuum chamber; an electron beam column for scanning a focused electron beam over a surface of a sample in the vacuum chamber; an electron detector for detecting secondary electrons emitted from the sample in response to an impact of the electron beam to form an image of the sample; an optical system for illuminating the sample within the vacuum chamber with a wavelength of light that causes fluorescence of fluorescent markers in the sample and for detecting fluorescence from the fluorescent markers; and a nanocapillary that dispenses a liquid that evaporates, the nanocapillary configured for locally applying vapor to the surface of the sample at pressures sufficient to increase an activity of the fluorescent markers while maintaining a lower background pressure in the vacuum chamber.