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 method of processing a work piece, comprising:
a. placing a work piece in a vacuum chamber; b. evacuating the vacuum chamber; c. exposing a new surface of the work piece; d. illuminating fluorescent markers in the new surface while maintaining a background vacuum pressure in the vacuum chamber by locally providing vapor at the new surface, wherein the background vacuum pressure is lower than a local vapor pressure at the new surface; e. acquiring an optical image of the fluorescent markers in the new surface; and f. repeating steps b through e.