Abstract: The invention provides a method of determining a measure of a density of markers in a sample, and an apparatus arranged for performing said method. In particular said method comprising the steps of: irradiating an illumination region of the sample with light, wherein the markers present in the illumination region of the sample emit fluorescence light in response to the irradiation with light, detecting an intensity of the fluorescence light from a detection region of the sample, comprising at least a part of said the illumination region, irradiating an area within said detection region of the sample with a focused charged particle beam to deposit a dose of charged particles in said area, and determining a measure of the density of markers in said area using a change of the detected intensity of the fluorescence light as a function of the deposited dose of charged particles in said area.

Abstract: The invention relates to a method for inspecting a sample with an assembly comprising a scanning electron microscope (SEM) and a light microscope (LM). The assembly comprises a sample holder for holding the sample. The sample holder is arranged for inspecting the sample with both the SEM and the LM, preferably at the same time. The method comprising the steps of: capturing a LM image of the sample in its position for imaging with the SEM; determining a position and dimensions of a region of interest in or on the sample using the LM image; determining values to which the SEM parameters need to be set to image the sample at a desired resolution; and capturing a SEM image of the region of interest, preferably using the first electron beam exposure of said region of interest.

Abstract: An apparatus for inspecting a sample, is equipped with a charged particle column for producing a focused beam of charged particles to observe or modify the sample, and an optical microscope to observe a region of interest on the sample as is observed by the charged particle beam or vice versa. The apparatus is accommodated with a processing unit adapted and equipped to represent an image as generated with the column and an image as generated with the microscope. The unit is further adapted to perform an alignment procedure mutually correlating a region of interest in one of the images, wherein the alignment procedure involves detecting a change in the optical image as caused by the charged particle beam.

Abstract: The present invention relates to a method for mutually aligning a scanning electron microscope SEM and a light microscope LM by creating a change (61) in the detected light signal of the light microscope LM by illuminating a substrate with an electron beam, correlating the position of the electron beam in the coordinate system of the scanning electron microscope SEM to the position of the observed change in the detected light signal in the coordinate system of the light microscope LM, and relatively shifting the scanning electron microscope SEM and the light microscope LM with respect to one another to a desired relative position of the coordinate systems (60, 62).

Abstract: An inspection apparatus is provided comprising in combination at least an optical microscope (2, 3, 4) and an ion- or electron microscope (7, 8) equipped with a source (7) for emitting a primary beam (9) of radiation to a sample (10) in a sample holder. The apparatus may comprise a detector (8) for detection of secondary radiation (11) backscattered from the sample and induced by the primary beam. The optical microscope is equipped with an light collecting device (2) to receive in use luminescence light (12) emitted by the sample and to focus it on a photon-detector (4).

Abstract: An inspection apparatus is provided comprising in combination at least an optical microscope (2, 3, 4) and an ion- or electron microscope (7, 8) equipped with a source (7) for emitting a primary beam (9) of radiation to a sample (10) in a sample holder. The apparatus may comprise a detector (8) for detection of secondary radiation (11) backscattered from the sample and induced by the primary beam. The optical microscope is equipped with an light collecting device (2) to receive in use luminescence light (12) emitted by the sample and to focus it on a photon-detector (4).