New from Andor: Microspectroscopy using dynamic slit

Microscopy images show objects magnified on a scale down to the refraction limit of light.

Depending on the light source and filters in the beam path, certain properties of a sample can be visualized. The underlying photophysical processes may differ. The light distribution and contrast can be generated by absorption, reflexion, fluorescence emission, scattering, phase contrast or a Raman process.

Besides the varying intensity, each point contains spectral information. This fact is, for example, utilized in hyperspectral imaging where data is recorded as a multidimensional array.

There are several methods to gain spectral information from a microscopy image. Microscopes offer the option to attach an optical fibre to one of the output ports. The light coupled into the fibre can be spectrally resolved by a spectragraph. The disadvantage of this method is that two detectors are necessary, one camera for the image acquisition and another one which is connected to the spectrograph. Andor now offers an add-on solution to capture both, an image of the sample and a local spectrum, with the same CCD detector. The dynamic slit replaces the standard input slit on the Shamrock SR-303i. It can be opened to a width of 12 mm.

Fig. 1: Typical setup of a Shamrock SR-303i connected to the side port of a microscope.

The image can be relayed on the CCD sensor by moving the grating into 0th order position. The CCD is read out in image mode and the acquired image can be viewed in the software as usual. When the sample is moved and centered around the region of interest, the slit is narrowed to an appropriate width (Fig. 2). Then the grating is returned into the position of 1st order reflection. Taking the next acquisition gives the spectrum from the chosen region of interest. On the vertical axis, each pixel represents a different spectrum. With vertical binning enabled, the intensity from more than one row can be combined to give a better signal-to-noise ratio.

The dynamic slit is an interesting accessory for applications like

Raman microspectroscopy

Confocal Raman microspectroscopy

Fluorescence microspectroscopy

Multi-photon microspectroscopy

Micro photoluminesce

Fig. 2: Measurement results of a fluorescence spectrum.

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