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    Calibration of Microscopes

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
    Flag for inappropriate content
    27 views4 pages

    Calibration of Microscopes

    Uploaded by

    cezalyn
    Calibration of Microscopes

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
    Flag for inappropriate content
    of 4
    CALIBRATION OF MICROSCOPES FOR MAGNIFICATION AND RESOLUTION By A. W. Agar, Agar Scientific Limited, 66a Cambridge Road, Stansted, Bssex, CM24 SDA, UK. With the rapidly inereasing emphasis on quantitative results, itis very important to know what the magnification of a microscope really is. ‘The various available calibrations are described, together with their limitations, It is furthermore important to ensure that. a microscope is performing to specification, or at least to an adequate level for the experiment in hand. The appropriate tests for resolution and instrument performance are reviewed, Keywords: Mieroseope, Calibration, Magnification, Resolution henever we use any Kind of microscope, W be it a research light microscope, a scanning electron microscope ora TEM, ‘ye noe to know the magnification. The manuf turers hepllly provide this information in ap proximate form, and for many applications this is all ve noed to know. However, as sean as any work involving the dimensions ofthe specimen under examination is undertaken the instrument needs mave exact calibration, Calibrating the light microscope Measurements are normally made by using sgaticule within Uhe eyepiece, which les in the Fecal plane ofthe eye lens Hovevey, this grabcule isonly a comparative measure ast assumes the magnification ofthe objective lens and also the tbe length ofthe microscope which in some in struments can be varied, Its mecessary therfore to place a calibrated seale in the objet position, ad to pect the magnified image onto the plene the eyepiece gratiule, so thatthe latter may be calibrated. Sage mietometers may be either tramparent and covered with a cover gas ort: Nective and without a cover glass (for incident Tight instruments) {fig. 1). The ealibration is only valid fr the particular objective and eye lens beng wed and for the pertcular tube length at that time (ne assumes thatthe objective lens is fully served int its mount and Unt the eye lens is properly seated in the eyepiece tube. Once this alvation has been carried out for given pair of lenses, it will remain valid pro vided that there is nochange in any ofthe mech anieal parameters. Arecently develop test specimen is silieon chip which hasbeen electron beam waitten with ‘square mesh ones of approximetely 10 mito: retves, and with thicker lines every 500 micro | | Figure 1 meters was primarily designed forthe SBM but isfound very useful als for incident light mito mops, 28 the contrast i good (ig. 3). Calibrating the SEM "The electron lenses project a demagnifed image of the elatronsoutee on to the specimen under ‘examination, na seanning raster normally eon trolled by electromagnetic deflector coils and diven in synchronism with the display tube raster, The magnfiation is determined by the ‘Ste of the scanned raster on the display tube Aivided by the size ofthe rater onthe specimen. It is clear that the magnification therefore depends on the excitation ofthe scanning cols, ‘modified by any remanent magnetim o acide, tal eapacitance. tis quite possible thatthe two scan cnils wll nat yield exactly the same magni: eatin, so giving vise to orthogonal distortion. ‘The sean on the display tube is also subject to distortion and nonlinearity. Even after the stem iscalibeated,thore will bono guarentee that the calibration wil not ehange with time. ‘The magnification is alo rather sensitively de pendent on the working distance (objetve lens to specimen) whichis best determined in terms cf Ue objective lens curent. The calibration must take thi fetor into eecount. Since the magnification range of @ scanning letra mievazope starts within the range ofthe light nzroseope but goes upto very high values,

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