Abstract
BackgroundStructured illumination microscopy has been extensively used in biological imaging due to its low cost and easy implementation. However, the lack of quantitative imaging capability limits its application in absolute irradiance measurements.MethodWe have developed a quantitative structured illumination microscopy image processing algorithm (QSIM) as a plugin for the widely used ImageJ software. QSIM can work with the raw images acquired by a traditional structured illumination microscope and can quantitatively measure photon numbers, with noise estimates for both wide-field images and sectioned images.Results and conclusionWe demonstrated the quantitative image processing capability of QSIM by imaging a mouse kidney section in 3D. The results show that QSIM can transform structured illumination microscopy from qualitative to quantitative, which is essential for demanding fluorescence imaging applications.Electronic supplementary materialThe online version of this article (doi:10.1186/1475-925X-14-4) contains supplementary material, which is available to authorized users.
Highlights
Structured illumination microscopy has been extensively used in biological imaging due to its low cost and easy implementation
The results show that quantitative structured illumination microscopy image processing algorithm (QSIM) can transform structured illumination microscopy from qualitative to quantitative, which is essential for demanding fluorescence imaging applications
The QSIM-calculated wide-field image, sectioned image, and their noise maps at a representative depth z = 4 μm are shown in Figure 5a-d, respectively
Summary
We demonstrated the quantitative image processing capability of QSIM by imaging a mouse kidney section in 3D.
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