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Figure 1

From: Aqueous mounting media increasing tissue translucence improve image quality in Structured Illumination Microscopy of thick biological specimen

Figure 1

Quantitative comparison of raw SIM data quality. (A) MCNR calculated for Hodgkin’s lymphoma cells stained for the DNA with Hoechst 33258 and embedded in Vectashield. The red dashed line indicates an MCNR value threshold equal to 4 below which the data do not pass the evaluation and are not acceptable for SIM19. Each data-point represents a single cell measurement. Median and standard deviation values indicated in black. Below: example of MCNR maps and corresponding frequency spectra for raw SIM image sequences. Analogous data for Sorb70VE and H71VE media in (B) and (C), respectively. Low frequencies of high abundance are hidden behind a circular mask for clarity of data presentation. Note the strikingly different FT spectrum and MCNR map for Vectashield. Examples of FT spectra and MCNR maps for single cells in different media (images acquired with 1.518 immersion oil). All measurements were statistically different from respective measurements carried out in Vectashield, i.e. p-value in t-test <0.001. See Supplementary Fig. S4 for the same experiment for F80VE and S65VE media. Fluorescence signal dynamic range is compared in Supplementary Table 3.

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