Figure 6
From: Tissue clearing may alter emission and absorption properties of common fluorophores
The BABB-induced spectral alterations exacerbate crosstalk in multi-bandpass excitation scenarios in biological samples. BABB clearing exacerbates crosstalk in biological samples in the DAPI (a) and the 488 Filter range (b) in multi bandpass scenarios, which is shown by the reduced percentages of total DAPI and Alexa Fluor 488 signal intensities for BABB in the respective filter ranges. The stacked graphs illustrate the total fluorescent signal distribution of DAPI (blue), Alexa Fluor 488 (green), Alexa Fluor 568 (orange), and Alexa Fluor 647 (magenta) in PBS (left) and BABB (right) in biological samples (MEFs). Data are grouped by emission filter wavelength: in the DAPI filter range (a), the 488 nm filter range (b), the 568 nm filter range (c), and the 647 nm filter range (d). Samples were immunostained with an anti-GAPDH primary antibody, and respective secondary antibodies, or DAPI, and measured with a high content imaging system. Error bars indicate standard deviation. n = 8 independent wells and measurements for each datapoint.
