Immunofluorescence Imaging of Respiratory and Sperm Cells

Respiratory and sperm cell samples were treated and incubated with primary and secondary antibodies as reported in previous studies (Omran and Loges, 2009 (link); Cindrić et al., 2019 (link); Aprea et al., 2021a (link)). Antibodies and their applied dilutions are listed in Supplementary Table S2. Cell nuclei were stained using Hoechst33342 (Thermo Fischer Scientific, Waltham, United States). Samples were mounted in DAKO fluorescence mounting medium (Dako North America Inc., Carpinteria, United States). IF images were taken with a Zeiss AxioObserver Z1 Apotome (Carl Zeiss Meditec AG, Oberkochen, Germany) or Laser Scanning Microscope LSM 880 (Carl Zeiss Microscopy GmbH, Jena, Germany). Images were processed and exported using the ZEISS ZEN Imaging Software 2012 (Carl Zeiss Microscopy GmbH, Jena, Germany) and figure panels created with the Adobe Creative Suite CS5 (Adobe Systems, San José, United States).

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Aprea I., Wilken A., Krallmann C., Nöthe-Menchen T., Olbrich H., Loges N.T., Dougherty G.W., Bracht D., Brenker C., Kliesch S., Strünker T., Tüttelmann F., Raidt J, & Omran H. (2023). Pathogenic gene variants in CCDC39, CCDC40, RSPH1, RSPH9, HYDIN, and SPEF2 cause defects of sperm flagella composition and male infertility. Frontiers in Genetics, 14, 1117821.

Publication 2023

Hoechst33342 DAKO fluorescence mounting medium Zeiss AxioObserver Z1 Apotome Laser Scanning Microscope LSM 880

Antibodies Cell nuclei Dilutions Fluorescence Hoechst33342 Laser scanning microscope Microscopy Respiratory Sperm cell

Corresponding Organization :

Other organizations : University Hospital Münster, University of Münster

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Variable analysis

independent variables

Treatment of respiratory and sperm cell samples
Incubation with primary and secondary antibodies

dependent variables

Localization and/or expression of target proteins in respiratory and sperm cells

control variables

Cell nuclei staining using Hoechst33342
Mounting of samples in DAKO fluorescence mounting medium

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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