Spinal Cord Immunofluorescence Imaging

After terminal anaesthesia by barbiturate overdose, mice were perfused transcardially with 4% paraformaldehyde and spinal cords processed for immunofluorescence as previously described^{60 (link),72 (link)}. Primary antibodies were: rat anti-Pecam1 (BD Biosciences 550274, 1:200). Secondary antibodies were: Alexa Fluor 555 Goat Anti Rat (1:200, Life Technologies, A21434). Immunofluorescence was imaged digitally on a slide scanner [Olympus VS-120 Slide scanner] or confocal microscope [Zeiss LSM880 + Airy fast module with ZEN 2 Black software (Zeiss, Oberkochen, Germany)]. Images were processed using ImageJ (NIH) or Imaris (Bitplane, version 9.0.0).

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Squair J.W., Gautier M., Kathe C., Anderson M.A., James N.D., Hutson T.H., Hudelle R., Qaiser T., Matson K.J., Barraud Q., Levine A.J., La Manno G., Skinnider M.A, & Courtine G. (2021). Confronting false discoveries in single-cell differential expression. Nature Communications, 12, 5692.

Publication 2021

rat anti-Pecam1 Alexa Fluor 555 Goat Anti Rat VS-120 Slide scanner LSM880 + Airy fast module ZEN 2 Black software

Airy Alexa fluor 555 Anaesthesia Antibodies Barbiturate Confocal microscope Goat Immunofluorescence Mice Overdose Paraformaldehyde Pecam1 Spinal cords

Corresponding Organization :

Other organizations : École Polytechnique Fédérale de Lausanne, International Collaboration On Repair Discoveries, University of British Columbia, National Institute of Neurological Disorders and Stroke, University of Lausanne

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Variable analysis

independent variables

Terminal anaesthesia by barbiturate overdose

dependent variables

Immunofluorescence of spinal cords

control variables

Transcardial perfusion with 4% paraformaldehyde
Spinal cord processing for immunofluorescence as previously described in references 60 and 72

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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