Ground state depletion microscopy as a tool for studying microglia–synapse interactions

• Published in: Journal of neuroscience research Vol. 99; no. 6; pp. 1515 - 1532
• Main Authors: Paasila, Patrick Jarmo, Fok, Sandra Y. Y., Flores‐Rodriguez, Neftali, Sajjan, Sujata, Svahn, Adam J., Dennis, Claude V., Holsinger, R. M. Damian, Kril, Jillian J., Becker, Thomas S., Banati, Richard B., Sutherland, Greg T., Graeber, Manuel B.
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01-06-2021; John Wiley and Sons Inc
• Subjects: Aged; Aged, 80 and over; Alzheimer disease; Alzheimer Disease - pathology; Alzheimer's disease; Animal tissues; Animals; Antibodies; Brain; Dementia disorders; Depletion; Electron microscopy; Female; Ground state; ground state depletion followed by individual molecule return microscopy; Humans; Larva; Larvae; Light diffraction; Light microscopy; Localization; Male; Mice; Microglia; Microglia - physiology; Microglia - ultrastructure; Microscopy; Microscopy - methods; Microscopy, Confocal; Middle Aged; Neurodegenerative diseases; Neuroimaging; Optical microscopy; Paraffin; Paraffins; post‐mortem archival human brain tissue; Presynaptic Terminals - physiology; Presynaptic Terminals - ultrastructure; RRID:AB_141874; RRID:AB_2199013; RRID:AB_2286948; RRID:AB_2534072; RRID:AB_2534076; RRID:AB_2535731; RRID:AB_2536183; RRID:AB_2633277; RRID:AB_324660; RRID:AB_839504; RRID:SCR_001622; RRID:SCR_002285; RRID:SCR_002798; RRID:SCR_013673; RRID:SCR_013726; super‐resolution microscopy; synapses; Synapses - physiology; Synapses - ultrastructure; Tissue Fixation; Zebrafish; RRID:AB_2286948; RRID:AB_2633277; RRID:SCR_002285; post-mortem archival human brain tissue; Alzheimer disease; ground state depletion followed by individual molecule return microscopy; RRID:AB_2199013; RRID:AB_2535731; RRID:SCR_013726; super-resolution microscopy; RRID:SCR_013673; RRID:AB_2534072; RRID:AB_324660; synapses; RRID:AB_2534076; RRID:AB_141874; zebrafish; RRID:SCR_001622; RRID:AB_2536183; RRID:SCR_002798; microglia; RRID:AB_839504
• ISSN: 0360-4012; 1097-4547
• DOI: 10.1002/jnr.24819

Ground state depletion followed by individual molecule return microscopy (GSDIM) has been used in the past to study the nanoscale distribution of protein co‐localization in living cells. We now demonstrate the successful application of GSDIM to archival human brain tissue sections including from Alzheimer's disease cases as well as experimental tissue samples from mouse and zebrafish larvae. Presynaptic terminals and microglia and their cell processes were visualized at a resolution beyond diffraction‐limited light microscopy, allowing clearer insights into their interactions in situ. The procedure described here offers time and cost savings compared to electron microscopy and opens the spectrum of molecular imaging using antibodies and super‐resolution microscopy to the analysis of routine formalin‐fixed paraffin sections of archival human brain. The investigation of microglia–synapse interactions in dementia will be of special interest in this context.