Journal: Biomedical Optics Express

Article Title: Large-scale, cell-resolution volumetric mapping allows layer-specific investigation of human brain cytoarchitecture

doi: 10.1364/BOE.415555

Figure Lengend Snippet: (a) Schematic illustration of the SWITCH/TDE clearing method. (b) 1 mm-thick slice of an adult human brain sample before and after the treatment. (c) Images of SYTOX Green labeled tissue at different depths. Scale bar = 100μm. (d) Images of NeuN immunostained tissue at different depths. Scale bar = 50μm. Acquisition obtained with TPFM. (e) Representative images of cleared tissues immunostained with various antibodies and DAPI (4′,6-Diamidino-2-Phenylindole, Dihydrochloride). Scale bar = 50μm. Acronym list: Neuron-specific Nuclear protein (NeuN, all neurons), Microtubule-Associated Protein 2 (MAP2; pyramidal cells), Nonphosphorylated neurofilament protein (SMI32; pyramidal cells), Glutamic Acid Decarboxylase (GAD67; all GABAergic interneurons), Parvalbumin (PV; GABAergic interneurons subtype), Calbindin (CB; GABAergic interneurons subtype), Vasointestinal peptide (VIP; GABAergic interneurons subtype), Somatostatin (SST; GABAergic interneuron subtype), Neuropeptide Y (NPY; GABAergic interneuron subtype), Neurofilament (NF), Ionized calcium Binding Adaptor molecule 1 (Iba1; glial cells), Glial Fibrillary Acidic Protein (GFAP; glial cells), Glutamine synthetase (GluS), Vimentin (Vim; Microvasculature), Collagen IV (Coll IV; microvasculature).

Article Snippet: Sample pictures before and after the clearing process were acquired using a digital camera (Sony DSC-WX500), samples were kept soaked either in PBS or TDE. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Molecule Company Cat. n. Host P/M Dilution NeuN Merck ABN91 Chicken P 1:50 GAD67 Santa Cruz sc-28376 Mouse M 1:200 GAD65/67 St John’s Lab STJ93195 Rabbit P 1:200 PV Abcam ab11427 Rabbit P 1:200 PV Abcam ab32895 Goat P 1:200 CB Abcam ab207528 Rabbit M 1:200 VIP Abcam ab214244 Rabbit M 1:200 SST Abcam ab30788 Rat M 1:200 NPY Abcam ab6173 Sheep P 1:200 NPY Abcam ab112473 Mouse M 1:200 SMI-32 Merck NE1023 Mouse M 1:200 Neurofilament Abcam ab4680 Chicken P 1:200 GluS Merck MAB302 Mouse M 1:200 MAP2 Abcam ab5392 Chicken P 1:200 GFAP Abcam ab194324 Rabbit M 1:200 Iba1 Abcam ab195031 Rabbit M 1:200 Coll IV Abcam ab6586 Rabbit P 1:200 Vim Abcam ab8069 Mouse M 1:200 Anti-Rat IgG, AF 568 Abcam ab175475 Donkey P 1:200 Anti-Rabbit IgG, AF 568 Abcam ab175470 Donkey P 1:200 Anti-Chicken IgY, AF 568 Abcam ab175711 Goat P 1:200 Anti-Mouse IgG, AF 568 Abcam ab175700 Donkey P 1:200 Anti-Sheep IgG, AF 568 Abcam ab175712 Donkey P 1:200 Anti-Rabbit IgG, AF 488 Abcam ab150077 Goat P 1:200 Anti-Chicken IgY, AF 488 Abcam ab150169 Goat P 1:200 DAPI Thermo Fisher Scientific D1306 1:1000 SYTOX Green Thermo Fisher Scientific S7020 1:1000 Open in a separate window caption a8 Table summarizing the dyes tested in this study.

Techniques: Labeling, Binding Assay

Journal: Biomedical Optics Express

Article Title: Large-scale, cell-resolution volumetric mapping allows layer-specific investigation of human brain cytoarchitecture

doi: 10.1364/BOE.415555

Figure Lengend Snippet: Table summarizing the dyes tested in this study. The P/M column denotes polyclonal vs monoclonal antibodies. The same abbreviations used in are used. AF is a shorthand for Alexa Fluor.

Article Snippet: Sample pictures before and after the clearing process were acquired using a digital camera (Sony DSC-WX500), samples were kept soaked either in PBS or TDE. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Molecule Company Cat. n. Host P/M Dilution NeuN Merck ABN91 Chicken P 1:50 GAD67 Santa Cruz sc-28376 Mouse M 1:200 GAD65/67 St John’s Lab STJ93195 Rabbit P 1:200 PV Abcam ab11427 Rabbit P 1:200 PV Abcam ab32895 Goat P 1:200 CB Abcam ab207528 Rabbit M 1:200 VIP Abcam ab214244 Rabbit M 1:200 SST Abcam ab30788 Rat M 1:200 NPY Abcam ab6173 Sheep P 1:200 NPY Abcam ab112473 Mouse M 1:200 SMI-32 Merck NE1023 Mouse M 1:200 Neurofilament Abcam ab4680 Chicken P 1:200 GluS Merck MAB302 Mouse M 1:200 MAP2 Abcam ab5392 Chicken P 1:200 GFAP Abcam ab194324 Rabbit M 1:200 Iba1 Abcam ab195031 Rabbit M 1:200 Coll IV Abcam ab6586 Rabbit P 1:200 Vim Abcam ab8069 Mouse M 1:200 Anti-Rat IgG, AF 568 Abcam ab175475 Donkey P 1:200 Anti-Rabbit IgG, AF 568 Abcam ab175470 Donkey P 1:200 Anti-Chicken IgY, AF 568 Abcam ab175711 Goat P 1:200 Anti-Mouse IgG, AF 568 Abcam ab175700 Donkey P 1:200 Anti-Sheep IgG, AF 568 Abcam ab175712 Donkey P 1:200 Anti-Rabbit IgG, AF 488 Abcam ab150077 Goat P 1:200 Anti-Chicken IgY, AF 488 Abcam ab150169 Goat P 1:200 DAPI Thermo Fisher Scientific D1306 1:1000 SYTOX Green Thermo Fisher Scientific S7020 1:1000 Open in a separate window caption a8 Table summarizing the dyes tested in this study.

Techniques: Bioprocessing

Journal: Science Advances

Article Title: A causal coding variant regulating alternative splicing of DOC2A at 16p.11.2 GWAS locus influences susceptibility to schizophrenia

doi: 10.1126/sciadv.adw7667

Figure Lengend Snippet: ( A ) Representative traces of mEPSC in primary hippocampal neuron infected with control (empty vector), DOC2A Full-Length , or DOC2A ∆Val217-Pro218 AAV. ( B and C ) Quantification of mEPSC amplitude and frequency. N ≥ 29 neurons. ( D ) Representative traces of mIPSC in in primary hippocampal neuron infected with control, DOC2A Full-Length , or DOC2A ∆Val217-Pro218 AAV. ( E and F ) Quantification of mIPSC amplitude and frequency. N ≥ 20 neurons. ( G ) Representative gels of Western blot. ( H to K ) Quantification of synaptophysin, VAMP-2, GAD65, and GAD67 expression in hippocampus tissue overexpressing control, DOC2A Full-Length , or DOC2A ∆Val217-Pro218 . N = 3 in each group. Full Western blotting images are shown in fig. S16. Data are presented as the means ± SEM; * P < 0.05, ** P < 0.01, and *** P < 0.001. Error bars indicate the SEM.

Article Snippet: The primary antibodies were as follows: FLAG (Cell Signaling Technology, no. 8146S; 1:1000) and DOC2A (Invitrogen, no. PA5-31345; 1:1000), GAD67 (Proteintech, no. 10408-1-AP; 1:5000), GAD65 (Cell Signaling Technology, no. 5843T; 1:1000), synaptophysin (Sigma-Aldrich, no. S5768; 1:500), VAMP-2 (Synaptic Systems, no. 104211; 1:2000), α-tubulin (Proteintech, no. 66031-1-Ig; 1:20,000), and GAPDH (glyceraldehyde-3-phosphate dehydrogenase; Proteintech, no. 10494-1-AP; 1:20,000).

Techniques: Infection, Control, Plasmid Preparation, Western Blot, Expressing

Journal: eLife

Article Title: Ciliary neuropeptidergic signaling dynamically regulates excitatory synapses in postnatal neocortical pyramidal neurons

doi: 10.7554/eLife.65427

Figure Lengend Snippet:

Article Snippet: Antibody , αGAD67 (Goat polyclonal) , R and D Systems , Cat:AF2086, RRID: AB_2107724 , IF(1:2000).

Techniques: Transfection, Construct, Recombinant, Plasmid Preparation, shRNA, Software