nach oben

Erschienen in:

14.12.2017 | Methods Paper

Quantitative validation of immunofluorescence and lectin staining using reduced CLARITY acrylamide formulations

verfasst von: D. M. Krolewski, V. Kumar, B. Martin, R. Tomer, K. Deisseroth, R. M. Myers, A. F. Schatzberg, F. S. Lee, J. D. Barchas, W. E. Bunney, H. Akil, S. J. Watson Jr.

Erschienen in: Brain Structure and Function | Ausgabe 2/2018

Einloggen, um Zugang zu erhalten

Abstract

The CLARITY technique enables three-dimensional visualization of fluorescent-labeled biomolecules in clarified intact brain samples, affording a unique view of molecular neuroanatomy and neurocircuitry. It is therefore, essential to find the ideal combination for clearing tissue and detecting the fluorescent-labeled signal. This method requires the formation of a formaldehyde–acrylamide fixative-generated hydrogel mesh through which cellular lipid is removed with sodium dodecyl sulfate. Several laboratories have used differential acrylamide and detergent concentrations to achieve better tissue clearing and antibody penetration, but the potential effects upon fluorescent signal retention is largely unknown. In an effort to optimize CLARITY processing procedures we performed quantitative parvalbumin immunofluorescence and lectin-based vasculature staining using either 4 or 8% sodium dodecyl sulfate detergent in combination with different acrylamide formulas in mouse brain slices. Using both confocal and CLARITY-optimized lightsheet microscope-acquired images, we demonstrate that 2% acrylamide monomer combined with 0.0125% bis-acrylamide and cleared with 4% sodium dodecyl sulfate generally provides the most optimal signal visualization amongst various hydrogel monomer concentrations, lipid removal times, and detergent concentrations.

Nur mit Berechtigung zugänglich

Ando K, Laborde Q, Lazar A, Godefroy D, Youssef I, Amar M, Pooler A, Potier MC, Delatour B, Duyckaerts C (2014) Inside Alzheimer brain with CLARITY: senile plaques, neurofibrillary tangles and axons in 3-D. Acta Neuropathol 128(3):457–459. https://doi.org/10.1007/s00401-014-1322-y CrossRefPubMedPubMedCentral

Bastrup J, Larsen PH (2017) Optimized CLARITY technique detects reduced parvalbumin density in a genetic model of schizophrenia. J Neurosci Methods 283:23–32. https://doi.org/10.1016/j.jneumeth.2017.03.011 CrossRefPubMed

Chang EH, Argyelan M, Aggarwal M, Chandon TS, Karlsgodt KH, Mori S, Malhotra AK (2017) The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains. Neuroimage 147:253–261. https://doi.org/10.1016/j.neuroimage.2016.11.068 CrossRefPubMed

Chung K, Wallace J, Kim SY, Kalyanasundaram S, Andalman AS, Davidson TJ, Mirzabekov JJ, Zalocusky KA, Mattis J, Denisin AK, Pak S, Bernstein H, Ramakrishnan C, Grosenick L, Gradinaru V, Deisseroth K (2013) Structural and molecular interrogation of intact biological systems. Nature 497(7449):332–337. https://doi.org/10.1038/nature12107 CrossRefPubMedPubMedCentral

Costantini I, Ghobril JP, Di Giovanna AP, Allegra Mascaro AL, Silvestri L, Mullenbroich MC, Onofri L, Conti V, Vanzi F, Sacconi L, Guerrini R, Markram H, Iannello G, Pavone FS (2015) A versatile clearing agent for multi-modal brain imaging. Sci Rep 5:9808. https://doi.org/10.1038/srep09808 CrossRefPubMedPubMedCentral

DeFelipe J (1997) Types of neurons, synaptic connections and chemical characteristics of cells immunoreactive for calbindin-D28K, parvalbumin and calretinin in the neocortex. J Chem Neuroanat 14(1):1–19CrossRefPubMed

Epp JR, Niibori Y, Liz Hsiang HL, Mercaldo V, Deisseroth K, Josselyn SA, Frankland PW (2015) Optimization of CLARITY for Clearing Whole-Brain and Other Intact Organs(1,2,3). eNeuro. https://doi.org/10.1523/ENEURO.0022-15.2015 PubMedPubMedCentral

Gonzalez-Burgos G, Cho RY, Lewis DA (2015) Alterations in cortical network oscillations and parvalbumin neurons in schizophrenia. Biol Psychiatry 77(12):1031–1040. https://doi.org/10.1016/j.biopsych.2015.03.010 CrossRefPubMedPubMedCentral

Kawashima H, Sueyoshi S, Li H, Yamamoto K, Osawa T (1990) Carbohydrate binding specificities of several poly-N-acetyllactosamine-binding lectins. Glycoconj J 7(4):323–334CrossRefPubMed

Kim SY, Cho JH, Murray E, Bakh N, Choi H, Ohn K, Ruelas L, Hubbert A, McCue M, Vassallo SL, Keller PJ, Chung K (2015) Stochastic electrotransport selectively enhances the transport of highly electromobile molecules. Proc Natl Acad Sci USA 112(46):E6274–E6283. https://doi.org/10.1073/pnas.1510133112 CrossRefPubMedPubMedCentral

Lee H, Park JH, Seo I, Park SH, Kim S (2014) Improved application of the electrophoretic tissue clearing technology, CLARITY, to intact solid organs including brain, pancreas, liver, kidney, lung, and intestine. BMC Dev Biol 14:48. https://doi.org/10.1186/s12861-014-0048-3 CrossRefPubMedPubMedCentral

Lerner TN, Shilyansky C, Davidson TJ, Evans KE, Beier KT, Zalocusky KA, Crow AK, Malenka RC, Luo L, Tomer R, Deisseroth K (2015) Intact-brain analyses reveal distinct information carried by SNc dopamine subcircuits. Cell 162(3):635–647. https://doi.org/10.1016/j.cell.2015.07.014 CrossRefPubMedPubMedCentral

Lewis DA, Curley AA, Glausier JR, Volk DW (2012) Cortical parvalbumin interneurons and cognitive dysfunction in schizophrenia. Trends Neurosci 35(1):57–67. https://doi.org/10.1016/j.tins.2011.10.004 CrossRefPubMed

Liu AK, Hurry ME, Ng OT, DeFelice J, Lai HM, Pearce RK, Wong GT, Chang RC, Gentleman SM (2016) Bringing CLARITY to the human brain: visualization of Lewy pathology in three dimensions. Neuropathol Appl Neurobiol 42(6):573–587. https://doi.org/10.1111/nan.12293 CrossRefPubMed

Magliaro C, Callara AL, Mattei G, Morcinelli M, Viaggi C, Vaglini F, Ahluwalia A (2016) clarifying CLARITY: quantitative optimization of the diffusion based delipidation protocol for genetically labeled tissue. Front Neurosci 10:179. https://doi.org/10.3389/fnins.2016.00179 CrossRefPubMedPubMedCentral

Menegas W, Bergan JF, Ogawa SK, Isogai Y, Umadevi Venkataraju K, Osten P, Uchida N, Watabe-Uchida M (2015) Dopamine neurons projecting to the posterior striatum form an anatomically distinct subclass. Elife 4:e10032. https://doi.org/10.7554/eLife.10032 CrossRefPubMedPubMedCentral

Miller SJ, Rothstein JD (2016) Astroglia in Thick Tissue with Super Resolution and Cellular Reconstruction. PLoS One 11(8):e0160391. https://doi.org/10.1371/journal.pone.0160391 CrossRefPubMedPubMedCentral

Paxinos G, Franklin KBJ (2001) The mouse brain in stereotaxic coordinates. 2nd edn, Academic Press, San Diego

Phillips J, Laude A, Lightowlers R, Morris CM, Turnbull DM, Lax NZ (2016) Development of passive CLARITY and immunofluorescent labelling of multiple proteins in human cerebellum: understanding mechanisms of neurodegeneration in mitochondrial disease. Sci Rep 6:26013. https://doi.org/10.1038/srep26013 CrossRefPubMedPubMedCentral

Poguzhelskaya E, Artamonov D, Bolshakova A, Vlasova O, Bezprozvanny I (2014) Simplified method to perform CLARITY imaging. Mol Neurodegener 9:19. https://doi.org/10.1186/1750-1326-9-19 CrossRefPubMedPubMedCentral

Porter GA, Palade GE, Milici AJ (1990) Differential binding of the lectins Griffonia simplicifolia I and Lycopersicon esculentum to microvascular endothelium: organ-specific localization and partial glycoprotein characterization. Eur J Cell Biol 51(1):85–95PubMed

Renier N, Wu Z, Simon DJ, Yang J, Ariel P, Tessier-Lavigne M (2014) iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging. Cell 159(4):896–910. https://doi.org/10.1016/j.cell.2014.10.010 CrossRefPubMed

Richardson DS, Lichtman JW (2015) Clarifying tissue clearing. Cell 162(2):246–257. https://doi.org/10.1016/j.cell.2015.06.067 CrossRefPubMedPubMedCentral

Stefaniuk M, Gualda EJ, Pawlowska M, Legutko D, Matryba P, Koza P, Konopka W, Owczarek D, Wawrzyniak M, Loza-Alvarez P, Kaczmarek L (2016) Light-sheet microscopy imaging of a whole cleared rat brain with Thy1-GFP transgene. Sci Rep 6:28209. https://doi.org/10.1038/srep28209 CrossRefPubMedPubMedCentral

Susaki EA, Tainaka K, Perrin D, Yukinaga H, Kuno A, Ueda HR (2015) Advanced CUBIC protocols for whole-brain and whole-body clearing and imaging. Nat Protoc 10(11):1709–1727. https://doi.org/10.1038/nprot.2015.085 CrossRefPubMed

Tomer R, Ye L, Hsueh B, Deisseroth K (2014) Advanced CLARITY for rapid and high-resolution imaging of intact tissues. Nat Protoc 9(7):1682–1697. https://doi.org/10.1038/nprot.2014.123 CrossRefPubMedPubMedCentral

Treweek JB, Chan KY, Flytzanis NC, Yang B, Deverman BE, Greenbaum A, Lignell A, Xiao C, Cai L, Ladinsky MS, Bjorkman PJ, Fowlkes CC, Gradinaru V (2015) Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping. Nat Protoc 10(11):1860–1896. https://doi.org/10.1038/nprot.2015.122 CrossRefPubMedPubMedCentral

Woo J, Lee M, Seo JM, Park HS, Cho YE (2016) Optimization of the optical transparency of rodent tissues by modified PACT-based passive clearing. Exp Mol Med 48(12):e274. https://doi.org/10.1038/emm.2016.105 CrossRefPubMedPubMedCentral

Yang B, Treweek JB, Kulkarni RP, Deverman BE, Chen CK, Lubeck E, Shah S, Cai L, Gradinaru V (2014) Single-cell phenotyping within transparent intact tissue through whole-body clearing. Cell 158(4):945–958. https://doi.org/10.1016/j.cell.2014.07.017 CrossRefPubMedPubMedCentral

Ye L, Allen WE, Thompson KR, Tian Q, Hsueh B, Ramakrishnan C, Wang AC, Jennings JH, Adhikari A, Halpern CH, Witten IB, Barth AL, Luo L, McNab JA, Deisseroth K (2016) Wiring and molecular features of prefrontal ensembles representing distinct experiences. Cell 165(7):1776–1788. https://doi.org/10.1016/j.cell.2016.05.010 CrossRefPubMedPubMedCentral

Yu T, Qi Y, Zhu J, Xu J, Gong H, Luo Q, Zhu D (2017) Elevated-temperature-induced acceleration of PACT clearing process of mouse brain tissue. Sci Rep 7:38848. https://doi.org/10.1038/srep38848 CrossRefPubMedPubMedCentral

Zhang MD, Tortoriello G, Hsueh B, Tomer R, Ye L, Mitsios N, Borgius L, Grant G, Kiehn O, Watanabe M, Uhlen M, Mulder J, Deisseroth K, Harkany T, Hokfelt TG (2014) Neuronal calcium-binding proteins 1/2 localize to dorsal root ganglia and excitatory spinal neurons and are regulated by nerve injury. Proc Natl Acad Sci USA 111(12):E1149–E1158. https://doi.org/10.1073/pnas.1402318111 CrossRefPubMedPubMedCentral

Zheng H, Rinaman L (2016) Simplified CLARITY for visualizing immunofluorescence labeling in the developing rat brain. Brain Struct Funct 221(4):2375–2383. https://doi.org/10.1007/s00429-015-1020-0 CrossRefPubMed

Titel: Quantitative validation of immunofluorescence and lectin staining using reduced CLARITY acrylamide formulations
verfasst von: D. M. Krolewski
V. Kumar
B. Martin
R. Tomer
K. Deisseroth
R. M. Myers
A. F. Schatzberg
F. S. Lee
J. D. Barchas
W. E. Bunney
H. Akil
S. J. Watson Jr.
Publikationsdatum: 14.12.2017
Verlag: Springer Berlin Heidelberg
Erschienen in: Brain Structure and Function / Ausgabe 2/2018
Print ISSN: 1863-2653
Elektronische ISSN: 1863-2661
DOI: https://doi.org/10.1007/s00429-017-1583-z

Leitlinien kompakt für die Neurologie

Mit medbee Pocketcards sicher entscheiden.

^{Seit 2022 gehört die medbee GmbH zum Springer Medizin Verlag}

Kostenlos registrieren

Neu im Fachgebiet Neurologie

23.04.2024 | Demenz | Nachrichten

Update Neurologie

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.

Newsletter bestellen

Springer Medizin

Quantitative validation of immunofluorescence and lectin staining using reduced CLARITY acrylamide formulations

Abstract

Leitlinien kompakt für die Neurologie

Neu im Fachgebiet Neurologie

Demenzkranke durch Antipsychotika vielfach gefährdet

Parkinson-Therapie im Wandel – aktuelle Leitlinie im Fokus

Auf diese Krankheiten bei Geflüchteten sollten Sie vorbereitet sein

Hustenstiller lindert Agitation bei Alzheimer

Update Neurologie

Springer Medizin

Abstract

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 2/2018

Influence of attention and bolus volume on brain organization during swallowing

Cocaine increases dopaminergic connectivity in the nucleus accumbens

Analysis of the vasculature by immunohistochemistry in paraffin-embedded brains

Specific properties of the SI and SII somatosensory areas and their effects on motor control: a system neurophysiological study

Thalamic interactions of cerebellum and basal ganglia

Correction to: Dentate granule progenitor cell properties are rapidly altered soon after birth

Leitlinien kompakt für die Neurologie

Neu im Fachgebiet Neurologie

Demenzkranke durch Antipsychotika vielfach gefährdet

Parkinson-Therapie im Wandel – aktuelle Leitlinie im Fokus

Auf diese Krankheiten bei Geflüchteten sollten Sie vorbereitet sein

Hustenstiller lindert Agitation bei Alzheimer

Update Neurologie