Login processing...

Trial ends in Request Full Access Tell Your Colleague About Jove
Click here for the English version


Protocole optimisé pour la rétine Wholemount Préparation pour l'imagerie et immunohistochimie

Published: December 13, 2013 doi: 10.3791/51018


Name Company Catalog Number Comments
Millicell Cell Culture Insert, 12 mm,
hydrophilic PTFE (Biopore), 0.4 μm
Millipore PICM01250
Insulin syringe, 1 ml Beckton Dickinson 309659
Scissors Fine Science Tools 15003-08 dissection
Forceps, Dumont #55, inox Fine Science Tools 11255-20 dissection
Cryostat Leica various
Confocal Microscope System Nikon various
Capillary glass World Precision Instruments 1B150F-4
P-97 Flaming/Brown Micropipette Puller Sutter Instrument Co.
Picospritzer III Parker Hannifin
Glass Bottom Culture Dishes MatTek Corporation P35G-0-14-C
Petri dish Falcon 1008
Disposable Graduated Transfer Pipettes VWR 16001-180
Multiwell plates, 24-well Beckton Dickinson 351147
Cover glass, #1 Electron Microscopy Sciences 72200-30
Polysine adhesion slides Electron Microscopy Sciences 63412-01
Microscope slides Globe 1301
Liquid Blocker Electron Microscopy Sciences 71312
VECTASHIELD mounting medium Vector Laboratories H-1000
OCT medium Sakura 4583
Parafilm laboratory film Fisher 13-374-10
Dow Corning high vacuum grease Sigma-Aldrich Z273554
Mouse anti-PSD95 Millipore MABN68 antibody
Donkey anti-mouse Alexa 568 Invitrogen A10037 antibody
Isolectin Alexa 488 Invitrogen I21411
Chemiblocker Chemicon 2170S
Triton X-100 Sigma-Aldrich T9284
Sulforhodamine Sigma-Aldrich 341738
Carbodiimide Thermo Scientific 22980 EDC
Paraformaldehyde Sigma-Aldrich P6148



  1. Csaba, G., Kovacs, P., Pallinger, E. EDAC fixation increases the demonstrability of biogenic amines in the unicellular Tetrahymena: a flow cytometric and confocal microscopic comparative analysis. Cell Biol. Int. 30, 345-348 (2006).
  2. Fletcher, E. L., Hack, I., Brandstatter, J. H., Wassle, H. Synaptic localization of NMDA receptor subunits in the rat retina. J. Comp. Neurol. 420, 98-112 (2000).
  3. Gastinger, M. J., O'Brien, J. J., Larsen, N. B., Marshak, D. W. Histamine immunoreactive axons in the macaque retina. Invest. Ophthalmol. Vis. Sci. 40, 487-495 (1999).
  4. Haverkamp, S., Wassle, H. Immunocytochemical analysis of the mouse retina. J. Comp. Neurol. 424, 1-23 (2000).
  5. Koulen, P., Fletcher, E. L., Craven, S. E., Bredt, D. S., Wassle, H. Immunocytochemical localization of the postsynaptic density protein PSD-95 in the mammalian retina. J. Neurosci. 18, 10136-10149 (1998).
  6. Pena, J. T., et al. miRNA in situ hybridization in formaldehyde and EDC-fixed tissues. Nat. Methods. 6, 139-141 (2009).
  7. Tymianski, M., et al. A novel use for a carbodiimide compound for the fixation of fluorescent and non-fluorescent calcium indicators in situ following physiological experiments. Cell Calcium. 21, 175-183 (1997).
  8. Sagdullaev, B. T., Eggers, E. D., Purgert, R., Lukasiewicz, P. D. Nonlinear interactions between excitatory and inhibitory retinal synapses control visual output. J. Neurosci. 31, 15102-15112 (2011).
  9. Toychiev, A. H., Sagdullaev, B., Yee, C. W., Ivanova, E., Sagdullaev, B. T. A time and cost efficient approach to functional and structural assessment of living neuronal tissue. J. Neurosci. Methods. 214, 105-112 (2013).
  10. Yee, C. W., Toychiev, A. H., Sagdullaev, B. T. Network deficiency exacerbates impairment in a mouse model of retinal degeneration. Front Syst. Neurosci. 6, 8 (2012).
  11. Ivanova, E., Muller, U., Wassle, H. Characterization of the glycinergic input to bipolar cells of the mouse retina. Eur. J. Neurosci. 23, 350-364 (2006).
  12. Dmitrieva, N. A., Strang, C. E., Keyser, K. T. Expression of alpha 7 nicotinic acetylcholine receptors by bipolar, amacrine, and ganglion cells of the rabbit retina. J. Histochem. Cytochem. 55, 461-476 (2007).
Protocole optimisé pour la rétine Wholemount Préparation pour l'imagerie et immunohistochimie
Play Video

Cite this Article

Ivanova, E., Toychiev, A. H., Yee,More

Ivanova, E., Toychiev, A. H., Yee, C. W., Sagdullaev, B. T. Optimized Protocol for Retinal Wholemount Preparation for Imaging and Immunohistochemistry. J. Vis. Exp. (82), e51018, doi:10.3791/51018 (2013).

Copy Citation Download Citation Reprints and Permissions
View Video

Get cutting-edge science videos from JoVE sent straight to your inbox every month.

Waiting X
Simple Hit Counter