Whole mount

Ris, H. (1985). The cytoplasmic filament system in critical point-dried whole mounts and plastic-embedded section. J. Cell Biol. 100,1474-1487. 

Schmidt A., Naujoks-Manteuffel C. and Roth G. (1988). Olfactory and vomeronasal projections and the pathway of the Nervus terminalis in ten species of salamanders — a whole mount study employing the horseradish-peroxidase technique. Cell Tissue Res 251, 45-50. 

Woodson, K, J Gillespie, J Hanson, M Emmert-Buck, JM Phillips et al. 2004. Heterogeneous gene methyla-tion patterns among pre-invasive and cancerous lesions of the prostate a histopathologic study of whole mount prostate specimens. Prostate 60(1) 25-31. 

Fig. 20.1. Confocal images of whole mounts of the ovijector region of A suum stained with phalloidin-tetramethylrhodamine isothiocyanate (TRITC) to show muscle and with an anti-RFamide antiserum coupled to fluorescein isothiocyanate (FITC) to show FaRPergic nerves. (A) Main ventral nerve cord encircles opening of ovijector where it meets the body wall and is immunopositive for FaRPs. (B) Flat-fixed preparation of the ovijector showing circular muscles and tracts of parallel FaRPergic nerves (arrows). (C) Detail of the circular muscle of ovijector and associated nerves (arrows). (D) A FaRPergic cell body is localized in the ventral nerve cord at junction with ovijector and provides innervation to ovijector muscle.

Judice TN, Nelson NC, Beisel CL, Delimont DC, Fritzsch B, et al. 2002. Cochlear whole mount in situ hybridization identification of longitudinal and radial gradients. Brain Res Prot 9 65-76. 

Luque JM, Adams WB, NichoUs JG. 1998. Procedures for whole-mount immunohistochemistry and in situ hybridization of immature mammalian CNS. Brain Res Prot 2 165-173. 

Shifman MI, Selzer ME. 2000. In situ hybridization in whole-mounted lamprey spinal cord localization of netrin mRNA expression. J Neurosci Meth 104 19-25. 

Fluorescent Labeling of Surface or Intracellular Antigens in Whole-Mounts... 

The general immunocytochemical principles as applied to whole-mounts do not differ substantially from those applied to cells (see Chapters 16 and 17). However, several additional factors must be taken into account. Foremost is the problem of tissue penetration by reagents. Unlike sectioned material where intracellular components are directly exposed to the reagents, in whole-mount... 

Another example of how whole-mount technology has facilitated the study of three-dimensional arrays of filamentous networks is the visualization of cytoskeletal elements within the context of tissues. It is only through whole-... 

Fig. 1. Hydra oligactis whole mount labeled with monoclonal antibody JDl. (A) Isolated ganglionic neuron in the body column, scale bar = 50 om (B) hypostomal nerve net with sensory neurons of the mouth at the left and ganglionic neurons of the perihypostomal ring to the right, scale bar = 50 pm (C) cell bodies of hypostomal sensory neurons extending from the mesoglea (processes) to the surface of the ectoderm, scale bar = 25 pm.

It should be noted that an alternative approach that avoids some of the disadvantages of immunofluorescence is the use of enzyme-conjugated secondary antibodies (see Chapter 23). Although this approach sacrifices the resolution of a light-emitting source, low-power objectives compatible with thicker whole-mounts give optimal images (13). 

Fig. 2. Hypostome (mouth) of a Hydra oligactis whole mount labeled with monoclonal antibody DB5. (A) Nonconfocal image of sensory neurons and processes (B) confocal optical section of the same field as (A) illustrating details of neuronal cell bodies and processes. Scale bar = 25 pm.

Depending on the whole-mount specimen, fine forceps or needles can be nsed to arrange the specimen so that tissue is not folded prior to applying the top cover-slip. Two coverslips, rather than a slide and coverslip, are used so that both sides of the whole-mount may be examined. 

I would like to acknowledge Dr. Jack Dunne, who introduced me to monoclonal antibody technology and immunocytochemistry on whole-mounts and... 

Terada, M., Iwanaga, T., Takahasi-Iwanaga, H., Adachi, I., Arakawa, M., and Fujita, T. (1992) Calcitonin gene-related peptide (CGRP)-immunoreactive nerves in the tracheal epithelium of rats an immunohistochemical study by means of whole-mount preparations. Arc/t. Histol. Cytol. 55, 219-233. 

Liou, W. (1990) Whole-mount preparations of mouse lens epithelium for the fluorescent cytological study of actin. J. Microscopy 157, 239-245. 

Tam, P. K. and Boyd, G. P. (1990) Origin, conrse, and endings of abnormal enteric nerve fibres in Hirschspring s disease defined by whole-mount immnnohistochem-istry. J. Ped. Surg. 25, 457 61. 

Knratani, S., Tanaka, S., Ishikawa, Y., andZnkeran, C. (1988) Early development of the hypoglossal nerve in the chick embryo as observed by whole-mount nerve staining method. Am. J. Anal. 182,155-168. 

Plickert, G. and Kroiher, M. (1988) Proliferation kinetics and cell lineages can be stndied in whole-mounts and macerates by means of BrdU/anti-BrdU technique. Develop. 103, 791-794. 

Dent, J. A., Poison, A. G., and Klymkowsky, M. W. (1989) A whole-mount immnnocytochemical analysis of the expression of the intermediate filament protein vimentin in Xenopns. Develop. 105, 61-74. 

Gnstafsson, M. K. (1991) Skin the tapeworms before you stain their nervous system A new method for whole-mount immunocytochemistry. Parasitology Res. 77, 509-516. 

Fig. 3. Comparisons of wide-field (A) and confocal fluorescence images (B, mesoglea level C, apical) of rhodamine phalloidin-stained F-actin in a whole-mount hydra tentacle. The hydra was fixed and stained as described in Chapter 18. The bar represents 25 pm. All images were collected with a Nikon (New York) Microphot FX microscope (x40 objective lens). Confocal images were collected with the microscope connected to a Bio-Rad (Hercules, CA) MRC600 laser-scanning confocal system.

F. 3 Comparison of the neuromuscular junction pattern and muscular fiber organization in control and clofibrate-treated larvae, using whole-mount acetylcholinesterase staining. 

Fig. 8 Clofibrate impairs endotrophic lipid consumption in zebrafish larva (a-e) Whole-mount ORO staining of representative larvae are shown in lateral view with the anterior part to the left. Enlargement at the trunk level is shown in panels b and e. Control larva at 3 dpf (a) and 4 dpf (d, b). Clofibrate 0.75 mgA-treated larva at 3 dpf (c). Clofibrate 0.5 mg/l-treated larva at 4 dpf (f, e). e eye, da dorsal aorta, dlav dorsal longitudinal anastomotic vessel, h heart, i intestine, isv intersegmental vessel, pcv posterior cardinal vein, sb swim bladder, ys yolk sac. Reprinted from [24] with permission from Elsevier...

Fig. 2 Whole-mount T4 Immunofluorescence staining superposed on brightfield illumination of thyroid follicles in (a) 5 dpf control (0.1% DMSO used as a vehicle control), (b) 1.5 mM MMI-treated larvae, (c) 1 pM amiodarone-treated larvae, and (d) 50 nM T3-treated larvae. Heads of representative larvae are shown in ventral view with the anterior part on the left. Abbreviation e eye. Reprinted with permission from [46], Copyright 2009 American Chemical Society.

Fig. 3 Quantitative analysis of whole-mount T4-immunofluorescence signals for the Thyroxine Immunofluorescence Quantitative Disruption Test (TIQDT). The intrafollicular T4-content was calculated in the initial development of the TIQDT by measuring the average pixel intensity of the thyroid follicles. Reprinted with permission from [46],Copyright 2009 American Chemical Society.

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