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The present study aimed to analyze different methods of mechanical isolation of buffalo preantral follicles (Experiment I), in vitro culture of isolated follicles in different groups of culture medium over collagen gel matrix (Experiment II) and subsequent in vitro development and survival of isolated preantral follicles (PFs) (Experiment III).
By mechanical isolation of the epicuticular 3D waxes, e.g., freezing in glycerol, the waxes can be removed from the cuticle, and transferred onto a smooth artificial substrate for microscopic investigations [36].
Mechanical isolation of the ICM was performed, and the ICM was plated onto feeders.
We used mechanical isolation of ICM from TE using tungsten needles.
Cellular Fractionation OSCs were resuspended in buffer I (10 mM HEPES, pH 7.9, 10 mM KCl, 0.1 mM MgCl2, 0.1mM EDTA, 0.1mM DTT, 0,1% NP40) and incubated on ice followed by mechanical isolation of nuclei with syringe and centrifugation.
After mechanical isolation of bundle sheath cells of C4 grasses, non-selective channels were formed with an exclusion limit of ca 1 kDa, consistent with open half-plasmodesmata [ 69, 70].
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Steps for mechanical isolation and culture of mesophyll cells are described in detail by Kärkönen et al. ([2011]).
Sympatric Parafontaria millipede species that lack effective precopulatory isolation were shown to have developed mechanical isolation by means of gonopod and cyphopod sexual morphological character displacement that effectively prevented interspecific transfer of the spermatophore to sympatric females [46].
Hence reproductive isolation between O. arachnitiformis and O. lupercalis growing in sympatry is usually achieved through a combination of (i) ethological isolation (the specific attraction of distinct pollinator taxa) and (ii) mechanical isolation (the differential attachment of pollen masses on the body of their respective pollinator).
In comparison, variation in floral size (PC1) could be more related to character displacement and the establishment of mechanical isolation between co-occurring species sharing a same functional group of pollinators [ 49, 50].
Certain floral traits may increase speciation rates through enhancement of mechanical isolation (Grant 1949, 1971, 1994 a ), where pollinators are restricted to a subset of visitors by the specialized fit of flowers to 'preferred' pollinators, or where flowers place pollen in a specific place on the pollinator (with stigma contact in the same location) not used by other species.
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