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A simple and efficient method for preparation of isolated ovarian follicles for transmission electron microscopy

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Abstract

Objective

A simple method for preparation of isolated ovarian follicles for transmission electron microscopy (TEM) using transwell inserts is described.

Materials and methods

Pre-antral follicles were enzymatically isolated from mouse ovaries and cultured overnight on transwell insert polyester membranes. The following day, isolated ovarian follicles were processed for TEM by moving the transwell insert through successive wells containing the fixation and embedding reagents. After polymerization of the resin, the polyester membrane with the follicles embedded in the resin was disengaged from the transwell unit. The resin was sectioned. Semi-thin sections were stained with toluidine blue while ultra-thin sections were stained by uranyl acetate and examined by light microscopy and TEM, respectively.

Results

Isolated ovarian follicles were easily processed in groups for TEM. Follicles were well embedded and there appeared to be no loss of tissue during processing. The ultra-structure of processed isolated ovarian follicles was well preserved with little evidence of processing artifacts.

Conclusions

In situ processing and preparation of isolated ovarian follicles by first allowing attachment on transwell insert membranes was shown to be a simple, rapid and effective method for TEM.

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Authors and Affiliations

  1. Department of OB-GYN, Women’s Health Institute, Cleveland Clinic Foundation, Cleveland, OH, USA

    Nina Desai, Faten AbdelHafez, James Goldfarb & Tommaso Falcone

  2. Department of Imaging core, Cleveland Clinic Foundation, Cleveland, OH, USA

    Judith Drazba

  3. The Cleveland Clinic Fertility Center, 26900 Cedar Rd, Beachwood, OH, 44122, USA

    Nina Desai

Authors
  1. Nina Desai
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  2. Faten AbdelHafez
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  3. Judith Drazba
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  4. James Goldfarb
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  5. Tommaso Falcone
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Corresponding author

Correspondence to Nina Desai.

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Financial support: None

Capsule

Use of transwell insert membranes as a vehicle for the preparation of isolated ovarian follicles for TEM is a very simple and easy technique.

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Desai, N., AbdelHafez, F., Drazba, J. et al. A simple and efficient method for preparation of isolated ovarian follicles for transmission electron microscopy. J Assist Reprod Genet 27, 97–101 (2010). https://doi.org/10.1007/s10815-010-9389-4

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  • DOI: https://doi.org/10.1007/s10815-010-9389-4

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