Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Morphological correlations with dimensional change during SEM specimen preparation

A Boyde, E Maconnachie

    Scanning Electron Microscopy
    |January 1, 1981
    PubMed
    Summary
    This summary is machine-generated.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    Topographically guided hierarchical mineralization.

    Materials today. Bio·2021
    Same author

    The plate-to-rod transition in trabecular bone loss is elusive.

    Royal Society open science·2021
    Same author

    Evaluation of laser ablation microtomy for correlative microscopy of hard tissues.

    Journal of microscopy·2018
    Same author

    The relationship between the edge of the chick blastoderm and the vitelline membrane.

    Wilhelm Roux' Archiv fur Entwicklungsmechanik der Organismen·2017
    Same author

    Multiscale alterations in bone matrix quality increased fragility in steroid induced osteoporosis.

    Bone·2015
    Same author

    Calcium silicate cement-induced remineralisation of totally demineralised dentine in comparison with glass ionomer cement: tetracycline labelling and two-photon fluorescence microscopy.

    Journal of microscopy·2014
    Same journal

    Statoconia formation in molluscan statocysts.

    Scanning electron microscopy·1986
    Same journal

    Statolith formation in Cnidaria: effects of cadmium on Aurelia statoliths.

    Scanning electron microscopy·1986
    Same journal

    Phase transitions of calcium oxalate trihydrate and epitaxy in the weddellite-whewellite system.

    Scanning electron microscopy·1986
    Same journal

    Experimental investigation of the genesis of struvite stones in cats.

    Scanning electron microscopy·1986
    Same journal

    Analysis of canine urinary stones using infrared spectroscopy and scanning electron microscopy.

    Scanning electron microscopy·1986
    Same journal

    Otoconia as test masses in biological accelerometers: what can we learn about their formation from evolutionary studies and from work in microgravity?

    Scanning electron microscopy·1986
    See all related articles

    Scanning electron microscopy (SEM) of soft tissues requires careful processing. Optimal results for mouse embryo limbs depend on fixation, dehydration, and drying methods to preserve morphology and minimize shrinkage.

    Area of Science:

    • Biological Sciences
    • Microscopy Techniques
    • Electron Microscopy

    Background:

    • Soft tissue processing for scanning electron microscopy (SEM) involves multiple steps.
    • Previous work detailed dimensional changes during SEM specimen preparation.
    • Mouse embryo limbs were utilized to investigate morphological outcomes.

    Purpose of the Study:

    • To assess the morphological findings associated with soft tissue processing for SEM.
    • To evaluate the effects of different fixation, dehydration, and drying methods.
    • To identify methods that minimize structural distortion and preserve specimen integrity.

    Main Methods:

    • Specimens were fixed using glutaraldehyde (GA) and/or osmium tetroxide (OsO4).
    • Drying methods included freeze-drying (FD) and critical point drying (CPD).

    Related Experiment Videos

  • Solvent evaporation drying (SED) was also investigated, with variations in atmospheric saturation.

    • Main Results:

    • Freeze-drying and critical point drying yielded acceptable SEM results for GA and GA+OsO4 fixed tissues.
    • Optimal volume retention in FD correlated with surface morphology, barring ice crystal damage.
    • Plasmalemma perforation in CPD of GA-fixed tissue was prevented by post-osmication or divalent cation treatment.
    • Solvent evaporation drying caused shrinkage, minimized by using saturated atmospheres of volatile solvents (e.g., Freon 113, diethyl ether).
    • Ethanol dehydration swelling was mitigated by starting with 70% or 100% ethanol.

    Conclusions:

    • Both freeze-drying and critical point drying are suitable for SEM of soft tissues, with careful control of fixation and freezing rates crucial for FD.
    • Critical point drying requires specific fixation protocols (post-osmication or cation treatment) to prevent plasmalemma damage.
    • Solvent evaporation drying can be optimized for minimal shrinkage using volatile solvents in saturated atmospheres.
    • Ethanol dehydration protocols can be adjusted to prevent swelling and sample disruption.