Combined light microscope and scanning electron microscope, a new instrument for cell biology

Wouters, C. H.; Koerten, H. K.

doi:10.1016/0309-1651(82)90007-8

Cited by 19 publications

(9 citation statements)

References 3 publications

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“…T o investigate whether selective loss of certain cell types took place due to the preparation method, mononuclear cells were identified by the presence of non-specific cytoplasmic esterase activity, which is specific for monocytes. Staining for enzyme activity was performed on smears (Yam et al, 1971) or on cells in suspension (Nardiello et al, 1982).…”

Section: Incubation Of Cells For Esterase-activitymentioning

confidence: 99%

“…For LM a cytocentrifugation method (originally described by Leif et al, 1975) has been adapted for cell sedimentation in quantitative cytology (van Driel-Kulker et al, 1980). A well-defined and standardized cell preparation technique is equally important when quantitative L M DNA measurements are used in combination with SEM for the study of surface structures, using a recently developed instrument for combined SEM and LM (Ploem & Thaer, 1981;Wouters & Koerten, 1982). This microscope permits the simultaneous examination of cells with a scanning electron microscope and a light microscope and offers the advantages of a correlation between surface morphology and light microscopic cytochemical topology of cells.…”

Section: Introductionmentioning

confidence: 99%

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A centrifugation method for standardized sedimentation of mononuclear human blood cells on glass for scanning electron microscopy

Hesseling

et al. 1984

Journal of Microscopy

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SUMMARY A centrifugation method for depositing cells on cover‐glasses for scanning electron microscopy (SEM) is described. This centrifugation procedure provides a defined and standardized morphology of mononuclear cells from human blood. The method circumvents the highly variable flattening of some blood cells such as monocytes, observed with some methods. The SEM images show fine morphological surface details indicating a well‐preserved cell morphology. The cell recovery of the method is sufficiently high and the lymphocyte‐monocyte ratio in centrifugation preparations was found identical to the ratio in control smear preparations.

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Section: Incubation Of Cells For Esterase-activitymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A centrifugation method for standardized sedimentation of mononuclear human blood cells on glass for scanning electron microscopy

Hesseling

et al. 1984

Journal of Microscopy

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“…Early attempts to integrate a light microscope into an electron microscope date from the 1980s (Wouters & Koerten, 1982;Wouters, Koerten, Bonnet, Daems, & Ploem, 1986) and improved systems have been presented in recent years that are now commercially available (Agronskaia et al, 2008;Nishiyama et al, 2010;Zonnevylle et al, 2013). As CLEM has become more widely adopted, there has been a drive to improve the technique by integrating the two imaging modalities, thereby simplifying and expediting the correlative process.…”

Section: Introductionmentioning

confidence: 99%

Integrated Light and Scanning Electron Microscopy of GFP-Expressing Cells

Peddie¹,

Liv²,

Hoogenboom³

et al. 2014

Methods in Cell Biology

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“…In a system built by Wouters et al . in the 1980s (Wouters & Koerten, ; Wouters et al ., ; Wouters, ), the light microscope was integrated in a scanning electron microscope (SEM) under an angle of 45° with respect to the SEM optical axis. An illumination lens for transmission LM was added at the opposite side of the SEM objective lens.…”

Section: Introductionmentioning

confidence: 99%

Integration of a high‐NA light microscope in a scanning electron microscope

Zonnevylle

Tol

Liv

et al. 2013

Journal of Microscopy

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Summary We present an integrated light‐electron microscope in which an inverted high‐NA objective lens is positioned inside a scanning electron microscope (SEM). The SEM objective lens and the light objective lens have a common axis and focal plane, allowing high‐resolution optical microscopy and scanning electron microscopy on the same area of a sample simultaneously. Components for light illumination and detection can be mounted outside the vacuum, enabling flexibility in the construction of the light microscope. The light objective lens can be positioned underneath the SEM objective lens during operation for sub‐10 μm alignment of the fields of view of the light and electron microscopes. We demonstrate in situ epifluorescence microscopy in the SEM with a numerical aperture of 1.4 using vacuum‐compatible immersion oil. For a 40‐nm‐diameter fluorescent polymer nanoparticle, an intensity profile with a FWHM of 380 nm is measured whereas the SEM performance is uncompromised. The integrated instrument may offer new possibilities for correlative light and electron microscopy in the life sciences as well as in physics and chemistry.

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Combined light microscope and scanning electron microscope, a new instrument for cell biology

Cited by 19 publications

References 3 publications

A centrifugation method for standardized sedimentation of mononuclear human blood cells on glass for scanning electron microscopy

A centrifugation method for standardized sedimentation of mononuclear human blood cells on glass for scanning electron microscopy

Integrated Light and Scanning Electron Microscopy of GFP-Expressing Cells

Integration of a high‐NA light microscope in a scanning electron microscope

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