The influence of fixation on the morphology of mouse epidermis

Glasø, Morten; Hovig, Torstein

doi:10.1007/bf02899199

Cited by 10 publications

(5 citation statements)

References 27 publications

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“…Mechanical damage to tissue during the fixation process has been reported to cause intense staining in regions subjected to cutting or crushing (Cammermeyer 1972;Glasø and Hovig 1987). This is in stark contrast to our findings from the trout sacculus ( Fig.…”

Section: Fixation and Incidence Of Dhccontrasting

confidence: 99%

“…Likewise, dark cells were not observed in rat liver fixed in situ by vascular perfusion (Ganote and Moses 1968). In contrast, the mode of fixation (immersion or perfusion) did not seem to affect the number of dark cells in the skin of mice (Glasø and Hovig 1987). Our results ( Fig.…”

Section: Fixation and Incidence Of Dhccontrasting

confidence: 55%

“…In a study of nervous tissue from the brain, fixation by perfusion was found to minimise the post mortem traumatisation and reduce the number of dark neurones and pyknotic nuclei (Cammermeyer 1972). In contrast, the mode of fixation (immersion or perfusion) did not seem to affect the number of dark cells in the skin of mice (Glasø and Hovig 1987). In contrast, the mode of fixation (immersion or perfusion) did not seem to affect the number of dark cells in the skin of mice (Glasø and Hovig 1987).…”

Section: Fixation and Incidence Of Dhcmentioning

confidence: 97%

“…In mouse epidermis, the osmolality of the fixative was found to influence the number of dark cells (Ganote and Moses 1968;Glasø et al . 1986;Glasø and Hovig 1987;Glasø et al . 1989).…”

Section: Fixation and Incidence Of Dhcmentioning

confidence: 99%

“…Previously, the occurrence of dark cells in mammalian nervous tissue was suspected to be an artifact caused by mechanical maltreatment during the fixation process resulting in post-mortem traumatisation of cellular elements (Cammermeyer 1972). Likewise, dark cells in the skin of mice were reported to be induced by improper vehicle osmolarity (Glasø and Hovig 1987). Furthermore, the fixation method (immersion vs. perfusion) may also influence the incidence of dark cells, as found in a study of liver cells (Ganote and Moses 1968).…”

Section: Introductionmentioning

confidence: 99%

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Dark hair cells in the inner ear of the rainbow trout. A study of the influence of different fixation methods

Jensen

Jørgensen

2001

Acta Zoologica

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 Abstract Jensen, J.C. and Jørgensen, J.M. 2001. Dark hair cells in the inner ear of the rainbow trout. A study of the influence of different fixations methods. -Acta Zoologica (Stockholm) 82 : 79-88The occurrence of dark staining cells in different tissues has been suggested to be artefactual and caused during the fixation process. In inner ear sensory epithelia, dark hair cells (DHC) have been suggested to be apoptotic cells. We have examined whether dark cells represent dying cells or whether they are the results of fixation artefacts. The effects of buffer osmolarity and different fixation methods on the incidence of dark hair cells in the inner ear macula sacculi of the rainbow trout ( Oncorhynchus mykiss ) were investigated by light and electron microscopy. Glutaraldehyde in phosphate buffer with osmolarities of 0, 135, 225, 425, and 560 mosmol were used for fixation by immersion. For comparison, fixation by vascular perfusion as well as the effects of mechanical injury and delayed fixation were studied. DHC were found in all examined saccular maculae except for the delayed fixation protocol where almost all the sensory cells were lost. The number of DHC accounted for 2.5-12.9‰ of the sensory cells. Neither the buffer osmolarity nor the fixation method had significant effects on the frequencies of DHC. Mitotic cell division events were seen exclusively in the apical cell strata of the sensory epithelium. The DHC are suggested to be associated with apoptosis rather than fixation artefacts.

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Section: Fixation and Incidence Of Dhccontrasting

confidence: 99%

Section: Fixation and Incidence Of Dhccontrasting

confidence: 55%

Section: Fixation and Incidence Of Dhcmentioning

confidence: 97%

“…In mouse epidermis, the osmolality of the fixative was found to influence the number of dark cells (Ganote and Moses 1968;Glasø et al . 1986;Glasø and Hovig 1987;Glasø et al . 1989).…”

Section: Fixation and Incidence Of Dhcmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Dark hair cells in the inner ear of the rainbow trout. A study of the influence of different fixation methods

Jensen

Jørgensen

2001

Acta Zoologica

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Multivariate flow cytometry of epidermal regeneration provoked by a skin irritant and a tumor promoter

Kirkhus¹,

Glasø²,

Clausen³

1992

Cytometry

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The DNA content and the changes in cellular and nuclear size of isolated regenerating mouse epidermal basal cells were studied after topical application of the skin irritant cantharidin and the tumor promoter 12-0-tetradecanoylphorbol-13-acetate (TPA) to the back skin of hairless mice. The DNA and protein contents of isolated basal cells were stained with propidium iodide and fluorescein isothiocyanate, respectively, and analysed by flow cytometry using the total protein fluorescence as an estimate of cell size and the DNA fluorescence pulse width as an estimate of nuclear size. Transmission electron microscopy was used to identify cells sorted from regions in the bivariate DNNprotein distributions.The results showed that both chemicals induced an increase in cellular as well as nuclear size of the basal cells. The increase in size was higher in TPA treated than in cantharidin treated animals, and the bivariate DNNprotein distributions of TPA treated cells differed from those of cantharidin treated cells in that two subpopulations of cycling keratinocytes could be identified. These deviations are probably related to the higher proliferative response observed after TPA treatment and the possibility that proliferative subpopulations in epidermis respond differently to TPA. It may reflect mechanisms providing for a growth advantage of initiated cells, important in tumor promotion. About 8% of the cells in the suspensions from treated animals were non-cycling non-keratinocytes, probably infiltrating leukocytes.The results indicate a strong correlation between rapid regenerative cell cycle progression, i.e., reduced G1 transit time and increased cellular and nuclear size. The increase in cellular size was observed simultaneously in all cell cycle phases, well before the wave of cells entering S phase 16 h after application. This suggests that an increase in size may be essential for the subsequent initiation of DNA synthesis and the reduction in cell cycle time. The results support the hypothesis that the increase in cell size may be caused by gene activation or posttranscriptional processes rather than an increase due to an initial block in cell cycle progression.

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The influence of fixation on the relative amount of cytoplasmic ribosomes in mouse epidermal basal keratinocytes

Glasø

Iversen

Hovig

1988

Virchows Archiv B Cell Pathol

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The influence of fixation on the morphology of mouse epidermis

Cited by 10 publications

References 27 publications

Dark hair cells in the inner ear of the rainbow trout. A study of the influence of different fixation methods

Dark hair cells in the inner ear of the rainbow trout. A study of the influence of different fixation methods

Multivariate flow cytometry of epidermal regeneration provoked by a skin irritant and a tumor promoter

The influence of fixation on the relative amount of cytoplasmic ribosomes in mouse epidermal basal keratinocytes

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