Analysis Of Ki67 Staining
The tissue samples were visualized using a Leica microscope (Leica Microsystems, Wetzlar, Germany), and the Leica image manager software to enhance and photograph the sections. All sections were photographed and epidermal thickness was measured 50 places per section and the average epidermal thickness was calculated pr section (Fig. S5). The measurements were made on sections photographed at 10X enhancement. All sections were blinded to the observer during measurements.
Strengths:
Good method to measure the epidermal thickness as the keratinocytes are colored strongly blue and violet in contrast to stratum dermale and stratum corneum coloring pink. This makes it easy to measure exact thickness from stratum basale to stratum granulosum. …show more content…
Ki67 is a protein found in the nucleus of dividing cells and therefore necessary during cellular proliferation. The protein can be found in all phases of the cell cycle but not in resting cells. The Ki67 staining is used to determine growth of certain cell types and is therefore used on hyperproliferative cells such as tumor cells, but also in benign hyperproliferative diseases such as psoriasis. The staining reveals the proliferative active keratinocytes which can be found in stratum basale. Sept7-deficient mice receiving IMQ treatment (Fig S6. A) had very few keratinocytes in cellular divivion compared with IMQ treated mice (Fig S6. …show more content…
Very little of the specific gene is needed to amplify the gene. The method is also very specific as it combines the specificity of the primers and the fluorescence of the Taqman® probe.
Limitations: qPCR is an expensive method with high cost of equipment and cost of chemicals. The great sensitivity of this method makes is also its vulnerability, as the slightest error in the sample quality, RNA extraction or reverse transcriptation process can influence the final result. The result does not represent actual cytokine levels but only the amount of gene expression, as post-translational modifications of proteins are not accounted
Strengths:
Good method to measure the epidermal thickness as the keratinocytes are colored strongly blue and violet in contrast to stratum dermale and stratum corneum coloring pink. This makes it easy to measure exact thickness from stratum basale to stratum granulosum. …show more content…
Ki67 is a protein found in the nucleus of dividing cells and therefore necessary during cellular proliferation. The protein can be found in all phases of the cell cycle but not in resting cells. The Ki67 staining is used to determine growth of certain cell types and is therefore used on hyperproliferative cells such as tumor cells, but also in benign hyperproliferative diseases such as psoriasis. The staining reveals the proliferative active keratinocytes which can be found in stratum basale. Sept7-deficient mice receiving IMQ treatment (Fig S6. A) had very few keratinocytes in cellular divivion compared with IMQ treated mice (Fig S6. …show more content…
Very little of the specific gene is needed to amplify the gene. The method is also very specific as it combines the specificity of the primers and the fluorescence of the Taqman® probe.
Limitations: qPCR is an expensive method with high cost of equipment and cost of chemicals. The great sensitivity of this method makes is also its vulnerability, as the slightest error in the sample quality, RNA extraction or reverse transcriptation process can influence the final result. The result does not represent actual cytokine levels but only the amount of gene expression, as post-translational modifications of proteins are not accounted