Analysis Conditions (analysis + condition)

Distribution by Scientific Domains


Selected Abstracts


Influence of Sample Preparation, Staining Procedure and Analysis Conditions on Bull Sperm Head Morphometry using the Morphology Analyser Integrated Visual Optical System

REPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2001
A Boersma
The importance of standardizing the procedures of sample and slide preparation for computer-assisted morphologic analysis has been emphasized in human and veterinary andrology. The purpose of this study was to optimize slide preparation (dilution grade and sperm washing), staining procedures and analysis conditions (colour of light source and objective magnification) for the morphometric analysis of bull spermatozoa using the Hamilton Thorne morphology analyzer integrated visual optical system (IVOS). For experiment 1, one ejaculate was collected from one bull and diluted to 200 000,300 000 spermatozoa/,l. Slides were prepared and stained using seven different procedures: rapid Papanicolaou (PAP), rapid Papanicolaou with prolonged staining times (PAP+), Diff-Quik (DIF), haematoxylin (HEM), Farelly (FAR), Spermac (SPER) and the modified GZIN (MGZIN) staining. All slides were analysed using a Hamilton Thorne Morphology Analyser IVOS equipped alternatively with a red, green or blue light source, and a 40 or 100 oil immersion objective. Recognition and digitization errors as well as morphometric parameters were determined. The IVOS was unable to detect DIF-stained spermatozoa. The GZIN and the SPER staining as well as the blue light source led to unsatisfactory results. Among the staining methods examined, the FAR, HEM, PAP+, and PAP staining, preferably in combination with the green light source, and the 40 objective yielded optimal results concerning sperm recognition and digitization. The 100 objective did not allow reliable analysis of the sperm heads because of a frequently appearing digitization error. For experiment 2, three ejaculates were collected from each of three bulls and diluted to five dilution grades (100 000,500 000 spermatozoa/,l). An aliquot of each dilution grade was washed additionally. The percentage of correctly digitized sperm heads decreased with increasing spermatozoal concentration. However, the evaluation speed increased. The range of 200 000,300 000 spermatozoa/,l appeared to be a reasonable compromise for both criteria. Sperm washing failed to further improve the analysis results. Sperm head dimensions were influenced significantly by all variations of the methods in both experiments. In conclusion, using the proposed methods, the IVOS allows precise and reliable morphometric analyses of bull spermatozoa. The consistent application of these procedures may lead to an inter-laboratory standardization and to further establishment of generally accepted morphometric criteria used in human andrology (e.g. World Health Organisation or strict criteria). [source]


Rapid denaturing high-performance liquid chromatography (DHPLC) for mutation scanning of the transforming growth factor ,3 gene using a novel proof-reading polymerase

INTERNATIONAL JOURNAL OF IMMUNOGENETICS, Issue 5 2003
A. Bayat
Summary We have utilized a novel variation on the conventional denaturing high-performance liquid chromatography (DHPLC) technology, which we term rapid DHPLC, combining changes in instrumentation, cartridge technology and analysis conditions to enable significant increases in throughput to be achieved. In addition, the use of a novel proof-reading polymerase for sample amplification with a low misincorporation rate enables simplification of the DHPLC patterns and hence enhanced mutation detection recognition. This scheme for increasing DHPLC throughput has been tested by scanning the transforming growth factor (TGF) ,3 gene for the presence of mutations for which there is limited published or on-line data available regarding the presence of gene polymorphisms. TGF, isoforms have multiple roles in cell division, growth, proliferation, transformation and differentiation. TGF,3 is a TGF, cytokine isoform, and has an important role in embryogenesis, cell differentiation and wound healing. The TGF,3 gene consists of seven exons and six introns spanning 43 000 bp of the human genome on chromosome 14q23,24. The rapid DHPLC approach enabled scanning of all seven exons and part of the promoter region (1000 bp upstream from exon 1 in the 5,-flanking regions) of the TGF,3 gene in 95 Caucasian individuals in only 8 days, in comparison to the 17 days it would have previously taken. Mutations were clearly identified in the promoter region of the TGF,3 gene but were absent from the exonic regions. Understanding the genetic variations affecting the TGF,3 gene is important as this molecule has multiple regulatory functions on a variety of cell types. [source]


Isolation and potential existence of side population cells in adult human kidney

INTERNATIONAL JOURNAL OF UROLOGY, Issue 3 2008
Toshihiko Inowa
Abstract: The existence of adult stem-like cells such as side population (SP) cells is reported in various kinds of animal tissues, and we recently reported that mice kidney SP cells differentiate into multilineage. However, there has thus far been no report about human kidney SP cells. In the present study, we examined the existence of SP cells in human kidney tissue by using Hoechst 33342 staining and fluorescence-activated cell sorting analysis. We used porcine kidney tissue to optimize the analysis conditions for human tissue, and found that the SP population in human kidney was 1.3%. The existence of SP cells in human kidney suggests that the cells could be good targets for clinical renal regenerative medicine. [source]


Influence of Sample Preparation, Staining Procedure and Analysis Conditions on Bull Sperm Head Morphometry using the Morphology Analyser Integrated Visual Optical System

REPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2001
A Boersma
The importance of standardizing the procedures of sample and slide preparation for computer-assisted morphologic analysis has been emphasized in human and veterinary andrology. The purpose of this study was to optimize slide preparation (dilution grade and sperm washing), staining procedures and analysis conditions (colour of light source and objective magnification) for the morphometric analysis of bull spermatozoa using the Hamilton Thorne morphology analyzer integrated visual optical system (IVOS). For experiment 1, one ejaculate was collected from one bull and diluted to 200 000,300 000 spermatozoa/,l. Slides were prepared and stained using seven different procedures: rapid Papanicolaou (PAP), rapid Papanicolaou with prolonged staining times (PAP+), Diff-Quik (DIF), haematoxylin (HEM), Farelly (FAR), Spermac (SPER) and the modified GZIN (MGZIN) staining. All slides were analysed using a Hamilton Thorne Morphology Analyser IVOS equipped alternatively with a red, green or blue light source, and a 40 or 100 oil immersion objective. Recognition and digitization errors as well as morphometric parameters were determined. The IVOS was unable to detect DIF-stained spermatozoa. The GZIN and the SPER staining as well as the blue light source led to unsatisfactory results. Among the staining methods examined, the FAR, HEM, PAP+, and PAP staining, preferably in combination with the green light source, and the 40 objective yielded optimal results concerning sperm recognition and digitization. The 100 objective did not allow reliable analysis of the sperm heads because of a frequently appearing digitization error. For experiment 2, three ejaculates were collected from each of three bulls and diluted to five dilution grades (100 000,500 000 spermatozoa/,l). An aliquot of each dilution grade was washed additionally. The percentage of correctly digitized sperm heads decreased with increasing spermatozoal concentration. However, the evaluation speed increased. The range of 200 000,300 000 spermatozoa/,l appeared to be a reasonable compromise for both criteria. Sperm washing failed to further improve the analysis results. Sperm head dimensions were influenced significantly by all variations of the methods in both experiments. In conclusion, using the proposed methods, the IVOS allows precise and reliable morphometric analyses of bull spermatozoa. The consistent application of these procedures may lead to an inter-laboratory standardization and to further establishment of generally accepted morphometric criteria used in human andrology (e.g. World Health Organisation or strict criteria). [source]