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1

Flow cytometry in mammalian sperm sexing and quality measurement

100%
Bochenek M.
Biotechnologia
|
1998
|
issue 2
119-129
EN
Flow cytometry is a powerful technique for measurements of many structural and functional cell parameters. In sperm analyses it offers several advantages over the formerly used methods: high accuracy, possibility of measuring two or even three parameters simultaneously and analysis of a high number of cells per second. Moreover, at present it is the only technique which allows sperm sorting of X- and Y-fractions with high purity. The paper is a review of the flow cytometry applications in sperm sexing and measurements of quality parameters: membrane integrity, acrosome status, mitochondrial and esterase activity and sperm chromatin structure.
2

Flow cytometric estimation of ploidy in anisoploid sugar-beet populations

80%
Slowinska E., Steen P.
Journal of Applied Genetics
|
1995
|
vol. 36
|
issue 2
111-118
EN
Determination of ploidy level is a comon practice in sugar-beet breeding and seed production.Lately many laboratories have replaced microscopic chromosome counting by flow cytometric estimation of the nuclear DNA content. The possibility of direct estimation of cytological composition in anisoploid populations instead of individual plants analyses was investigated.Diploid, triploid and tetraploid sugar-beet lines as well as their mixtures were tested.The results obtained show that the suggested method is fast, simple and, after a proper transformation of the computer values, could be applied in breeding and seed production of polyploid crops.Precise estimation of contamination by small admixture of seeds or plants of undesirable ploidy, however, requires individual plant rather than population analyses.
3

Automated reticulocyte analysis ? new possibilities for haematology diagnostics

80%
Turowski D., Wysocka J.
Postępy Higieny i Medycyny Doświadczalnej
|
2000
|
vol. 54
|
issue 4
553-566
EN
This review describes a new method of reticulocyte analysis based on flow cytometry, better than manual reticulocyte counting. Reticulocyte percentage, absolut number, indexes of maturity (RMI- Reticulocyte Maturity Index, IRF ? Immature Reticulocyte Fraction) and others qualitative parameters express the erythropoietic activity of kidneys and bone marrow. The new proposed parameters can improve the diagnosis of anemias and others haematological disorders like leukemia, myelodysplastic syndrom. They also provide exactly and early information about the status of bone marrow after ablative chemotherapy and transplantation. Automated reticulocyte counting can create new potentials for haematology diagnosis.
4

Sperm sexing and semen quality measurement by flow cytometry ? state-of the-art

80%
Bochenek M.
|
2003
|
issue 1
106-115
EN
Flow cytometry is a modern and powerful technique for the measurements of many sperm cells? parameters as well as sperm sexing. In sperm examination, it offers several advantages over the formerly used methods: high accuracy, possibility of measurements two or even three parameters simultaneously and analysis of large number of cells per second. Moreover, at present, it is the only technique which allows reliable sperm sorting of X- and Y-fractions with high purity. The paper is a review of the flow cytometry applications in sperm sexing and measurements of sperm chromatin structure.
5

Karyological characterization of sugar beet gynogenetic lines cultured in vitro

61%
Svirshchevskaya A., Dolezel J.
Journal of Applied Genetics
|
2001
|
vol. 42
|
issue 1
21-32
EN
Flow cytometry was used to screen ploidy levels in 47 cultured in vitro sugar beet gynogenetic lines of various origin and age, obtained after plant regeneration from unfertilized ovules. When donor plants were diploid, the majority of regenerants were found to have cells with 1C, 2C and 4C relative DNA content (mainly haploid and diploid) and there were large differences in the rate of spontaneous in vitro chromosome doubling between individual homozygous lines. Six ovule-derived lines regenerated from fertile and sterile diploid donors of forty-five lines were solid diploids from the very early stages of their in vitro cultivation, and thus could not be classified as doubled haploids. In the case of tetraploid donor plants, the gynogenetic regenerants demonstrated 2x-ploidy level. The results obtained in chimeric plants with both haploid and diploid cells indicated the possibility to overcome mixoploidy by their re-cultivation through generative shoot tip culture. The flow cytometry method confirmed data obtained by conventional microscopic chromosome counting in dividing leaf cells and was found very useful for screening of a large number of regenerants and for characterizing the process of in vitro gynogenetic lines formation in sugar beet.
6

Phosphatidylserine externalization during bone marrow cell death after treatment of mice with WR-2721 and gamma-rays

61%
Mayur L., Bochenek M., Augustznek A.
Folia Biologica
|
2002
|
vol. 50
|
issue 1-2
95-100
EN
The cell surface exposure of phosphatidylserine (PS) and the plasma membrane impairment were assessed in the bone marrow of adult male Swiss mice exposed to a single 6 Gy dose of 60Co gamma-rays, and treated intraperitoneally with the aminothiol WR-2721 (Amifostine, S-2-/3-aminopropylamino/ethyl phosphorothioic acid), at a dose of 400 mg/kg body weight, 30 min prior to gamma-irradiation. The bone marrow cells were stained with a combination of fluoresceinated annexin V (annexin V - FITC) and propidium iodide (PI) at 3 h, 7 h, and 24 h after treatment of mice with WR-2721 and 60Co gamma-irradiation. The number of early apoptotic cells (annexin V - FITC positive/ PI negative), and late apoptotic and necrotic cells (annexin V - FITC positive/ PI positive), was increased at 3 h after exposure of mice to 60Co gamma-rays and thereafter declined with the frequency of apoptotic and necrotic cells remaining lower in WR-2721 pre-treated mice. Using the annexin V - FITC flow cytometric assay, the radioprotective effect of WR-2721 against induction of apoptosis and necrosis in normal cells of the haematopoietic system was shown.
7

Analysis of physiological state of single bacterial cell using fluorescent staining methods

61%
Sadowska J., Grajek W.
Biotechnologia
|
2009
|
issue 4
102-114
EN
The paper reviews fluorescent staining techniques allowing to diagnose the physiological state of bacterial cells. Different staining probes and a manner of their use for analysis of cell viability, membrane potential, membrane integrity, intracellular pH, respiratory activity, amount of nucleic acids, and activity of chosen intracellular enzymes are described. Range of examples of fluorescent staining for monitoring of physiological state of bacteria in natural environment and in biotechnological processes are presented.
8

Effect of exogenous opioid peptides on TNF--induced human neutrophil apoptosis in vitro

61%
Sulowska Z., Majewska E., Tchorzewski H., Klink M.
Archivum Immunologiae et Therapiae Experimentalis
|
2003
|
vol. 51
|
issue 4
267-272
EN
Polymorphonuclear leukocytes (neutrophils) apoptosis is an important mechanism regulating the life span and some functions of neutrophils at inflamed sites. The opioid peptides are present in the peripheral circulation and their concentrations rapidly increase as the result of stress and inflammation. The effect of opioid peptides such as met-enkephalin (M-ENK) and beta-endorphin (beta-END) on tumor necrosis factor alpha (TNF-alpha)-induced apoptosis in human neutrophils in vitro was investigated. Neutrophils isolated from peripheral blood were cultured in the absence or presence of 106-1010 M of opioid peptides for 8, 12 and 18 hours. Features of apoptotic neutrophils were measured by flow cytometric method based on analysis of apoptotic nuclei (DNA content). We found that M-ENK and beta-END enhanced both non-induced and TNF-alpha-induced neutrophil apoptosis in vitro in a dose-dependent manner. The effect of opioid peptides on modulation of neutrophil apoptosis was not reversed by opioid-receptor antagonist naloxone. The results suggest that M-ENK and beta-END can regulate neutrophil life span via apoptosis and in this way may participate in resolution of inflammation.
9

Instability of ploidy level during regeneration of transformed and non-transformed tobacco (Nicotiana tabacum L.)

61%
Sliwinska E., Zimny J., Drozdowska J.
|
2003
|
issue 3
260-266
EN
In plant tissue cultures, somaclonal variation is often observed. It can be an effect of the changes in the individual chromosome number or in the ploidy level. Flow cytometry, a fast and accurate method for the estimation of the nuclear DNA content, can be applied to study these changes. The DNA content in differentiated tissues of Nicotiana tabacum cultured in vitro was estimated using Partec CCA flow cytometer, starting from explant, through callus, up to regenerated shoots. The explant constituted stem segments of N. tabacum plants, non-transformed and transformed with gfp gene. Flow cytometric analysis showed differences in the proportion of 2C, 4C, 8C and 16C cells in plant tissue in different culture stages. Among the regenerated plantlets originated from non-transformed and transformed plants, diploid, tetraploid and mixoploid forms were observed. The transformation did not influence the share of cells representing different ploidy levels in the investigated plant material.
10

Analysis of peritoneal and head kidney phagocytes during Thioglycollate-elicited peritoneal inflammation in the goldfish, Carassius auratus

61%
Gruca P., Chadzinska M., Lackowska B., Plytycz B.
|
|
vol. 44
137-142
EN
Intraperitoneal injection of goldfish with Thioglycollate induced an acute peritoneal inflammation connected with an influx of leucocytes. The number and some properties (migratory activity, PMA-induced chemiluminescence) of leucocytes were analysed in parallel in suspensions of peritoneal and head kidney (HK) leucocytes. It was found that the increased number of peritoneal large granular cells corresponds with the decreased number of such leucocytes in HK. Moreover, the enhancement of migratory and chemiluminescent activity of peritoneal leucocytes corresponds with the inhibition of these activities in head kidney cells. The results strongly indicate that the head kidney, the major lymphomyeloid organ of teleosts, is the main source of leucocytes inhabiting the peritoneal cavity following induction of an acute inflammation.
11

Application of flow cytometry for the study of plant genomes

61%
Dolezel J.
|
|
issue 3
285-302
EN
During the last fifteen years, flow cytometry and sorting greatly contributed to the improvement of our knowledge of plant genome structure and function. This paper reviews the applications of flow cytometry for the analysis of isolated nuclei and chromosomes. Because of its speed, precision and convenience, this method of analysis of the nuclear DNA content finds an enormous number of applications which cover basic research, breeding and production. The results obtained with chromosome analysis and sorting indicate that the technique might greatly simplify the analysis of plant genomes at the molecular level.
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